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Sample records for generation active npc

  1. The mitochondrion interfering compound NPC-26 exerts potent anti-pancreatic cancer cell activity in vitro and in vivo.

    PubMed

    Dong, Yang-Yang; Zhuang, Yi-Huang; Cai, Wen-Jie; Liu, Yan; Zou, Wen-Bing

    2016-11-01

    The development of novel anti-pancreatic cancer agents is extremely important. Here, we investigated the anti-pancreatic cancer activity by NPC-26, a novel mitochondrion interfering compound. We showed that NPC-26 was anti-proliferative and cytotoxic to human pancreatic cancer cells, possibly via inducing caspase-9-dependent cell apoptosis. Pharmacological inhibition or shRNA-mediated silence of caspase-9 attenuated NPC-26-induced pancreatic cancer cell death and apoptosis. Further, NPC-26 treatment led to mitochondrial permeability transition pore (mPTP) opening in the cancer cells, which was evidenced by mitochondrial depolarization, ANT-1(adenine nucleotide translocator-1)-Cyp-D (cyclophilin-D) association and oxidative phosphorylation disturbance. mPTP blockers (cyclosporin and sanglifehrin A) or shRNA-mediated knockdown of key mPTP components (Cyp-D and ANT-1) dramatically attenuated NPC-26-induced pancreatic cancer cell apoptosis. Importantly, we showed that NPC-26, at a low concentration, potentiated gemcitabine-induced mPTP opening and subsequent pancreatic cancer cell apoptosis. In vivo, NPC-26 intraperitoneal injection significantly suppressed the growth of PANC-1 xenograft tumors in nude mice. Meanwhile, NPC-26 sensitized gemcitabine-mediated anti-pancreatic cancer activity in vivo. In summary, the results of this study suggest that NPC-26, alone or together with gemcitabine, potently inhibits pancreatic cancer cells possibly via disrupting mitochondrion.

  2. Acid sphingomyelinase activity is regulated by membrane lipids and facilitates cholesterol transfer by NPC2[S

    PubMed Central

    Oninla, Vincent O.; Breiden, Bernadette; Babalola, Jonathan O.; Sandhoff, Konrad

    2014-01-01

    During endocytosis, membrane components move to intraluminal vesicles of the endolysosomal compartment for digestion. At the late endosomes, cholesterol is sorted out mainly by two sterol-binding proteins, Niemann-Pick protein type C (NPC)1 and NPC2. To study the NPC2-mediated intervesicular cholesterol transfer, we developed a liposomal assay system. (Abdul-Hammed, M., B. Breiden, M. A. Adebayo, J. O. Babalola, G. Schwarzmann, and K. Sandhoff. 2010. Role of endosomal membrane lipids and NPC2 in cholesterol transfer and membrane fusion. J. Lipid Res. 51: 1747–1760.) Anionic lipids stimulate cholesterol transfer between liposomes while SM inhibits it, even in the presence of anionic bis(monoacylglycero)phosphate (BMP). Preincubation of vesicles containing SM with acid sphingomyelinase (ASM) (SM phosphodiesterase, EC 3.1.4.12) results in hydrolysis of SM to ceramide (Cer), which enhances cholesterol transfer. Besides SM, ASM also cleaves liposomal phosphatidylcholine. Anionic phospholipids derived from the plasma membrane (phosphatidylglycerol and phosphatidic acid) stimulate SM and phosphatidylcholine hydrolysis by ASM more effectively than BMP, which is generated during endocytosis. ASM-mediated hydrolysis of liposomal SM was also stimulated by incorporation of diacylglycerol (DAG), Cer, and free fatty acids into the liposomal membranes. Conversely, phosphatidylcholine hydrolysis was inhibited by incorporation of cholesterol, Cer, DAG, monoacylglycerol, and fatty acids. Our data suggest that SM degradation by ASM is required for physiological secretion of cholesterol from the late endosomal compartment, and is a key regulator of endolysosomal lipid digestion. PMID:25339683

  3. AAV9-NPC1 significantly ameliorates Purkinje cell death and behavioral abnormalities in mouse NPC disease.

    PubMed

    Xie, Chang; Gong, Xue-Min; Luo, Jie; Li, Bo-Liang; Song, Bao-Liang

    2017-03-01

    Niemann-Pick type C (NPC) disease is a fatal inherited neurodegenerative disorder caused by loss-of-function mutations in the NPC1 or NPC2 gene. There is no effective way to treat NPC disease. In this study, we used adeno-associated virus (AAV) serotype 9 (AAV9) to deliver a functional NPC1 gene systemically into NPC1(-/-) mice at postnatal day 4. One single AAV9-NPC1 injection resulted in robust NPC1 expression in various tissues, including brain, heart, and lung. Strikingly, AAV9-mediated NPC1 delivery significantly promoted Purkinje cell survival, restored locomotor activity and coordination, and increased the lifespan of NPC1(-/-) mice. Our work suggests that AAV-based gene therapy is a promising means to treat NPC disease.

  4. Plant phosphatidylcholine-hydrolyzing phospholipases C NPC3 and NPC4 with roles in root development and brassinolide signaling in Arabidopsis thaliana.

    PubMed

    Wimalasekera, Rinukshi; Pejchar, Premysl; Holk, André; Martinec, Jan; Scherer, Günther F E

    2010-05-01

    Phosphatidylcholine-hydrolyzing phospholipase C (PC-PLC) catalyzes the hydrolysis of phosphatidylcholine (PC) to generate phosphocholine and diacylglycerol (DAG). PC-PLC has a long tradition in animal signal transduction to generate DAG as a second messenger besides the classical phosphatidylinositol splitting phospholipase C (PI-PLC). Based on amino acid sequence similarity to bacterial PC-PLC, six putative PC-PLC genes (NPC1 to NPC6) were identified in the Arabidopsis genome. RT-PCR analysis revealed overlapping expression pattern of NPC genes in root, stem, leaf, flower, and silique. In auxin-treated P(NPC3):GUS and P(NPC4):GUS seedlings, strong increase of GUS activity was visible in roots, leaves, and shoots and, to a weaker extent, in brassinolide-treated (BL) seedlings. P(NPC4):GUS seedlings also responded to cytokinin with increased GUS activity in young leaves. Compared to wild-type, T-DNA insertional knockouts npc3 and npc4 showed shorter primary roots and lower lateral root density at low BL concentrations but increased lateral root densities in response to exogenous 0.05-1.0 μM BL. BL-induced expression of TCH4 and LRX2, which are involved in cell expansion, was impaired but not impaired in repression of CPD, a BL biosynthesis gene, in BL-treated npc3 and npc4. These observations suggest NPC3 and NPC4 are important in BL-mediated signaling in root growth. When treated with 0.1 μM BL, DAG accumulation was observed in tobacco BY-2 cell cultures labeled with fluorescent PC as early as 15 min after application. We hypothesize that at least one PC-PLC is a plant signaling enzyme in BL signal transduction and, as shown earlier, in elicitor signal transduction.

  5. c-Src activation promotes nasopharyngeal carcinoma metastasis by inducing the epithelial-mesenchymal transition via PI3K/Akt signaling pathway: a new and promising target for NPC

    PubMed Central

    Lu, Jinping; Xia, Weixiong; Yu, Yahui; Peng, Yongjian; Wang, Li; Wang, Gang; Ye, Yanfang; Yang, Jing; Liang, Hu; Kang, Tiebang; Lv, Xing

    2016-01-01

    Aberrant activation of cellular Src (c-Src), a non-receptor tyrosine kinase, could promote cancer progression through activating its downstream signaling pathways. However, the roles of c-Src and phosphorylated-Src (p-Src) in nasopharyngeal carcinoma (NPC) progression are rarely investigated. Herein, we have identified high c-Src concentrations in the serum of NPC patients with distant metastasis using high-throughput protein microarrays. Levels of c-Src in serum and p-Src in human primary NPC samples were unfavorable independent prognostic factors for cancer-specific survival, disease-free survival, and distant metastasis-free survival. Depletion or inactivation of c-Src in NPC cells using sgRNA with CRISPR/Cas9 system or PP2 decreased cell viability, colony formation, migration and invasion in vitro and metastasis in vivo. In contrast, these malignancies could be up-regulated by overexpressed c-Src in a NPC cell line with low-metastasis potential. Furthermore, p-Src was involved in promoting NPC cell metastasis by inducing the epithelial-mesenchymal transition (EMT) process via activating the PI3K/Akt pathway and cytoskeleton remodeling. The p-Src-induced EMT process could be retarded by PP2, which mediated by down-regulating the PI3K/Akt pathway. In conclusion, elevated levels of c-Src in serum and p-Src in primary NPC tissue correlated with poor outcomes of NPC patients. And aberrant activation of c-Src facilitated NPC cells with malignant potential, especially metastasis ability, which mediated by the PI3K/Akt pathway activation and sequentially induced the EMT process. These findings unveiled a promising approach for targeted therapy of advanced NPC. PMID:27078847

  6. Ezetimibe-sensitive cholesterol uptake by NPC1L1 protein does not require endocytosis

    PubMed Central

    Johnson, Tory A.; Pfeffer, Suzanne R.

    2016-01-01

    Human NPC1L1 protein mediates cholesterol absorption in the intestine and liver and is the target of the drug ezetimibe, which is used to treat hypercholesterolemia. Previous studies concluded that NPC1L1-GFP protein trafficking is regulated by cholesterol binding and that ezetimibe blocks NPC1L1-GFP function by inhibiting its endocytosis. We used cell surface biotinylation to monitor NPC1L1-GFP endocytosis and show that ezetimibe does not alter the rate of NPC1L1-GFP endocytosis in cultured rat hepatocytes grown under normal growth conditions. As expected, NPC1L1-GFP endocytosis depends in part on C-terminal, cytoplasmically oriented sequences, but endocytosis does not require cholesterol binding to NPC1L1’s N-terminal domain. In addition, two small- molecule inhibitors of general (and NPC1L1-GFP) endocytosis failed to inhibit the ezetimibe-sensitive uptake of [3H]cholesterol from taurocholate micelles. These experiments demonstrate that cholesterol uptake by NPC1L1 does not require endocytosis; moreover, ezetimibe interferes with NPC1L1’s cholesterol adsorption activity without blocking NPC1L1 internalization in RH7777 cells. PMID:27075173

  7. Drosophila melanogaster NPC2 Proteins Bind Bacterial Cell Wall Components and May Function in Immune Signal Pathways

    PubMed Central

    Shi, Xiu-Zhen; Zhong, Xue; Yu, Xiao-Qiang

    2012-01-01

    ML (MD-2 (myeloid differentiation factor 2)-related Lipid-recognition) is a conserved domain identified in MD-2, MD-1, NPC2 (Niemann-Pick disease type C2), and mite major allergen protein from animals, plants, and fungi. Vertebrate members of the ML family proteins, such as NPC2 and MD-2, play important roles in lipid metabolism and immune signaling pathway. MD-2 is an essential co-receptor in the lipopolysaccharide (LPS)/Toll-like receptor 4 (TLR4) signaling pathway. Insects contain multiple ML genes, arbitrarily named md-2- or npc2-like genes. However, whether insect ML genes have functions similar to vertebrate md-2 is unknown. In Drosophila melanogaster, there are eight npc2 genes (npc2a-h), and they can be further divided into three subgroups based on the numbers of cysteine residues (6, 7 and 8 Cys) in the mature proteins. The purpose of this study is to investigate whether any Drosophila npc2 genes may have functions in immune signaling pathways. We chose npc2a, npc2e and npc2h genes representing the three subgroups for this study. We showed that recombinant NPC2a, NPC2e and NPC2h not only bound to LPS and lipid A, but also bound to peptidoglycan (PG) and lipoteichoic acid (LTA), a property that has not been reported previously for vertebrate NPC2 or MD-2. More importantly, we showed that over-expression of NPC2a and NPC2e activated diptericin promoter reporter in S2 cells stimulated by PG, suggesting that NPC2e and NPC2a may play a role in the immune deficiency (Imd) pathway. This is the first in vitro study about NPC2 proteins in innate immunity of D. melanogaster. PMID:22580186

  8. FTY720/fingolimod increases NPC1 and NPC2 expression and reduces cholesterol and sphingolipid accumulation in Niemann-Pick type C mutant fibroblasts.

    PubMed

    Newton, Jason; Hait, Nitai C; Maceyka, Michael; Colaco, Alexandria; Maczis, Melissa; Wassif, Christopher A; Cougnoux, Antony; Porter, Forbes D; Milstien, Sheldon; Platt, Nicholas; Platt, Frances M; Spiegel, Sarah

    2017-04-01

    Niemann-Pick type C (NPC) disease is a fatal neurodegenerative disorder caused by mutations in NPC1 or NPC2 with decreased functions leading to lysosomal accumulation of cholesterol and sphingolipids. FTY720/fingolimod, used for treatment of multiple sclerosis, is phosphorylated by nuclear sphingosine kinase 2, and its active phosphorylated form (FTY720-P) is an inhibitor of class I histone deacetylases. In this study, administration of clinically relevant doses of FTY720 to mice increased expression of NPC1 and -2 in brain and liver and decreased cholesterol in an SphK2-dependent manner. FTY720 greatly increased expression of NPC1 and -2 in human NPC1 mutant fibroblasts that correlated with formation of FTY720-P and significantly reduced the accumulation of cholesterol and glycosphingolipids. In agreement with this finding, FTY720 pretreatment of human NPC1 mutant fibroblasts restored transport of the cholera toxin B subunit, which binds ganglioside GM1, to the Golgi apparatus. Together, these findings suggest that FTY720 administration can ameliorate cholesterol and sphingolipid storage and trafficking defects in NPC1 mutant fibroblasts. Because neurodegeneration is the main clinical feature of NPC disease, and FTY720 accumulates in the CNS and has several advantages over available histone deacetylase inhibitors now in clinical trials, our work provides a potential opportunity for treatment of this incurable disease.-Newton, J., Hait, N. C., Maceyka, M., Colaco, A., Maczis, M., Wassif, C. A., Cougnoux, A., Porter, F. D., Milstien, S., Platt, N., Platt, F. M., Spiegel, S. FTY720/fingolimod increases NPC1 and NPC2 expression and reduces cholesterol and sphingolipid accumulation in Niemann-Pick type C mutant fibroblasts.

  9. Analysis and carrier-based modulation of Z-source NPC inverters

    NASA Astrophysics Data System (ADS)

    Dehghan, Seyed Mohammad; Mohamadian, Mustafa; Gharekhani, Reza

    2012-08-01

    Multi-level neutral-point-clamped (NPC) inverters are widely used in high and medium voltage applications. However, these inverters are only buck-type converters. Recently z-source NPC inverters have been proposed as the buck-boost three-level inverter. New inverters use a z-source network as the input stage. This article analyses the features and limitations of two main topologies of z-source NPC inverters (dual z-source network NPC inverter and single z-source network NPC inverter). The low-frequency ripple of the inductor current and the capacitor voltage of the z-source networks in the z-source NPC inverters are surveyed. This article also proposes two novel PWM methods for the z-source NPC inverters, which can be easily implemented experimentally. In the proposed methods, generated switching signals for the conventional NPC inverter are modified for the z-source NPC inverter using a simple logic circuit considering the boost factors and the power sharing ratio. The performance of the proposed simple PWM methods is verified by simulation and experimental results.

  10. Transfer of cholesterol by the NPC team.

    PubMed

    Vance, Jean E

    2010-08-04

    The mechanisms of intracellular cholesterol transport are largely unknown. In this issue of Cell Metabolism, Wang et al. (2010) identify amino acid residues on the lumenal lysosomal protein Niemann-Pick C2 (NPC2) that are required for intralysosomal transfer of endocytosed cholesterol to membrane-bound NPC1 via a process that avoids movement of hydrophobic cholesterol through the aqueous phase.

  11. NPC1, intracellular cholesterol trafficking and atherosclerosis.

    PubMed

    Yu, Xiao-Hua; Jiang, Na; Yao, Ping-Bo; Zheng, Xi-Long; Cayabyab, Francisco S; Tang, Chao-Ke

    2014-02-15

    Post-lysosomal cholesterol trafficking is an important, but poorly understood process that is essential to maintain lipid homeostasis. Niemann-Pick type C1 (NPC1), an integral membrane protein on the limiting membrane of late endosome/lysosome (LE/LY), is known to accept cholesterol from NPC2 and then mediate cholesterol transport from LE/LY to endoplasmic reticulum (ER) and plasma membrane in a vesicle- or oxysterol-binding protein (OSBP)-related protein 5 (ORP5)-dependent manner. Mutations in the NPC1 gene can be found in the majority of NPC patients, who accumulate massive amounts of cholesterol and other lipids in the LE/LY due to a defect in intracellular lipid trafficking. Liver X receptor (LXR) is the major positive regulator of NPC1 expression. Atherosclerosis is the pathological basis of coronary heart disease, one of the major causes of death worldwide. NPC1 has been shown to play a critical role in the atherosclerotic progression. In this review, we have summarized the role of NPC1 in regulating intracellular cholesterol trafficking and atherosclerosis.

  12. NPC2 — EDRN Public Portal

    Cancer.gov

    NPC2, a secreted protein containing a lipid recognition domain, may function in regulating the transport of cholesterol through the late endosomal/lysosomal system. NPC2 may be involved in the regulation of the lipid composition of sperm membranes during the maturation in the epididymis. Mutations in this gene have been associated with Niemann-Pick disease, type C2 and frontal lobe atrophy.

  13. A Single Residue in Ebola Virus Receptor NPC1 Influences Cellular Host Range in Reptiles

    PubMed Central

    Ndungo, Esther; Herbert, Andrew S.; Raaben, Matthijs; Obernosterer, Gregor; Biswas, Rohan; Miller, Emily Happy; Wirchnianski, Ariel S.; Carette, Jan E.; Brummelkamp, Thijn R.; Whelan, Sean P.

    2016-01-01

    ABSTRACT Filoviruses are the causative agents of an increasing number of disease outbreaks in human populations, including the current unprecedented Ebola virus disease (EVD) outbreak in western Africa. One obstacle to controlling these epidemics is our poor understanding of the host range of filoviruses and their natural reservoirs. Here, we investigated the role of the intracellular filovirus receptor, Niemann-Pick C1 (NPC1) as a molecular determinant of Ebola virus (EBOV) host range at the cellular level. Whereas human cells can be infected by EBOV, a cell line derived from a Russell’s viper (Daboia russellii) (VH-2) is resistant to infection in an NPC1-dependent manner. We found that VH-2 cells are resistant to EBOV infection because the Russell’s viper NPC1 ortholog bound poorly to the EBOV spike glycoprotein (GP). Analysis of panels of viper-human NPC1 chimeras and point mutants allowed us to identify a single amino acid residue in NPC1, at position 503, that bidirectionally influenced both its binding to EBOV GP and its viral receptor activity in cells. Significantly, this single residue change perturbed neither NPC1’s endosomal localization nor its housekeeping role in cellular cholesterol trafficking. Together with other recent work, these findings identify sequences in NPC1 that are important for viral receptor activity by virtue of their direct interaction with EBOV GP and suggest that they may influence filovirus host range in nature. Broader surveys of NPC1 orthologs from vertebrates may delineate additional sequence polymorphisms in this gene that control susceptibility to filovirus infection. IMPORTANCE Identifying cellular factors that determine susceptibility to infection can help us understand how Ebola virus is transmitted. We asked if the EBOV receptor Niemann-Pick C1 (NPC1) could explain why reptiles are resistant to EBOV infection. We demonstrate that cells derived from the Russell’s viper are not susceptible to infection because EBOV

  14. Glycosylation inhibition reduces cholesterol accumulation in NPC1 protein-deficient cells.

    PubMed

    Li, Jian; Deffieu, Maika S; Lee, Peter L; Saha, Piyali; Pfeffer, Suzanne R

    2015-12-01

    Lysosomes are lined with a glycocalyx that protects the limiting membrane from the action of degradative enzymes. We tested the hypothesis that Niemann-Pick type C 1 (NPC1) protein aids the transfer of low density lipoprotein-derived cholesterol across this glycocalyx. A prediction of this model is that cells will be less dependent upon NPC1 if their glycocalyx is decreased in density. Lysosome cholesterol content was significantly lower after treatment of NPC1-deficient human fibroblasts with benzyl-2-acetamido-2-deoxy-α-D-galactopyranoside, an inhibitor of O-linked glycosylation. Direct biochemical measurement of cholesterol showed that lysosomes purified from NPC1-deficient fibroblasts contained at least 30% less cholesterol when O-linked glycosylation was blocked. As an independent means to modify protein glycosylation, we used Chinese hamster ovary ldl-D cells defective in UDP-Gal/UDP-GalNAc 4-epimerase in which N- and O-linked glycosylation can be controlled. CRISPR generated, NPC1-deficient ldl-D cells supplemented with galactose accumulated more cholesterol than those in which sugar addition was blocked. In the absence of galactose supplementation, NPC1-deficient ldl-D cells also transported more cholesterol from lysosomes to the endoplasmic reticulum, as monitored by an increase in cholesteryl [(14)C]-oleate levels. These experiments support a model in which NPC1 protein functions to transfer cholesterol past a lysosomal glycocalyx.

  15. Clues to the mechanism of cholesterol transfer from the structure of NPC1 middle lumenal domain bound to NPC2

    PubMed Central

    Li, Xiaochun; Saha, Piyali; Li, Jian; Blobel, Günter; Pfeffer, Suzanne R.

    2016-01-01

    Export of LDL-derived cholesterol from lysosomes requires the cooperation of the integral membrane protein Niemann–Pick C1 (NPC1) and a soluble protein, Niemann–Pick C2 (NPC2). Mutations in the genes encoding these proteins lead to Niemann–Pick disease type C (NPC). NPC2 binds to NPC1’s second (middle), lumenally oriented domain (MLD) and transfers cholesterol to NPC1’s N-terminal domain (NTD). Here, we report the 2.4-Å resolution crystal structure of a complex of human NPC1–MLD and NPC2 bearing bound cholesterol-3-O-sulfate. NPC1–MLD uses two protruding loops to bind NPC2, analogous to its interaction with the primed Ebola virus glycoprotein. Docking of the NPC1–NPC2 complex onto the full-length NPC1 structure reveals a direct cholesterol transfer tunnel between NPC2 and NTD cholesterol binding pockets, supporting the “hydrophobic hand-off” cholesterol transfer model. PMID:27551080

  16. N-terminal domain of the cholesterol transporter Niemann-Pick C1-like 1 (NPC1L1) is essential for α-tocopherol transport.

    PubMed

    Kamishikiryo, Jun; Haraguchi, Misaki; Nakashima, Shunsuke; Tasaka, Yuka; Narahara, Hiroe; Sugihara, Narumi; Nakamura, Tetsuya; Morita, Tetsuo

    2017-04-29

    Both cholesterol and α-tocopherol are essential lipophilic nutrients for humans and animals. Although cholesterol in excess causes severe problems such as coronary heart disease, it is a necessary component of cell membranes and is the precursor for the biosynthesis of steroid hormones and bile acids. Niemann-Pick C1-like 1 (NPC1L1) is a cholesterol transporter that is highly expressed in the small intestine and liver in humans and plays an important role in cholesterol homeostasis. Cholesterol promotes NPC1L1 endocytosis, which is an early step in cholesterol uptake. Furthermore, α-tocopherol is the most active form of vitamin E, and sufficient amounts of vitamin E are critical for health. It has been reported that NPC1L1 mediates α-tocopherol absorption; however, the mechanisms underlying this process are unknown. In this study, we found that treatment of cells that stably express NPC1L1-GFP with α-tocopherol promotes NPC1L1 endocytosis, and the NPC1L1 inhibitor, ezetimibe, efficiently prevents the α-tocopherol-induced endocytosis of NPC1L1. Cholesterol binding to the N-terminal domain (NTD) of NPC1L1 (NPC1L1-NTD) is essential for NPC1L1-mediated cholesterol absorption. We found that α-tocopherol competitively binds NPC1L1-NTD with cholesterol. Furthermore, when cells stably expressed NPC1L1ΔNTD-GFP, α-tocopherol could not induce the endocytosis of NPC1L1ΔNTD. Taken together, these results demonstrate that NPC1L1 recognizes α-tocopherol via its NTD and mediates α-tocopherol uptake through the same mechanism as cholesterol absorption.

  17. Active Nitrogen Generation.

    DTIC Science & Technology

    1987-06-01

    Project Officer FOR THE COMMANDER GERALD A. HASEN HARROACKERMANN Major, USAF Lt Col, USAF Chief, Advanced Chemical Laser Br Chief, Laser Science...SUPPLEMENTARY NOTATION 17. COSATI CODES I. SUJIECT TERMS lCntilnale o nnreqre if neetmay and identiy by Nocb awnubr) FlO GROUP Sue. n. Chemical lasers ...evalua- tion of the H + NF2 flame as a potential N2(A) generator for a high energy chemical laser . llAccesii For NTIS CRA&I l V IC TAR_

  18. RNA Export through the NPC in Eukaryotes.

    PubMed

    Okamura, Masumi; Inose, Haruko; Masuda, Seiji

    2015-03-20

    In eukaryotic cells, RNAs are transcribed in the nucleus and exported to the cytoplasm through the nuclear pore complex. The RNA molecules that are exported from the nucleus into the cytoplasm include messenger RNAs (mRNAs), ribosomal RNAs (rRNAs), transfer RNAs (tRNAs), small nuclear RNAs (snRNAs), micro RNAs (miRNAs), and viral mRNAs. Each RNA is transported by a specific nuclear export receptor. It is believed that most of the mRNAs are exported by Nxf1 (Mex67 in yeast), whereas rRNAs, snRNAs, and a certain subset of mRNAs are exported in a Crm1/Xpo1-dependent manner. tRNAs and miRNAs are exported by Xpot and Xpo5. However, multiple export receptors are involved in the export of some RNAs, such as 60S ribosomal subunit. In addition to these export receptors, some adapter proteins are required to export RNAs. The RNA export system of eukaryotic cells is also used by several types of RNA virus that depend on the machineries of the host cell in the nucleus for replication of their genome, therefore this review describes the RNA export system of two representative viruses. We also discuss the NPC anchoring-dependent mRNA export factors that directly recruit specific genes to the NPC.

  19. Active droplet generation in microfluidics.

    PubMed

    Chong, Zhuang Zhi; Tan, Say Hwa; Gañán-Calvo, Alfonso M; Tor, Shu Beng; Loh, Ngiap Hiang; Nguyen, Nam-Trung

    2016-01-07

    The reliable generation of micron-sized droplets is an important process for various applications in droplet-based microfluidics. The generated droplets work as a self-contained reaction platform in droplet-based lab-on-a-chip systems. With the maturity of this platform technology, sophisticated and delicate control of the droplet generation process is needed to address increasingly complex applications. This review presents the state of the art of active droplet generation concepts, which are categorized according to the nature of the induced energy. At the liquid/liquid interface, an energy imbalance leads to instability and droplet breakup.

  20. Simulations of Active Vortex Generators

    NASA Technical Reports Server (NTRS)

    Mansour, N. N.; Koumoutsakos, P.; Merriam, Marshal (Technical Monitor)

    1996-01-01

    We are interested in the study, via numerical simulations, of active vortex generators. Vortex generators may be used to modify the inner part of the boundary layer or to control separation thus enhancing the performance and maneuverability of aerodynamic configurations. We consider generators that consist of a surface cavity elongated in the streamwise direction and partially covered with a moving lid that at rest lies flush with the boundary. Streamwise voracity is generated and ejected due to the oscillatory motion of the lid. The present simulations c Implement relevant experimental investigations of active vortex generators that have been conducted at NASA Ames Research Center and Stanford University. Jacobson and Reynolds used a piezoelectric device in water, allowing for small amplitude high frequency oscillations. They placed the lid asymmetrically on the cavity and observed a strong outward velocity at the small gap of the cavity. Saddoughi used a larger mechanically driven device in air to investigate this flow and observed a jet emerging from the wide gap of the configuration, contrary to the findings of Jacobson and Reynolds. More recently, Lachowiez and Wlezien are investigating the flow generated by an electro-mechanically driven lid to be used for assertion control in aerodynamic applications. We are simulating the flows generated by these devices and we are conducting a parametric study that would help us elucidate the physical mechanisms present in the flow. Conventional computational schemes encounter difficulties when simulating flows around complex configurations undergoing arbitrary motions. Here we present a formulation that achieves this task on a purely Lagrangian frame by extending the formulation presented by Koumoutsakos, Leonard and Pepin. The viscous effects are taken into account by modifying the strength of the particles, whereas fast multipole schemes employing hundreds of thousands ol'particle's allow for high resolution simulations

  1. Simulations of Active Vortex Generators

    NASA Technical Reports Server (NTRS)

    Mansour, N. N.; Koumoutsakos, P.; Merriam, Marshal (Technical Monitor)

    1996-01-01

    We are interested in the study, via numerical simulations, of active vortex generators. Vortex generators may be used to modify the inner part of the boundary layer or to control separation thus enhancing the performance and maneuverability of aerodynamic configurations. we consider generators that consist of a surface cavity elongated in the streamwise direction and partially covered with a moving lid that at rest lies flush with the boundary. Streamwise vorticity is generated and ejected due to the oscillatory motion of the lid. The present simulations complement relevant experimental investigations of active vortex generators that have been conducted at NASA Ames Research Center and Stanford University used a piezoelectric device in water, allowing for small amplitude high frequency oscillations. They placed the lid asymmetrically on the cavity and observed a strong outward velocity at the small gap of the cavity. Saddoughi used a larger mechanically driven device in air to investigate this flow and observed a jet emerging from the wide gap of the configuration, contrary to the findings of Jacobson and Reynolds We are simulating the flows generated by these devices and we are conducting a parametric study that would help us elucidate the physical mechanisms present in the flow. Conventional computational schemes encounter difficulties when simulating flows around complex configurations undergoing arbitrary motions. Here we present a formulation that achieves this task on a purely Lagrangian frame by extending the formulation presented by Koumoutsakos, Leonard and Pepin (1994). The viscous effects are taken into account by modifying the strength Of the particles, whereas fast multipole schemes employing hundreds of thousands of particles allow for high resolution simulations. We shall present simulation results of an oscillating plate at various Reynolds numbers and Strouhal frequencies.

  2. NPC1 late endosomes contain elevated levels of non-esterified (‘free’) fatty acids and an abnormally glycosylated form of the NPC2 protein

    PubMed Central

    2005-01-01

    NPC (Niemann–Pick type C) disease is a rare lipidosis characterized by the accumulation of LDL (low-density lipoprotein)-derived non-esterified cholesterol in the E/L (endosomal/lysosomal) system. The gene products that are responsible for the two NPC complementation groups are distinct and dissimilar, yet their cellular and disease phenotypes are virtually indistinguishable. To investigate the relationship between NPC1 and NPC2 and their potential role in NPC disease pathogenesis, we have developed a method for the rapid and efficient isolation of late endocytic vesicles from mouse liver by magnetic chromatography. Late endosomes from Wt (wild-type) and NPC1 mice were found to differ not only in their cholesterol and sphingomyelin content, as expected, but also in their non-esterified (‘free’) fatty acid content, with NPC1 vesicles showing an approx. 7-fold increase in non-esterified fatty acid levels compared with Wt vesicles. Furthermore, we show that the NPC2 protein is in an incompletely deglycosylated form in NPC1 late endosomes by a mechanism that is specific to the NPC2 protein and not a global aberration of protein glycosylation/deglycosylation or trafficking, since NPC2 secreted from NPC1 cells is indistinguishable from that secreted from Wt cells. Also, a greater proportion of the normally soluble cellular NPC2 protein partitions with detergent-insoluble late endosomal internal membrane domains in NPC1 vesicles. In addition, we show that, although a small amount of the NPC2 protein associates with these membranes in Wt vesicles, this localization becomes much more pronounced in NPC1 vesicles. These results suggest that the function of the NPC2 protein may be compromised as well in NPC1 endosomes, which might explain the paradoxical phenotypic similarities of the two NPC disease complementation groups. PMID:15896196

  3. Transcriptomic Biomarkers for Tuberculosis: Evaluation of DOCK9. EPHA4, and NPC2 mRNA Expression in Peripheral Blood

    PubMed Central

    de Araujo, Leonardo S.; Vaas, Lea A. I.; Ribeiro-Alves, Marcelo; Geffers, Robert; Mello, Fernanda C. Q.; de Almeida, Alexandre S.; Moreira, Adriana da S. R.; Kritski, Afrânio L.; Lapa e Silva, José R.; Moraes, Milton O.; Pessler, Frank; Saad, Maria H. F.

    2016-01-01

    Lately, much effort has been made to find mRNA biomarkers for tuberculosis (TB) disease/infection with microarray-based approaches. In a pilot investigation, through RNA sequencing technology, we observed a prominent modulation of DOCK9, EPHA4, and NPC2 mRNA abundance in the blood of TB patients. To corroborate these findings, independent validations were performed in cohorts from different areas. Gene expression levels in blood were evaluated by quantitative real-time PCR (Brazil, n = 129) or reanalysis of public microarray data (UK: n = 96; South Africa: n = 51; Germany: n = 26; and UK/France: n = 63). In the Brazilian cohort, significant modulation of all target-genes was observed comparing TB vs. healthy recent close TB contacts (rCt). With a 92% specificity, NPC2 mRNA high expression (NPC2high) showed the highest sensitivity (85%, 95% CI 65%–96%; area under the ROC curve [AUROC] = 0.88), followed by EPHA4 (53%, 95% CI 33%–73%, AUROC = 0.73) and DOCK9 (19%, 95% CI 7%–40%; AUROC = 0.66). All the other reanalyzed cohorts corroborated the potential of NPC2high as a biomarker for TB (sensitivity: 82–100%; specificity: 94–97%). An NPC2high profile was also observed in 60% (29/48) of the tuberculin skin test positive rCt, and additional follow-up evaluation revealed changes in the expression levels of NPC2 during the different stages of Mycobacterium tuberculosis infection, suggesting that further studies are needed to evaluate modulation of this gene during latent TB and/or progression to active disease. Considering its high specificity, our data indicate, for the first time, that NPC2high might serve as an accurate single-gene biomarker for TB. PMID:27826286

  4. Nanoparticulate carriers (NPC) for oral pharmaceutics and nutraceutics.

    PubMed

    Lopes, C M; Martins-Lopes, P; Souto, E B

    2010-02-01

    The introduction of nanoparticulate carriers (NPC) in the pharmaceutic and nutraceutic fields has changed the definitions of disease management and treatment, diagnosis, as well as the supply food chain in the agri-food sector. NPC composed of synthetic polymers, proteins or polysaccharides gather interesting properties to be used for oral administration of pharmaceutics and nutraceutics. Oral administration remains the most convenient way of delivering drugs (e.g. peptides, proteins and nucleic acids) since these suffer similar metabolic pathways as food supply. Recent advances in biotechnology have produced highly potent new molecules however with low oral bioavailability. A suitable and promising approach to overcome their sensitivity to chemical and enzymatic hydrolysis as well as the poor cellular uptake, would be their entrapment within suitable gastrointestinal (GI) resistant NPC. Increasing attention has been paid to the potential use of NPC for peptides, proteins, antioxidants (carotenoids, omega fatty acids, coenzyme Q10), vitamins, probiotics, for oral administration. This review focuses on the most important materials to produce NPC for oral administration, and the most recent achievements in the production techniques and bioactives successfully delivered by these means.

  5. Miglustat Reverts the Impairment of Synaptic Plasticity in a Mouse Model of NPC Disease

    PubMed Central

    D'Arcangelo, G.; Grossi, D.; Racaniello, M.; Cardinale, A.; Zaratti, A.; Rufini, S.; Cutarelli, A.; Tancredi, V.; Merlo, D.; Frank, C.

    2016-01-01

    Niemann-Pick type C disease is an autosomal recessive storage disorder, characterized by abnormal sequestration of unesterified cholesterol within the late endolysosomal compartment of cells and accumulation of gangliosides and other sphingolipids. Progressive neurological deterioration and insurgence of symptoms like ataxia, seizure, and cognitive decline until severe dementia are pathognomonic features of the disease. Here, we studied synaptic plasticity phenomena and evaluated ERKs activation in the hippocampus of BALB/c NPC1−/− mice, a well described animal model of the disease. Our results demonstrated an impairment of both induction and maintenance of long term synaptic potentiation in NPC1−/− mouse slices, associated with the lack of ERKs phosphorylation. We then investigated the effects of Miglustat, a recent approved drug for the treatment of NPCD. We found that in vivo Miglustat administration in NPC1−/− mice was able to rescue synaptic plasticity deficits, to restore ERKs activation and to counteract hyperexcitability. Overall, these data indicate that Miglustat may be effective for treating the neurological deficits associated with NPCD, such as seizures and dementia. PMID:26885401

  6. EhNPC1 and EhNPC2 Proteins Participate in Trafficking of Exogenous Cholesterol in Entamoeba histolytica Trophozoites: Relevance for Phagocytosis

    PubMed Central

    Bolaños, Jeni; Betanzos, Abigail; Javier-Reyna, Rosario; García- Rivera, Guillermina; Huerta, Miriam; Pais-Morales, Jonnatan; González-Robles, Arturo; Rodríguez, Mario A.

    2016-01-01

    Entamoeba histolytica, the highly phagocytic protozoan causative of human amoebiasis lacks the machinery to synthesize cholesterol. Here, we investigated the presence of NPC1 and NPC2 proteins in this parasite, which are involved in cholesterol trafficking in mammals. Bioinformatics analysis revealed one Ehnpc1 and two Ehnpc2 genes. EhNPC1 appeared as a transmembrane protein and both EhNPC2 as peripheral membrane proteins. Molecular docking predicted that EhNPC1 and EhNPC2 bind cholesterol and interact with each other. Genes and proteins were identified in trophozoites. Serum pulse-chase and confocal microscopy assays unveiled that after trophozoites sensed the cholesterol source, EhNPC1 and EhNPC2 were organized around the plasma membrane in a punctuated pattern. Vesicles emerged and increased in number and size and some appeared full of cholesterol with EhNPC1 or EhNPC2 facing the extracellular space. Both proteins, but mostly EhNPC2, were found out of the cell associated with cholesterol. EhNPC1 and cholesterol formed networks from the plasma membrane to the nucleus. EhNPC2 appeared in erythrocytes that were being ingested by trophozoites, co-localizing with cholesterol of erythrocytes, whereas EhNPC1 surrounded the phagocytic cup. EhNPC1 and EhNPC2 co-localized with EhSERCA in the endoplasmic reticulum and with lysobisphosphatidic acid and EhADH (an Alix protein) in phagolysosomes. Immunoprecipitation assays confirmed the EhNPC1 and EhNPC2 association with cholesterol, EhRab7A and EhADH. Serum starved and blockage of cholesterol trafficking caused a low rate of phagocytosis and incapability of trophozoites to produce damage in the mouse colon. Ehnpc1 and Ehnpc2 knockdown provoked in trophozoites a lower intracellular cholesterol concentration and a diminished rate of phagocytosis; and Ehnpc1 silencing also produced a decrease of trophozoites movement. Trafficking of EhNPC1 and EhNPC2 during cholesterol uptake and phagocytosis as well as their association with

  7. NPC1 is enriched in unexplained early onset ataxia: a targeted high-throughput screening.

    PubMed

    Synofzik, Matthis; Harmuth, Florian; Stampfer, Miriam; Müller Vom Hagen, Jennifer; Schöls, Ludger; Bauer, Peter

    2015-11-01

    Niemann-Pick disease type C (NP-C) is a rare autosomal-recessive neurodegenerative disease featuring pleiotropic neurological, psychiatric and visceral manifestations. Since many of the adult manifestations can be non-specific or missed, NP-C often goes undetected in adult-onset patients. Here we hypothesized that targeted high-throughput sequencing allows identifying NP-C patients among subjects with unexplained early-onset ataxia (EOA) and, moreover, that this population is enriched for NPC1 mutations. From 204 consecutive EOA patients, all 108 subjects with an established diagnosis were removed (including 4 NPC1 patients), yielding a target cohort of 96 subjects with unexplained EOA, but without primary suspicion of NP-C. This cohort was investigated for NPC1/NPC2 mutations using a high-coverage HaloPlex gene panel including 122 ataxia genes. Among 96 samples, we identified 4 known NPC1 mutations, 3 novel NPC1 missense variants of uncertain significance (VUS) and 1 novel NPC2 missense VUS. The total mutant allele frequency (8/192 = 4.17 %) was significantly enriched compared with control population data (1.57 %; p = 0.011). Two NPC1-positive patients were identified (both with non-specific incipient clinical features), giving a NPC1 patient frequency of 2/96 = 2.1 % in unexplained EOA and of 6/204 = 2.9 % in the total EOA series. NPC1 mutations are substantially enriched in unexplained EOA, demonstrating EOA as a risk-group for NP-C disease. Targeted high-throughput sequencing allows to identify also those NP-C patients with non-specific conditions where the diagnosis has initially been missed. This method does not require having considered NP-C during differential diagnosis, but allows identification of NP-C as part of the default analysis.

  8. Pharmacological specificity of the discriminative stimulus properties of 2-amino-4,5-(1,2-cyclohexyl)-7-phosphono-heptanoic acid (NPC 12626), a competitive N-methyl-D-aspartate receptor antagonist.

    PubMed

    Bobelis, D J; Balster, R L

    1993-02-01

    A drug discrimination based upon the competitive N-methyl-D-aspartate (NMDA) antagonist 2-amino-4,5-(1,2-cyclohexyl)-7-phosphonoheptanoic acid (NPC 12626) was assessed for pharmacological specificity. Adult male Sprague-Dawley rats were trained to discriminate 20 mg/kg i.p. of NPC 12626 from saline under a standard two-lever fixed ratio 32 schedule of food reinforcement. Stimulus generalization tests were conducted to examine the similarities and differences between NPC 12626, its active (2R,4R,5S) enantiomer NPC 17742, other competitive and noncompetitive NMDA antagonists and a number of drugs representative of other classes. During test sessions, the competitive NMDA antagonists NPC 12626, CGS 19755, [1-(cis-2-carboxypiperidine-4-yl)- methyl-1-phosphonic acid], NPC 17742, CSP 37849 [DL-(E)-2-amino-4-methyl-5-phosphono-3-pen-tenoic acid] and CPPene [D-3-(2-carboxypiperazin-4-yl)-1-propenyl-1-phosphonic acid] all completely substituted for the training dose of NPC 12626 with ED50 values of 18.1, 2.3, 2.1, 0.8 and 0.8 mg/kg, respectively. In contrast, drugs that failed to substitute for NPC 12626 included (+)-amphetamine, baclofen, chlorpromazine, dextromethorphan, diazepam, dizocilpine (MK-801), imipramine, (-)-ketocyclazocine, L-N6-phenylisopropyladenosine, methocarbamol, morphine, muscimol, phenytoin, physostigmine and valproate. These results provide evidence that the NPC 12626 discriminative stimulus is unique and specific, shared fully only by its active enantiomer NPC 17742 and other competitive NMDA antagonists. This specificity provides further support for the hypothesis of NMDA receptor mediation of NPC 12626 discrimination, and suggests that this is a useful model to evaluate behavioral effects of competitive NMDA antagonists.

  9. Fomiroid A, a Novel Compound from the Mushroom Fomitopsis nigra, Inhibits NPC1L1-Mediated Cholesterol Uptake via a Mode of Action Distinct from That of Ezetimibe

    PubMed Central

    Chiba, Tomohiro; Sakurada, Tsuyoshi; Watanabe, Rie; Yamaguchi, Kohji; Kimura, Yasuhisa; Kioka, Noriyuki; Kawagishi, Hirokazu; Matsuo, Michinori; Ueda, Kazumitsu

    2014-01-01

    Hypercholesterolemia is one of the key risk factors for coronary heart disease, a major cause of death in developed countries. Suppression of NPC1L1-mediated dietary and biliary cholesterol absorption is predicted to be one of the most effective ways to reduce the risk of hypercholesterolemia. In a screen for natural products that inhibit ezetimibe glucuronide binding to NPC1L1, we found a novel compound, fomiroid A, in extracts of the mushroom Fomitopsis nigra. Fomiroid A is a lanosterone derivative with molecular formula C30H48O3. Fomiroid A inhibited ezetimibe glucuronide binding to NPC1L1, and dose-dependently prevented NPC1L1-mediated cholesterol uptake and formation of esterified cholesterol in NPC1L1-expressing Caco2 cells. Fomiroid A exhibited a pharmacological chaperone activity that corrected trafficking defects of the L1072T/L1168I mutant of NPC1L1. Because ezetimibe does not have such an activity, the binding site and mode of action of fomiroid A are likely to be distinct from those of ezetimibe. PMID:25551765

  10. Quantitative Proteomics of Human Fibroblasts with I1061T Mutation in Niemann–Pick C1 (NPC1) Protein Provides Insights into the Disease Pathogenesis*

    PubMed Central

    Rauniyar, Navin; Subramanian, Kanagaraj; Lavallée-Adam, Mathieu; Martínez-Bartolomé, Salvador; Balch, William E.; Yates, John R.

    2015-01-01

    Niemann-Pick type C (NPC) disease is a fatal neurodegenerative disorder characterized by the accumulation of unesterified cholesterol in the late endosomal/lysosomal compartments. Mutations in the NPC1 protein are implicated in 95% of patients with NPC disease. The most prevalent mutation is the missense mutation I1061T that occurs in ∼15–20% of the disease alleles. In our study, an isobaric labeling-based quantitative analysis of proteome of NPC1I1061T primary fibroblasts when compared with wild-type cells identified 281 differentially expressed proteins based on stringent data analysis criteria. Gene ontology enrichment analysis revealed that these proteins play important roles in diverse cellular processes such as protein maturation, energy metabolism, metabolism of reactive oxygen species, antioxidant activity, steroid metabolism, lipid localization, and apoptosis. The relative expression level of a subset of differentially expressed proteins (TOR4A, DHCR24, CLGN, SOD2, CHORDC1, HSPB7, and GAA) was independently and successfully substantiated by Western blotting. We observed that treating NPC1I1061T cells with four classes of seven different compounds that are potential NPC drugs increased the expression level of SOD2 and DHCR24. We have also shown an abnormal accumulation of glycogen in NPC1I1061T fibroblasts possibly triggered by defective processing of lysosomal alpha-glucosidase. Our study provides a starting point for future more focused investigations to better understand the mechanisms by which the reported dysregulated proteins triggers the pathological cascade in NPC, and furthermore, their effect upon therapeutic interventions. PMID:25873482

  11. Systemic administration of 2-hydroxypropyl-β-cyclodextrin to symptomatic Npc1-deficient mice slows cholesterol sequestration in the major organs and improves liver function.

    PubMed

    Lopez, Adam M; Terpack, Sandi J; Posey, Kenneth S; Liu, Benny; Ramirez, Charina M; Turley, Stephen D

    2014-10-01

    In Niemann-Pick type C (NPC) disease, loss-of-function mutations in either NPC1 or NPC2 result in progressive accumulation of unesterified cholesterol (UC) and glycosphingolipids in all organs, leading to neurodegeneration, pulmonary dysfunction and sometimes liver failure. There is no cure for this disorder. Studies using primarily NPC mouse models have shown that systemic administration of 2-hydroxypropyl-β-cyclodextrin (2HPβCD), starting in early neonatal life, diminishes UC accumulation in most organs, slows disease progression and extends lifespan. The key question now is whether delaying the start of 2HPβCD treatment until early adulthood, when the amount of entrapped UC throughout the body is markedly elevated, has any of the benefits found when treatment begins at 7 days of age. In the present study, Npc1(-/-) and Npc1(+/+) mice were given saline or 2HPβCD subcutaneously at 49, 56, 63 and 70 days of age, with measurements of organ weights, liver function tests and tissue cholesterol levels performed at 77 days. In Npc1(-/-) mice, treatment with 2HPβCD from 49 days reduced whole-liver cholesterol content at 77 days from 33.0 ± 1.0 to 9.1 ± 0.5 mg/organ. Comparable improvements were seen in other organs, such as the spleen, and in the animal as a whole. There was a transient increase in biliary cholesterol concentration in Npc1(-/-) mice after 2HPβCD. Plasma alanine aminotransferase and aspartate aminotransferase activities in 77-day-old 2HPβCD-treated Npc1(-/-) mice were reduced compared with saline-treated controls. The lifespan of Npc1(-/-) mice given 2HPβCD marginally exceeded that of the saline-treated controls (99 ± 1.1 vs 94 ± 1.4 days, respectively; P < 0.05). Thus, 2HPβCD is effective in mobilizing entrapped cholesterol in late-stage NPC disease leading to improved liver function.

  12. Inactivating Mutations in NPC1L1 and Protection from Coronary Heart Disease

    PubMed Central

    2015-01-01

    Background Ezetimibe lowers plasma levels of low-density lipoprotein (LDL) cholesterol by inhibiting the activity of the Niemann–Pick C1-like 1 (NPC1L1) protein. However, whether such inhibition reduces the risk of coronary heart disease is not known. Human mutations that inactivate a gene encoding a drug target can mimic the action of an inhibitory drug and thus can be used to infer potential effects of that drug. Methods We sequenced the exons of NPC1L1 in 7364 patients with coronary heart disease and in 14,728 controls without such disease who were of European, African, or South Asian ancestry. We identified carriers of inactivating mutations (nonsense, splice-site, or frameshift mutations). In addition, we genotyped a specific inactivating mutation (p.Arg406X) in 22,590 patients with coronary heart disease and in 68,412 controls. We tested the association between the presence of an inactivating mutation and both plasma lipid levels and the risk of coronary heart disease. Results With sequencing, we identified 15 distinct NPC1L1 inactivating mutations; approximately 1 in every 650 persons was a heterozygous carrier for 1 of these mutations. Heterozygous carriers of NPC1L1 inactivating mutations had a mean LDL cholesterol level that was 12 mg per deciliter (0.31 mmol per liter) lower than that in noncarriers (P = 0.04). Carrier status was associated with a relative reduction of 53% in the risk of coronary heart disease (odds ratio for carriers, 0.47; 95% confidence interval, 0.25 to 0.87; P = 0.008). In total, only 11 of 29,954 patients with coronary heart disease had an inactivating mutation (carrier frequency, 0.04%) in contrast to 71 of 83,140 controls (carrier frequency, 0.09%). Conclusions Naturally occurring mutations that disrupt NPC1L1 function were found to be associated with reduced plasma LDL cholesterol levels and a reduced risk of coronary heart disease. (Funded by the National Institutes of Health and others.) PMID:25390462

  13. Preclinical evaluation of PI3K inhibitor BYL719 as a single agent and its synergism in combination with cisplatin or MEK inhibitor in nasopharyngeal carcinoma (NPC)

    PubMed Central

    Wong, Chi Hang; Ma, Brigette Buig Yue; Cheong, Hio Teng; Hui, Connie Wun Chun; Hui, Edwin Pun; Chan, Anthony Tak Cheung

    2015-01-01

    Nasopharyngeal carcinoma (NPC) is endemic to Southeast Asia and over 40% of NPC tissues harbor PIK3CA amplifications. This study aims to study the preclinical activity of a novel PI3K inhibitor, BYL719, in 6 NPC cell lines: C666-1, CNE-2, HK1, HK1-EBV, HONE-1 and HONE-1-LMP1. Over 70% of growth inhibition was attained when NPC cell lines were exposed to increasing concentrations of BYL719, with IC50 values at the low micro-molar range. Two BYL719-sensitive cell lines that harbor PIK3CA mutations, CNE-2 and HONE-1, were selected for further analysis on the effect of BYL719 on cell cycle progression, apoptosis and PI3K signaling. BYL719 significantly reduced the phosphorylation of Akt, and the Akt-mTOR axis downstream effector S6 in these 2 cell lines, but a feedback activation of MAPK was observed at 72 hours post-treatment. BYL719 induced G0/G1 cell cycle arrest and apoptosis in both cell lines. In 3D cell culture models, the growth of NPC spheroids was significantly inhibited in a dose-depending manner. When BYL719 was combined with a MEK inhibitor (AZD6244) in a 3D cell culture system, strong synergism on NPC cell growth was observed with attenuation of MAPK activation. A synergistic inhibitory effect on growth was observed when BYL719 was combined with higher dose levels of cisplatin. These data suggest that BYL719 has preclinical activity in NPC cell lines especially in those which harbor PIK3CA mutation. Combination with a MEK inhibitor maybe a useful strategy that warrants further investigation. PMID:26101713

  14. NPC1L1 is a key regulator of intestinal vitamin K absorption and a modulator of warfarin therapy.

    PubMed

    Takada, Tappei; Yamanashi, Yoshihide; Konishi, Kentaro; Yamamoto, Takehito; Toyoda, Yu; Masuo, Yusuke; Yamamoto, Hideaki; Suzuki, Hiroshi

    2015-02-18

    Vitamin K (VK) is a micronutrient that facilitates blood coagulation. VK antagonists, such as warfarin, are used in the clinic to prevent thromboembolism. Because VK is not synthesized in the body, its intestinal absorption is crucial for maintaining whole-body VK levels. However, the molecular mechanism of this absorption is unclear. We demonstrate that Niemann-Pick C1-like 1 (NPC1L1) protein, a cholesterol transporter, plays a central role in intestinal VK uptake and modulates the anticoagulant effect of warfarin. In vitro studies using NPC1L1-overexpressing intestinal cells and in vivo studies with Npc1l1-knockout mice revealed that intestinal VK absorption is NPC1L1-dependent and inhibited by ezetimibe, an NPC1L1-selective inhibitor clinically used for dyslipidemia. In addition, in vivo pharmacological studies demonstrated that the coadministration of ezetimibe and warfarin caused a reduction in hepatic VK levels and enhanced the pharmacological effect of warfarin. Adverse events caused by the coadministration of ezetimibe and warfarin were rescued by oral VK supplementation, suggesting that the drug-drug interaction effects observed were the consequence of ezetimibe-mediated VK malabsorption. This mechanism was supported by a retrospective evaluation of clinical data showing that, in more than 85% of warfarin-treated patients, the anticoagulant activity was enhanced by cotreatment with ezetimibe. Our findings provide insight into the molecular mechanism of VK absorption. This new drug-drug interaction mechanism between ezetimibe (a cholesterol transport inhibitor) and warfarin (a VK antagonist and anticoagulant) could inform clinical care of patients on these medications, such as by altering the kinetics of essential, fat-soluble vitamins.

  15. Neutronics activities for next generation devices

    SciTech Connect

    Gohar, Y.

    1985-01-01

    Neutronic activities for the next generation devices are the subject of this paper. The main activities include TFCX and FPD blanket/shield studies, neutronic aspects of ETR/INTOR critical issues, and neutronics computational modules for the tokamak system code and tandem mirror reactor system code. Trade-off analyses, optimization studies, design problem investigations and computational models development for reactor parametric studies carried out for these activities are summarized.

  16. Rare Loss-of-Function Variants in NPC1 Predispose to Human Obesity.

    PubMed

    Liu, Ruixin; Zou, Yaoyu; Hong, Jie; Cao, Min; Cui, Bin; Zhang, Huiwen; Chen, Maopei; Shi, Juan; Ning, Tinglu; Zhao, Shaoqian; Liu, Wen; Xiong, Hui; Wei, Cuijie; Qiu, Zhengqing; Gu, Weiqiong; Zhang, Yifei; Li, Wanyu; Miao, Lin; Sun, Yingkai; Yang, Minglan; Wang, Rui; Ma, Qinyun; Xu, Min; Xu, Yu; Wang, Tiange; Chan, Kei-Hang Katie; Zuo, Xianbo; Chen, Haoyan; Qi, Lu; Lai, Shenghan; Duan, Shumin; Song, Baoliang; Bi, Yufang; Liu, Simin; Wang, Weiqing; Ning, Guang; Wang, Jiqiu

    2017-04-01

    Some Shanghai Clinical Center f a role of Niemann-Pick type C1 (NPC1) for obesity traits. However, whether the loss-of-function mutations in NPC1 cause adiposity in humans remains unknown. We recruited 25 probands with rare autosomal-recessive Niemann-Pick type C (NP-C) disease and their parents in assessment of the effect of heterozygous NPC1 mutations on adiposity. We found that male NPC1(+/-) carriers had a significantly higher BMI than matched control subjects or the whole population-based control subjects. Consistently, male NPC1(+/-) mice had increased fat storage while eating a high-fat diet. We further conducted an in-depth assessment of rare variants in the NPC1 gene in young, severely obese subjects and lean control subjects and identified 17 rare nonsynonymous/frameshift variants in NPC1 (minor allele frequency <1%) that were significantly associated with an increased risk of obesity (3.40% vs. 0.73%, respectively, in obese patients and control subjects, P = 0.0008, odds ratio = 4.8, 95% CI 1.7-13.2), indicating that rare NPC1 variants were enriched in young, morbidly obese Chinese subjects. Importantly, participants carrying rare variants with severely damaged cholesterol-transporting ability had more fat accumulation than those with mild/no damage rare variants. In summary, rare loss-of-function NPC1 mutations were identified as being associated with human adiposity with a high penetrance, providing potential therapeutic interventions for obesity in addition to the role of NPC1 in the familial NP-C disease.

  17. Npc1 haploinsufficiency promotes weight gain and metabolic features associated with insulin resistance.

    PubMed

    Jelinek, David; Millward, Veronica; Birdi, Amandip; Trouard, Theodore P; Heidenreich, Randall A; Garver, William S

    2011-01-15

    A recent population-based genome-wide association study has revealed that the Niemann-Pick C1 (NPC1) gene is associated with early-onset and morbid adult obesity. Concurrently, our candidate gene-based mouse growth study performed using the BALB/cJ NPC1 mouse model (Npc1) with decreased Npc1 gene dosage independently supported these results by suggesting an Npc1 gene-diet interaction in relation to early-onset weight gain. To further investigate the Npc1 gene in relation to weight gain and metabolic features associated with insulin resistance, we interbred BALB/cJ Npc1(+/-) mice with wild-type C57BL/6J mice, the latter mouse strain commonly used to study aspects of diet-induced obesity and insulin resistance. This breeding produced a hybrid (BALB/cJ-C57BL/6J) Npc1(+/-) mouse model with increased susceptibility to weight gain and insulin resistance. The results from our study indicated that these Npc1(+/-) mice were susceptible to increased weight gain characterized by increased whole body and abdominal adiposity, adipocyte hypertrophy and hepatic steatosis in the absence of hyperphagia. Moreover, these Npc1(+/-) mice developed abnormal metabolic features characterized by impaired fasting glucose, glucose intolerance, hyperinsulinemia, hyperleptinemia and dyslipidemia marked by an increased concentration of cholesterol and triacylglycerol associated with low-density lipoprotein and high-density lipoprotein. The overall results are consistent with a unique Npc1 gene-diet interaction that promotes both weight gain and metabolic features associated with insulin resistance. Therefore, the NPC1 gene now represents a previously unrecognized gene involved in maintaining energy and metabolic homeostasis that will contribute to our understanding concerning the current global epidemic of obesity and type 2 diabetes mellitus.

  18. NREL Pyrheliometer Comparisons: September 26-October 7, 2016 (NPC-2016)

    SciTech Connect

    Reda, Ibrahim; Dooraghi, Mike; Andreas, Afshin; Habte, Aron

    2016-10-01

    Accurate measurements of direct normal (beam) solar irradiance from pyrheliometers are important for developing and deploying solar energy conversion systems, improving our understanding of the Earth's energy budget for climate change studies, and for other science and technology applications involving solar flux. Providing these measurements places many demands on the quality system used by the operator of commercially available radiometers. Maintaining accurate radiometer calibrations that are traceable to an international standard is the first step in producing research-quality solar irradiance measurements. National Renewable Energy Laboratory (NREL) pyrheliometer comparisons (NPCs) are held annually at the Solar Radiation Research Laboratory (SRRL) in Golden, Colorado. Open to all pyrheliometer owners and operators, each NPC provides an opportunity to determine the unique World Radiometric Reference transfer factor (WRR-TF) for each participating pyrheliometer. By adjusting all subsequent pyrheliometer measurements by the appropriate WRR-TF, the solar irradiance data are traceable to the WRR. NPC-2016 was September 26 through October 7, 2016. Participants operated 45 absolute cavity radiometers and 27 conventional thermopile-based pyrheliometers to simultaneously measure clear-sky, direct normal solar irradiance during this period.

  19. NREL Pyrheliometer Comparisons: September 22-26, 2014 (NPC-2014)

    SciTech Connect

    Reda, I.; Dooraghi, M.; Habte, A.

    2014-10-01

    Accurate measurements of direct normal (beam) solar irradiance from pyrheliometers are important for developing and deploying solar energy conversion systems, improving our understanding of the Earth's energy budget for climate change studies, and for other science and technology applications involving solar flux. Providing these measurements places many demands on the quality system used by the operator of commercially available radiometers. Maintaining accurate radiometer calibrations that are traceable to an international standard is the first step in producing research-quality solar irradiance measurements. As with all measurement systems, absolute cavity radiometers and other types of pyrheliometers are subject to performance changes over time. NREL has developed and maintained a select group of absolute cavity radiometers with direct calibration traceability to the World Radiometric Reference (WRR), and uses these reference instruments to calibrate pyrheliometers and pyranometers using the ISO 17025 accredited Broadband Outdoor Radiometer Calibration (BORCAL) process. National Renewable Energy Laboratory (NREL) pyrheliometer comparisons (NPCs) are held annually at the Solar Radiation Research Laboratory (SRRL) in Golden, Colorado. Open to all pyrheliometer owners and operators, each NPC provides an opportunity to determine the unique WRR transfer factor (WRR-TF) for each participating pyrheliometer. By adjusting all subsequent pyrheliometer measurements by the appropriate WRR-TF, the solar irradiance data are traceable to the WRR. NPC-2014 was held September 22-26, 2014. Participants operated 36 absolute cavity radiometers and 22 conventional thermopile-based pyrheliometers to simultaneously measure clear-sky direct normal solar irradiance during this period, and this report documents the findings.

  20. Tetrandrine Induces Apoptosis of Human Nasopharyngeal Carcinoma NPC-TW 076 Cells through Reactive Oxygen Species Accompanied by an Endoplasmic Reticulum Stress Signaling Pathway.

    PubMed

    Lin, Ya-Jing; Peng, Shu-Fen; Lin, Meng-Liang; Kuo, Chao-Lin; Lu, Kung-Wen; Liao, Ching-Lung; Ma, Yi-Shih; Chueh, Fu-Shin; Liu, Kuo-Ching; Yu, Fu-Shun; Chung, Jing-Gung

    2016-10-12

    Nasopharyngeal carcinoma (NPC) is an epithelial malignancy of the head and neck and the incidence is higher in Southeast Asia. Tetrandrine (TET) is a bisbenzylisoquinoline alkaloid, a natural product, and exhibits biological activities including action against many human cancer cell lines. However, the molecular mechanism of TET-induced cell apoptosis in human NPC cells is still unclear. In the present study, we investigated TET-induced apoptotic cell death and associated possible signal pathways on human nasopharyngeal carcinoma NPC-TW 076 cells in vitro. Phase contrast microscopy was used to examine cell morphology and DAPI staining was used to examine chromatin condensation. Flow cytometry assay was used to measure total viable cells, cell cycle and sub-G₁ phase distribution, reactive oxygen species (ROS), Ca(2+), and mitochondria membrane potential (ΔΨm) in NPC-TW 076 cells. Results indicate that TET induced cell death through the cell morphological changes, caused G₀/G₁ phase arrest, increased ROS and Ca(2+) production, and finally caused apoptotic cell death in NPC-TW 076 cells. There was no influence on the level of ΔΨm after TET treatment. Western blotting indicated that TET increased endoplasmic reticulum (ER) stress associated protein expression such as GADD153, GRP78, ATF-6α and ATF-6 βwhich indicated that TET induced cell death through ER stress. ER stress is a potential target in cancer treatment, so the ability of TET to induce ER stress response and to activate programming cell death in NPC-TW 076 cells make this molecule become a promising anticancer agent.

  1. Structure of N-Terminal Domain of NPC1 Reveals Distinct Subdomains for Binding and Transfer of Cholesterol

    SciTech Connect

    Kwon, Hyock Joo; Abi-Mosleh, Lina; Wang, Michael L.; Deisenhofer, Johann; Goldstein, Joseph L.; Brown, Michael S.; Infante, Rodney E.

    2010-09-21

    LDL delivers cholesterol to lysosomes by receptor-mediated endocytosis. Exit of cholesterol from lysosomes requires two proteins, membrane-bound Niemann-Pick C1 (NPC1) and soluble NPC2. NPC2 binds cholesterol with its isooctyl side chain buried and its 3{beta}-hydroxyl exposed. Here, we describe high-resolution structures of the N-terminal domain (NTD) of NPC1 and complexes with cholesterol and 25-hydroxycholesterol. NPC1(NTD) binds cholesterol in an orientation opposite to NPC2: 3{beta}-hydroxyl buried and isooctyl side chain exposed. Cholesterol transfer from NPC2 to NPC1(NTD) requires reorientation of a helical subdomain in NPC1(NTD), enlarging the opening for cholesterol entry. NPC1 with point mutations in this subdomain (distinct from the binding subdomain) cannot accept cholesterol from NPC2 and cannot restore cholesterol exit from lysosomes in NPC1-deficient cells. We propose a working model wherein after lysosomal hydrolysis of LDL-cholesteryl esters, cholesterol binds NPC2, which transfers it to NPC1(NTD), reversing its orientation and allowing insertion of its isooctyl side chain into the outer lysosomal membranes.

  2. Diagnosis of Niemann-Pick disease type C with 7-ketocholesterol screening followed by NPC1/NPC2 gene mutation confirmation in Chinese patients

    PubMed Central

    2014-01-01

    Background It has been reported that oxidation product of cholesterol, 7-ketocholesterol, increases in plasma of patients with NP-C. Previously, we established a rapid test to determine the plasma 7-ketocholesterol level and found it elevated significantly in patients with acid sphingomyelinase deficient NPD and NP-C disease. Methods Individuals randomly referred to our outpatient clinics in the past two years for hepatosplenomegaly or isolated splenomegaly, who have been excluded as acid sphingomyelinase deficient NPD or Gaucher disease, and individuals with newborn cholestasis, psychomotor regression/retardation, were screened for plasma 7-ketocholesterol level. Individuals with high 7-ketocholesterol level were then analyzed for NPC1 and NPC2 gene mutation to confirm the accuracy of NP-C diagnosis. Results By screening the plasma 7-ketocholesterol of suspect individuals, 12 out of 302 (4%) had shown remarkable high levels compared with reference. All these twelve individuals were subsequently confirmed to be NP-C by DNA analysis of NPC1 and NPC2 genes, with the early infantile form (n = 7), the late infantile form (n = 1), the juvenile form (n = 1) and the adult form (n = 1). Furthermore, two NP-C patients without observable neuropsychiatric disability were picked up through this procedure. Only one patient had NP-C due to NPC2 gene mutations, with the rest due to NPC1 gene mutations. We found that in NP-C patients AST was usually mildly elevated and ALT was in a normal range when jaundice was not present. In total, 22 mutant alleles were identified in the NPC1 gene, including six novel small deletions/insertions, e.g., c.416_417insC, c.1030delT, c.1800delC, c.2230_2231delGT, c.2302_2303insG, and c.2795dupA; seven novel exonic point mutations, c.1502A>T (p.D501V), c.1553G>A (p.R518Q), c.1832A>G (p.D611G), c.2054T>C (p.I685T), c.2128C>T(p.Q710X), c.2177G>C (p.R726T), c.2366G>A (p.R789H), and one novel intronic mutation c.2912-3C>G. Small deletions

  3. Identification of NPC1 as the target of U18666A, an inhibitor of lysosomal cholesterol export and Ebola infection.

    PubMed

    Lu, Feiran; Liang, Qiren; Abi-Mosleh, Lina; Das, Akash; De Brabander, Jef K; Goldstein, Joseph L; Brown, Michael S

    2015-12-08

    Niemann-Pick C1 (NPC1) is a lysosomal membrane protein that exports cholesterol derived from receptor-mediated uptake of LDL, and it also mediates cellular entry of Ebola virus. Cholesterol export is inhibited by nanomolar concentrations of U18666A, a cationic sterol. To identify the target of U18666A, we synthesized U-X, a U18666A derivative with a benzophenone that permits ultraviolet-induced crosslinking. When added to CHO cells, U-X crosslinked to NPC1. Crosslinking was blocked by U18666A derivatives that block cholesterol export, but not derivatives lacking blocking activity. Crosslinking was prevented by point mutation in the sterol-sensing domain (SSD) of NPC1, but not by point mutation in the N-terminal domain (NTD). These data suggest that the SSD contains a U18666A-inhibitable site required for cholesterol export distinct from the cholesterol-binding site in the NTD. Inasmuch as inhibition of Ebola requires 100-fold higher concentrations of U18666A, the high affinity U16888A-binding site is likely not required for virus entry.

  4. Generativity as a Route to Active Ageing

    PubMed Central

    Kruse, Andreas; Schmitt, Eric

    2012-01-01

    We elucidate the significance of active ageing from an individual as well as from a societal perspective. Taking an individual perspective, maintaining activity in later years is linked to successful ageing because of empirical relationships to positive self-perception, satisfaction with life, and development of competences, whereas from a societal perspective, active ageing implies usage of older people's life competences as a human capital of society—a societal imperative, particularly in times of demographic change but also more basically substantiated in an ethics of responsibility, intergenerational solidarity, and generation equity. We focus on the psychological construct of generativity which is interpreted as an aspect of the philosophical-anthropological category of joint responsibility. Our own research in Mexico and the Baltic States supports the notion that maintaining access to the public sphere and active engagement for others is a more basic individual concern than a life-stages specific developmental task. We report background and results of a Dialogue Forum Project Funding, a research cooperation between our institute and the Foundation Remembrance, Responsibility, and Future aimed to improve generativity in Belarus, Russia, and Ukraine by implementing and supporting local initiatives offering opportunities for intergenerational dialogue. PMID:22919378

  5. Abnormal gene expression in cerebellum of Npc1-/- mice during postnatal development

    PubMed Central

    Liao, Guanghong; Wen, Zhining; Irizarry, Kristopher; Huang, Ying; Mitsouras, Katherine; Darmani, Mariam; Leon, Terry; Shi, Leming; Bi, Xiaoning

    2010-01-01

    Niemann-Pick Type C disease is an autosomal recessive neurodegenerative disorder with abnormal lipid storage as the major cellular pathologic hallmark. Genetic analyses have identified mutations in NPC1 gene in the great majority of cases, while mutations in NPC2 account for the remainders. Yet, little is known regarding the cellular mechanisms responsible for NPC pathogenesis, especially for neurodegeneration, which is the usual cause of death. To identify critical steps that could account for the pathological manifestations of the disease in one of the most affected brain structures, we performed global gene expression analysis in the cerebellum from three-week old Npc1+/+ and Npc1-/- mice with two different microarray platforms (Agilent and Illumina). Differentially-expressed genes identified by both microarray platforms were then subjected to KEGG pathway analysis. Expression of genes in six pathways was significantly altered in Npc1-/- mice; functionally, these signaling pathways belong to the following three categories: 1) steroid and terpenoid biosynthesis, 2) immune response, and 3) cell adhesion/motility. In addition, the expression of several proteins involved in lipid transport was significantly altered in Npc1-/- mice. Our results provide novel molecular insight regarding the mechanisms of pathogenesis in NPC disease and reveal potential new therapeutic targets. PMID:20153740

  6. Npc1 is involved in sterol trafficking in the filamentous fungus Fusarium graminearum

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The ortholog of the human gene NPC1 was identified in the plant pathogenic, filamentous fungus Fusarium graminearum by shared amino acid sequence, protein domain structure and cellular localization of the mature fungal protein. The Fusarium Npc1 gene shares 34% amino acid sequence identity and 51% s...

  7. Swim pressure: stress generation in active matter.

    PubMed

    Takatori, S C; Yan, W; Brady, J F

    2014-07-11

    We discover a new contribution to the pressure (or stress) exerted by a suspension of self-propelled bodies. Through their self-motion, all active matter systems generate a unique swim pressure that is entirely athermal in origin. The origin of the swim pressure is based upon the notion that an active body would swim away in space unless confined by boundaries-this confinement pressure is precisely the swim pressure. Here we give the micromechanical basis for the swim stress and use this new perspective to study self-assembly and phase separation in active soft matter. The swim pressure gives rise to a nonequilibrium equation of state for active matter with pressure-volume phase diagrams that resemble a van der Waals loop from equilibrium gas-liquid coexistence. Theoretical predictions are corroborated by Brownian dynamics simulations. Our new swim stress perspective can help analyze and exploit a wide class of active soft matter, from swimming bacteria to catalytic nanobots to molecular motors that activate the cellular cytoskeleton.

  8. Active Ageing: Intergenerational Relationships and Social Generativity.

    PubMed

    Rossi, Giovanna; Boccacin, Lucia; Bramanti, Donatella; Meda, Stefania G

    2014-01-01

    This contribution is a reflection on the concept of active ageing from the perspective of relational sociology. At the same time, it offers practical implications and outlines possible future courses of action, in the face of demographic and relational scenarios rapidly changing, and the challenges that each day people of all generations are called to cope with. Active ageing is quite a recent concept and indicates an attitude towards ageing that enhances the quality of life as people become older. The goal of active ageing is to enable people to realise their potential for physical, social and mental well-being and to participate in social life also in the last stage of the life cycle. In this phase, the presence of a network of support, security and care adequate to the possible onset of problems and criticalities is crucial. Relational sociology frames the phenomenon of an ageing population in a dense network of social relations, primarily at the level of family and community. For this reason, as supported by the most recent sociological literature and evidence from studies conducted in Italy and abroad (cf. SHARE), it is extremely important to investigate the link between active ageing, intergenerational orientation (solidarity and exchanges) and practices of prosociality (i.e. engagement in third-sector activities and volunteering in later life).

  9. Cytoplasmic Drosha activity generated by alternative splicing

    PubMed Central

    Dai, Lisheng; Chen, Kevin; Youngren, Brenda; Kulina, Julia; Yang, Acong; Guo, Zhengyu; Li, Jin; Yu, Peng; Gu, Shuo

    2016-01-01

    RNase III enzyme Drosha interacts with DGCR8 to form the Microprocessor, initiating canonical microRNA (miRNA) maturation in the nucleus. Here, we re-evaluated where Drosha functions in cells using Drosha and/or DGCR8 knock out (KO) cells and cleavage reporters. Interestingly, a truncated Drosha mutant located exclusively in the cytoplasm cleaved pri-miRNA effectively in a DGCR8-dependent manner. In addition, we demonstrated that in vitro generated pri-miRNAs when transfected into cells could be processed to mature miRNAs in the cytoplasm. These results indicate the existence of cytoplasmic Drosha (c-Drosha) activity. Although a subset of endogenous pri-miRNAs become enriched in the cytoplasm of Drosha KO cells, it remains unclear whether pri-miRNA processing is the main function of c-Drosha. We identified two novel in-frame Drosha isoforms generated by alternative splicing in both HEK293T and HeLa cells. One isoform loses the putative nuclear localization signal, generating c-Drosha. Further analysis indicated that the c-Drosha isoform is abundant in multiple cell lines, dramatically variable among different human tissues and upregulated in multiple tumors, suggesting that c-Drosha plays a unique role in gene regulation. Our results reveal a new layer of regulation on the miRNA pathway and provide novel insights into the ever-evolving functions of Drosha. PMID:27471035

  10. NREL Pyrheliometer Comparison: September 16 to 27, 2013 (NPC-2013)

    SciTech Connect

    Reda, I.; Dooraghi, M.; Habte, A.

    2013-11-01

    Accurate measurements of direct normal (beam) solar irradiance from pyrheliometers are important for the development and deployment of solar energy conversion systems, improving our understanding of the Earth's energy budget for climate change studies, and for other science and technology applications involving solar flux. Providing these measurements places many demands on the quality system used by the operator of commercially available radiometers. Maintaining accurate radiometer calibrations traceable to an international standard is the first step in producing research-quality solar irradiance measurements. As with all measurement systems, absolute cavity radiometers and other types of pyrheliometers are subject to performance changes over time. NREL has developed and maintained a group of absolute cavity radiometers with direct calibration traceability to the World Radiometric Reference (WRR). These reference instruments are used by NREL to calibrate pyrheliometers and pyranometers using the ISO 17025 accredited Broadband Outdoor Radiometer Calibration (BORCAL) process (Reda et al. 2008). NPCs are held annually at the Solar Radiation Research Laboratory (SRRL) in Golden, Colorado. Open to all pyrheliometer owners/operators, e.g. NREL, NASA, NIST, NOAA, USA industry and academia, USA-DOE and other national laboratories, and national and international organizations. Each NPC provides an opportunity to determine the unique World Radiometric Reference (WRR) transfer factor (WRR-TF) for each participating pyrheliometer. By adjusting all subsequent pyrheliometer measurements by the appropriate WRR-TF, the solar irradiance data are traceable to the International System of Units through WRR.

  11. A Single Residue in Ebola Virus Receptor NPC1 Influences Cellular Host Range in Reptiles

    DTIC Science & Technology

    2016-09-07

    particles bearing cleaved EBOV GP (rVSV-GPCL) as 116   measured by ELISA(22); however, no ELISA signal was apparent even at the highest concentra-117... bearing amino acid residues with divergent physicochemical properties at position 503. Exami-224   nation of these mutants by GPCL binding ELISA...bidirectionally alters domain C’s capacity to bind EBOV GPCL. (A) HsNPC1 and DrNPC1 domain C proteins bearing point mutations at residue 503 (HsNPC1, F503Y

  12. 20 CFR 627.225 - Employment generating activities.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... generating activities, economic development activities, investment in revolving loan funds, capitalization of... activities, economic development activities, or similar activities. (b) JTPA funds may be used for normal... chambers of commerce); JTPA staff participation on economic development boards and commissions, and...

  13. Colony Rheology: Active Arthropods Generate Flows

    NASA Astrophysics Data System (ADS)

    Daniels, Karen; Mann, Michael; Charbonneau, Patrick

    2015-03-01

    Hydrodynamic-like flows are observed in biological systems as varied as bacteria, insects, birds, fish, and mammals. Both the phenomenology (e.g. front instabilities, milling motions) and the interaction types (hydrodynamic, direct contact, psychological, excluded-volume) strongly vary between systems, but a question common to all of them is to understand the role of particle-scale fluctuations in controlling large-scale rheological behaviors. We will address these questions through experiments on a new system, Tyrolichus casei (cheese mites), which live in dense, self-mixing colonies composed of a mixture of living mites and inert flour/detritus. In experiments performed in a Hele-Shaw geometry, we observe that the rheology of a colony is strongly dependent on the relative concentration of active and inactive particles. In addition to spreading flows, we also observe that the system can generate convective circulation and auto-compaction.

  14. T cell activation requires force generation

    PubMed Central

    Hu, Kenneth H.

    2016-01-01

    Triggering of the T cell receptor (TCR) integrates both binding kinetics and mechanical forces. To understand the contribution of the T cell cytoskeleton to these forces, we triggered T cells using a novel application of atomic force microscopy (AFM). We presented antigenic stimulation using the AFM cantilever while simultaneously imaging with optical microscopy and measuring forces on the cantilever. T cells respond forcefully to antigen after calcium flux. All forces and calcium responses were abrogated upon treatment with an F-actin inhibitor. When we emulated the forces of the T cell using the AFM cantilever, even these actin-inhibited T cells became activated. Purely mechanical stimulation was not sufficient; the exogenous forces had to couple through the TCR. These studies suggest a mechanical–chemical feedback loop in which TCR-triggered T cells generate forceful contacts with antigen-presenting cells to improve access to antigen. PMID:27241914

  15. NREL Pyrheliometer Comparisons (NPC-2001), September 24 - October 5, 2001, Golden, Colorado

    SciTech Connect

    Reda, I.; Stoffel, T.; Wilcox, S.

    2003-09-01

    NREL Pyrheliometer Comparisons (NPCs) are held annually at the Solar Radiation Research Laboratory (SRRL) in Golden, Colorado. Open to all pyrheliometer owner/operators, the NPC provides an opportunity to determine the unique WRR transfer factor for each participating pyrheliometer.

  16. Results of NREL Pyrheliometer Comparisons (NPC1999), October 4-10, 1999, Golden, Colorado

    SciTech Connect

    Reda, I.; Stoffel, T.; Wilcox, S.

    2000-09-01

    NREL Pyrheliometer Comparisons (NPCs) are held annually at the Solar Radiation Research Laboratory (SRRL) in Golden, Colorado. Open to all pyrheliometer owner/operators, the NPC provides an opportunity to determine the unique WRR transfer factor for each participating pyrheliometer.

  17. NREL Pyrheliometer Comparisons (NPC-2002), September 23 - October 4, 2002, Golden, Colorado

    SciTech Connect

    Reda, I.; Stoffel, T.; Wilcox, S.

    2003-09-01

    NREL Pyrheliometer Comparisons (NPCs) are held annually at the Solar Radiation Research Laboratory (SRRL) in Golden, Colorado. Open to all pyrheliometer owner/operators, the NPC provides an opportunity to determine the unique WRR transfer factor for each participating pyrheliometer.

  18. NREL Pyrheliometer Comparisons (NPC-2003), September 22 - October 3, 2003, Golden, Colorado

    SciTech Connect

    Stoffel, T.; Reda, I.; Wilcox, S.

    2004-07-01

    NREL Pyrheliometer Comparisons (NPCs) are held annually at the Solar Radiation Research Laboratory (SRRL) in Golden, Colorado. Open to all pyrheliometer owner/operators, the NPC provides an opportunity to determine the unique WRR transfer factor for each participating pyrheliometer.

  19. 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) induced P-450 mediated arachidonic acid (AA) metabolism in chick embryo liver (CEL) occurs in parenchymal cells (PC) rather than in non-parenchymal cells (NPC)

    SciTech Connect

    Paroli, L.; Rifkind, A.B. )

    1992-02-26

    TCDD induces cytochrome P-450 mediated AA metabolism in CEL and changes the dominant metabolite(s) from {omega}-OH AA to AA epoxygenase products (EETs and EET-diols). PC and NPC from CEL were separated by differential centrifugation and characterized by morphology, immunohistochemistry and P-450 mediated xenobiotic metabolism; purities were >95%. PC and NPC, from 16 day old chick embryos treated for 5 days with TCDD or vehicle alone, were cultured for 48 hr, homogenized and incubated with ({sup 14}C)-AA {plus minus} NADPH. AA products were resolved by reverse phase HPLC. The major product in control PC, {omega}-OH AA was not significantly affected by TCDD. All of the AA metabolism was NADPH dependent. Control and TCDD treated PC had the same metabolite patterns as whole liver microsomes. Neither control nor TCDD treated NPC generated P-450 AA metabolites. Also co-culturing NPC with PC did not affect AA metabolism of either cell type. The findings indicate that TCDD-induced changes in AA metabolism are retained in culture and that hepatocytes rather than NPC effect P-450 mediated AA metabolism in both control and TCDD-induced CEL.

  20. [Statistical tests in medical research: traditional methods vs. multivariate NPC permutation tests].

    PubMed

    Arboretti, Rosa; Bordignon, Paolo; Corain, Livio; Palermo, Giuseppe; Pesarin, Fortunato; Salmaso, Luigi

    2015-01-01

    Statistical tests in medical research: traditional methods vs. multivariate npc permutation tests.Within medical research, a useful statistical tool is based on hypotheses testing in terms of the so-called null, that is the treatment has no effect, and alternative hypotheses, that is the treatment has some effects. By controlling the risks of wrong decisions, empirical data are used in order to possibly reject the null hypotheses in favour of the alternative, so that demonstrating the efficacy of a treatment of interest. The multivariate permutation tests, based on the nonparametric combination - NPC method, provide an innovative, robust and effective hypotheses testing solution to many real problems that are commonly encountered in medical research when multiple end-points are observed. This paper discusses the various approaches to hypothesis testing and the main advantages of NPC tests, which consist in the fact that they require much less stringent assumptions than traditional statistical tests. Moreover, the related results may be extended to the reference population even in case of selection-bias, that is non-random sampling. In this work, we review and discuss some basic testing procedures along with the theoretical and practical relevance of NPC tests showing their effectiveness in medical research. Within the non-parametric methods, NPC tests represent the current "frontier" of statistical research, but already widely available in the practice of analysis of clinical data.

  1. Filovirus receptor NPC1 contributes to species-specific patterns of ebolavirus susceptibility in bats

    PubMed Central

    Ng, Melinda; Ndungo, Esther; Kaczmarek, Maria E; Herbert, Andrew S; Binger, Tabea; Kuehne, Ana I; Jangra, Rohit K; Hawkins, John A; Gifford, Robert J; Biswas, Rohan; Demogines, Ann; James, Rebekah M; Yu, Meng; Brummelkamp, Thijn R; Drosten, Christian; Wang, Lin-Fa; Kuhn, Jens H; Müller, Marcel A; Dye, John M; Sawyer, Sara L; Chandran, Kartik

    2015-01-01

    Biological factors that influence the host range and spillover of Ebola virus (EBOV) and other filoviruses remain enigmatic. While filoviruses infect diverse mammalian cell lines, we report that cells from African straw-colored fruit bats (Eidolon helvum) are refractory to EBOV infection. This could be explained by a single amino acid change in the filovirus receptor, NPC1, which greatly reduces the affinity of EBOV-NPC1 interaction. We found signatures of positive selection in bat NPC1 concentrated at the virus-receptor interface, with the strongest signal at the same residue that controls EBOV infection in Eidolon helvum cells. Our work identifies NPC1 as a genetic determinant of filovirus susceptibility in bats, and suggests that some NPC1 variations reflect host adaptations to reduce filovirus replication and virulence. A single viral mutation afforded escape from receptor control, revealing a pathway for compensatory viral evolution and a potential avenue for expansion of filovirus host range in nature. DOI: http://dx.doi.org/10.7554/eLife.11785.001 PMID:26698106

  2. Structural Insights into the Niemann-Pick C1 (NPC1)-Mediated Cholesterol Transfer and Ebola Infection.

    PubMed

    Gong, Xin; Qian, Hongwu; Zhou, Xinhui; Wu, Jianping; Wan, Tao; Cao, Pingping; Huang, Weiyun; Zhao, Xin; Wang, Xudong; Wang, Peiyi; Shi, Yi; Gao, George F; Zhou, Qiang; Yan, Nieng

    2016-06-02

    Niemann-Pick disease type C (NPC) is associated with mutations in NPC1 and NPC2, whose gene products are key players in the endosomal/lysosomal egress of low-density lipoprotein-derived cholesterol. NPC1 is also the intracellular receptor for Ebola virus (EBOV). Here, we present a 4.4 Å structure of full-length human NPC1 and a low-resolution reconstruction of NPC1 in complex with the cleaved glycoprotein (GPcl) of EBOV, both determined by single-particle electron cryomicroscopy. NPC1 contains 13 transmembrane segments (TMs) and three distinct lumenal domains A (also designated NTD), C, and I. TMs 2-13 exhibit a typical resistance-nodulation-cell division fold, among which TMs 3-7 constitute the sterol-sensing domain conserved in several proteins involved in cholesterol metabolism and signaling. A trimeric EBOV-GPcl binds to one NPC1 monomer through the domain C. Our structural and biochemical characterizations provide an important framework for mechanistic understanding of NPC1-mediated intracellular cholesterol trafficking and Ebola virus infection.

  3. Estimating evaporative vapor generation from automobiles based on parking activities.

    PubMed

    Dong, Xinyi; Tschantz, Michael; Fu, Joshua S

    2015-07-01

    A new approach is proposed to quantify the evaporative vapor generation based on real parking activity data. As compared to the existing methods, two improvements are applied in this new approach to reduce the uncertainties: First, evaporative vapor generation from diurnal parking events is usually calculated based on estimated average parking duration for the whole fleet, while in this study, vapor generation rate is calculated based on parking activities distribution. Second, rather than using the daily temperature gradient, this study uses hourly temperature observations to derive the hourly incremental vapor generation rates. The parking distribution and hourly incremental vapor generation rates are then adopted with Wade-Reddy's equation to estimate the weighted average evaporative generation. We find that hourly incremental rates can better describe the temporal variations of vapor generation, and the weighted vapor generation rate is 5-8% less than calculation without considering parking activity.

  4. Triazoles inhibit cholesterol export from lysosomes by binding to NPC1.

    PubMed

    Trinh, Michael N; Lu, Feiran; Li, Xiaochun; Das, Akash; Liang, Qiren; De Brabander, Jef K; Brown, Michael S; Goldstein, Joseph L

    2017-01-03

    Niemann-Pick C1 (NPC1), a membrane protein of lysosomes, is required for the export of cholesterol derived from receptor-mediated endocytosis of LDL. Lysosomal cholesterol export is reportedly inhibited by itraconazole, a triazole that is used as an antifungal drug [Xu et al. (2010) Proc Natl Acad Sci USA 107:4764-4769]. Here we show that posaconazole, another triazole, also blocks cholesterol export from lysosomes. We prepared P-X, a photoactivatable cross-linking derivative of posaconazole. P-X cross-linked to NPC1 when added to intact cells. Cross-linking was inhibited by itraconazole but not by ketoconazole, an imidazole that does not block cholesterol export. Cross-linking of P-X was also blocked by U18666A, a compound that has been shown to bind to NPC1 and inhibit cholesterol export. P-X also cross-linked to purified NPC1 that was incorporated into lipid bilayer nanodiscs. In this in vitro system, cross-linking of P-X was inhibited by itraconazole, but not by U18666A. P-X cross-linking was not prevented by deletion of the N-terminal domain of NPC1, which contains the initial binding site for cholesterol. In contrast, P-X cross-linking was reduced when NPC1 contained a point mutation (P691S) in its putative sterol-sensing domain. We hypothesize that the sterol-sensing domain has a binding site that can accommodate structurally different ligands.

  5. Fluidic Active Transducer for Electricity Generation

    PubMed Central

    Yang, YoungJun; Park, Junwoo; Kwon, Soon-Hyung; Kim, Youn Sang

    2015-01-01

    Flows in small size channels have been studied for a long time over multidisciplinary field such as chemistry, biology and medical through the various topics. Recently, the attempts of electricity generation from the small flows as a new area for energy harvesting in microfluidics have been reported. Here, we propose for the first time a new fluidic electricity generator (FEG) by modulating the electric double layer (EDL) with two phase flows of water and air without external power sources. We find that an electric current flowed by the forming/deforming of the EDL with a simple separated phase flow of water and air at the surface of the FEG. Electric signals between two electrodes of the FEG are checked from various water/air passing conditions. Moreover, we verify the possibility of a self-powered air slug sensor by applying the FEG in the detection of an air slug. PMID:26511626

  6. Fluidic Active Transducer for Electricity Generation.

    PubMed

    Yang, YoungJun; Park, Junwoo; Kwon, Soon-Hyung; Kim, Youn Sang

    2015-10-29

    Flows in small size channels have been studied for a long time over multidisciplinary field such as chemistry, biology and medical through the various topics. Recently, the attempts of electricity generation from the small flows as a new area for energy harvesting in microfluidics have been reported. Here, we propose for the first time a new fluidic electricity generator (FEG) by modulating the electric double layer (EDL) with two phase flows of water and air without external power sources. We find that an electric current flowed by the forming/deforming of the EDL with a simple separated phase flow of water and air at the surface of the FEG. Electric signals between two electrodes of the FEG are checked from various water/air passing conditions. Moreover, we verify the possibility of a self-powered air slug sensor by applying the FEG in the detection of an air slug.

  7. Structural Basis of Sterol Binding by NPC2, a Lysosomal Protein Deficient in Niemann-Pick Type C2 Disease

    SciTech Connect

    Xu,S.; Benoff, B.; Liou, H.; Lobel, P.; Stock, A.

    2007-01-01

    NPC2 is a small lysosomal glycoprotein that binds cholesterol with submicromolar affinity. Deficiency in NPC2 is the cause of Niemann-Pick type C2 disease, a fatal neurovisceral disorder characterized by accumulation of cholesterol in lysosomes. Here we report the crystal structure of bovine NPC2 bound to cholesterol-3-O-sulfate, an analog that binds with greater apparent affinity than cholesterol. Structures of both apo-bound and sterol-bound NPC2 were observed within the same crystal lattice, with an asymmetric unit containing one molecule of apoNPC2 and two molecules of sterol-bound NPC2. As predicted from a previously determined structure of apoNPC2, the sterol binds in a deep hydrophobic pocket sandwiched between the two {beta}-sheets of NPC2, with only the sulfate substituent of the ligand exposed to solvent. In the two available structures of apoNPC2, the incipient ligand-binding pocket, which ranges from a loosely packed hydrophobic core to a small tunnel, is too small to accommodate cholesterol. In the presence of sterol, the pocket expands, facilitated by a slight separation of the {beta}-strands and substantial reorientation of some side chains, resulting in a perfect molding of the pocket around the hydrocarbon portion of cholesterol. A notable feature is the repositioning of two aromatic residues at the tunnel entrance that are essential for NPC2 function. The NPC2 structures provide evidence of a malleable binding site, consistent with the previously documented broad range of sterol ligand specificity.

  8. STRUCTURAL BASIS OF STEROL BINDING BY NPC2, A LYSOSOMAL PROTEIN DEFICIENT IN NIEMANN-PICK TYPE C2 DISEASE*

    PubMed Central

    Xu, Sujuan; Benoff, Brian; Liou, Heng-Ling; Lobel, Peter; Stock, Ann M.

    2013-01-01

    NPC2 is a small lysosomal glycoprotein that binds cholesterol with submicromolar affinity. Deficiency in NPC2 is the cause of Niemann Pick type C2 disease, a fatal neurovisceral disorder characterized by accumulation of cholesterol in lysosomes. Here we report the crystal structure of bovine NPC2 bound to cholesterol-3-O-sulfate, an analog that binds with greater apparent affinity than cholesterol. Structures of both apo- and sterol-bound NPC2 were observed within the same crystal lattice, with an asymmetric unit containing one molecule of apoNPC2 and two molecules of sterol-bound NPC2. As predicted from a previously determined structure of apoNPC2, the sterol binds in a deep hydrophobic pocket sandwiched between the two β sheets of NPC2, with only the sulfate substituent of the ligand exposed to solvent. In the two available structures of apoNPC2, the incipient ligand-binding pocket, which ranges from a loosely packed hydrophobic core to a small tunnel, is too small to accommodate cholesterol. In the presence of sterol, the pocket expands, facilitated by a slight separation of the β strands and substantial reorientation of some side chains, resulting in a perfect molding of the pocket around the hydrocarbon portion of cholesterol. A notable feature is the repositioning of two aromatic residues at the tunnel entrance that are essential for NPC2 function. The NPC2 structures provide evidence of a malleable binding site, consistent with the previously documented broad range of sterol ligand specificity. PMID:17573352

  9. Cholesterol accumulation in Niemann Pick type C (NPC) model cells causes a shift in APP localization to lipid rafts

    SciTech Connect

    Kosicek, Marko; Malnar, Martina; Goate, Alison; Hecimovic, Silva

    2010-03-12

    It has been suggested that cholesterol may modulate amyloid-{beta} (A{beta}) formation, a causative factor of Alzheimer's disease (AD), by regulating distribution of the three key proteins in the pathogenesis of AD ({beta}-amyloid precursor protein (APP), {beta}-secretase (BACE1) and/or presenilin 1 (PS1)) within lipid rafts. In this work we tested whether cholesterol accumulation upon NPC1 dysfunction, which causes Niemann Pick type C disease (NPC), causes increased partitioning of APP into lipid rafts leading to increased CTF/A{beta} formation in these cholesterol-rich membrane microdomains. To test this we used CHO NPC1{sup -/-} cells (NPC cells) and parental CHOwt cells. By sucrose density gradient centrifugation we observed a shift in fl-APP/CTF compartmentalization into lipid raft fractions upon cholesterol accumulation in NPC vs. wt cells. Furthermore, {gamma}-secretase inhibitor treatment significantly increased fl-APP/CTF distribution in raft fractions in NPC vs. wt cells, suggesting that upon cholesterol accumulation in NPC1-null cells increased formation of APP-CTF and its increased processing towards A{beta} occurs in lipid rafts. Our results support that cholesterol overload, such as in NPC disease, leads to increased partitioning of APP/CTF into lipid rafts resulting in increased amyloidogenic processing of APP in these cholesterol-rich membranes. This work adds to the mechanism of the cholesterol-effect on APP processing and the pathogenesis of Alzheimer's disease and supports the role of lipid rafts in these processes.

  10. Next Generation Active Buffet Suppression System

    NASA Technical Reports Server (NTRS)

    Galea, Stephen C.; Ryall, Thomas G.; Henderson, Douglas A.; Moses, Robert W.; White, Edward V.; Zimcik, David G.

    2003-01-01

    Buffeting is an aeroelastic phenomenon that is common to high performance aircraft, especially those with twin vertical tails like the F/A-18, at high angles of attack. These loads result in significant random stresses, which may cause fatigue damage leading to restricted capabilities and availability of the aircraft. This paper describes an international collaborative research activity among Australia, Canada and the United States involving the use of active structural control to alleviate the damaging structural response to these loads. The research program is being co-ordinated by the Air Force Research Laboratory (AFRL) and is being conducted under the auspices of The Technical Cooperative Program (TTCP). This truly unique collaborative program has been developed to enable each participating country to contribute resources toward a program that coalesces a broad range of technical knowledge and expertise into a single investigation. This collaborative program is directed toward a full-scale test of an F/A-18 empennage, which is an extension of an earlier initial test. The current program aims at applying advanced directional piezoactuators, the aircraft rudder, switch mode amplifiers and advanced control strategies on a full-scale structure to demonstrate the enhanced performance and capability of the advanced active BLA control system in preparation for a flight test demonstration.

  11. National Renewable Energy Laboratory Pyrheliometer Comparisons: 24 September - 5 October 2012 (NPC-2012)

    SciTech Connect

    Stoffel, T.; Reda, I.

    2013-05-01

    The NREL Pyrheliometer Comparisons for 2012 (NPC-2012) were held at the Solar Radiation Research Laboratory in Golden, Colorado, from September 24 through October 5 for the purpose of transferring the World Radiometric Reference (WRR) to participating instrument. Twenty scientists and engineers operated 32 absolute cavity radiometers and 18 conventional thermopile-based pyrheliometers to simultaneously measure clear-sky direct normal irradiance during the comparisons. The transfer standard group of reference radiometers for NPC-2012 consisted of four NREL radiometers with direct traceability to the WRR, having participated in the Eleventh International Pyrheliometer Comparisons (IPC-XI) hosted by the World Radiation Center in the fall of 2010. As the result of NPC-2012, each participating absolute cavity radiometer was assigned a new WRR transfer factor, computed as the reference irradiance computed by the transfer standard group divided by the observed irradiance from the participating radiometer. The performance of the transfer standard group during NPC-2012 was consistent with previous comparisons, including IPC-XI. The measurement performance of the transfer standard group allowed the transfer of the WRR to each participating radiometer with an estimated uncertainty of +/- 0.33% with respect to the International System of Units.

  12. Amelioration of non-alcoholic fatty liver disease with NPC1L1-targeted IgY or n-3 polyunsaturated fatty acids in mice.

    PubMed

    Bae, Jin-Sik; Park, Jong-Min; Lee, Junghoon; Oh, Byung-Chul; Jang, Sang-Ho; Lee, Yun Bin; Han, Young-Min; Ock, Chan-Young; Cha, Ji-Young; Hahm, Ki-Baik

    2017-01-01

    Patients with non-alcoholic fatty liver disease (NAFLD) have an increased risk for progression to hepatocellular carcinoma in addition to comorbidities such as cardiovascular and serious metabolic diseases; however, the current therapeutic options are limited. Based on our previous report that omega-3 polyunsaturated fatty acids (n-3 PUFAs) can significantly ameliorate high fat diet (HFD)-induced NAFLD, we explored the therapeutic efficacy of n-3 PUFAs and N-IgY, which is a chicken egg yolk-derived IgY specific for the Niemann-Pick C1-Like 1 (NPC1L1) cholesterol transporter, on NAFLD in mice. We generated N-IgY and confirmed its efficient cholesterol transport-blocking activity in HepG2 and Caco-2 cells, which was comparable to the effect of ezetimibe (EZM). C57BL/6 wild type and fat-1 transgenic mice, capable of producing n-3 PUFAs, were fed a high fat diet (HFD) alone or supplemented with N-IgY. Endogenously synthesized n-3 PUFAs combined with N-IgY led to significant decreases in hepatic steatosis, fibrosis, and inflammation (p<0.01). The combination of N-IgY and n-3 PUFAs resulted in significant upregulation of genes involved in cholesterol uptake (LDLR), reverse cholesterol transport (ABCG5/ABCG8), and bile acid metabolism (CYP7A1). Moreover, fat-1 transgenic mice treated with N-IgY showed significant downregulation of genes involved in cholesterol-induced hepatic stellate cell activation (Tgfb1, Tlr4, Col1a1, Col1a2, and Timp2). Collectively, these data suggest that n-3 PUFAs and N-IgY, alone or in combination, represent a promising treatment strategy to prevent HFD-induced fatty liver through the activation cholesterol catabolism to bile acids and by decreasing cholesterol-induced fibrosis.

  13. Loss of Cathepsin B and L Leads to Lysosomal Dysfunction, NPC-Like Cholesterol Sequestration and Accumulation of the Key Alzheimer's Proteins

    PubMed Central

    Cermak, Stjepko; Kosicek, Marko; Mladenovic-Djordjevic, Aleksandra; Smiljanic, Kosara; Kanazir, Selma

    2016-01-01

    Proper function of lysosomes is particularly important in neurons, as they cannot dilute accumulated toxic molecules and aggregates by cell division. Thus, impairment of lysosomal function plays an important role in neuronal degeneration and in the pathogenesis of numerous neurodegenerative diseases. In this work we analyzed how inhibition and/or loss of the major lysosomal proteases, the cysteine cathepsins B and L (CtsB/L), affects lysosomal function, cholesterol metabolism and degradation of the key Alzheimer's disease (AD) proteins. Here, we show that cysteine CtsB/L, and not the aspartyl cathepsin D (CtsD), represent a major lysosomal protease(s) that control lysosomal function, intracellular cholesterol trafficking and AD-like amyloidogenic features. Intriguingly, accumulation of free cholesterol in late endosomes/lysosomes upon CtsB/L inhibition resembled a phenotype characteristic for the rare neurodegenerative disorder Niemann-Pick type C (NPC). CtsB/L inhibition and not the inhibition of CtsD led to lysosomal impairment assessed by decreased degradation of EGF receptor, enhanced LysoTracker staining and accumulation of several lysosomal proteins LC3II, NPC1 and NPC2. By measuring the levels of NPC1 and ABCA1, the two major cholesterol efflux proteins, we showed that CtsB/L inhibition or genetic depletion caused accumulation of the NPC1 in lysosomes and downregulation of ABCA1 protein levels and its expression. Furthermore, we revealed that CtsB/L are involved in degradation of the key Alzheimer’s proteins: amyloid-β peptides (Aβ) and C-terminal fragments of the amyloid precursor protein (APP) and in degradation of β-secretase (BACE1). Our results imply CtsB/L as major regulators of lysosomal function and demonstrate that CtsB/L may play an important role in intracellular cholesterol trafficking and in degradation of the key AD proteins. Our findings implicate that enhancing the activity or levels of CtsB/L could provide a promising and a common

  14. NASA/NREN: Next Generation Internet (NGI) Activities

    NASA Technical Reports Server (NTRS)

    desJardins, Richard; Freeman, Ken

    1998-01-01

    Various issues associated with next generation internet (NGI) and the NREN (NASA Research and Education Network) activities are presented in viewgraph form. Specific topics include: 1) NREN architecture; 2) NREN applications; and 3) NREN applied research.

  15. Application of Co-naphthalocyanine (CoNPc) as alternative cathode catalyst and support structure for microbial fuel cells.

    PubMed

    Kim, Jung Rae; Kim, Jy-Yeon; Han, Sang-Beom; Park, Kyung-Won; Saratale, G D; Oh, Sang-Eun

    2011-01-01

    Co-naphthalocyanine (CoNPc) was prepared by heat treatment for cathode catalysts to be used in microbial fuel cells (MFCs). Four different catalysts (Carbon black, NPc/C, CoNPc/C, Pt/C) were compared and characterized using XPS, EDAX and TEM. The electrochemical characteristics of oxygen reduction reaction (ORR) were compared by cyclic voltammetry (CV) and linear sweep voltammetry (LSV). The Co-macrocyclic complex improves the catalyst dispersion and oxygen reduction reaction of CoNPc/C. The maximum power of CoNPc/C was 64.7 mW/m(2) at 0.25 mA as compared with 81.3 mW/m(2) of Pt/C, 29.7 mW/m(2) of NPc/C and 9.3 mW/m(2) of carbon black when the cathodes were implemented in H-type MFCs. The steady state cell, cathode and anode potential of MFC with using CoNPc/C were comparable to those of Pt/C.

  16. Pulsed neutron generator for use with pulsed neutron activation techniques

    SciTech Connect

    Rochau, G.E.

    1980-01-01

    A high-output, transportable, pulsed neutron generator has been developed by Sandia National Laboratories for use with Pulsed Neutron Activation (PNA) techniques. The PNA neutron generator generates > 10/sup 10/ 14 MeV D-T neutrons in a 1.2 millisecond pulse. Each operation of the unit will produce a nominal total neutron output of 1.2 x 10/sup 10/ neutrons. The generator has been designed to be easily repaired and modified. The unit requires no additional equipment for operation or measurement of output.

  17. Identifying Activity Cliff Generators of PPAR Ligands Using SAS Maps.

    PubMed

    Méndez-Lucio, Oscar; Pérez-Villanueva, Jaime; Castillo, Rafael; Medina-Franco, José L

    2012-12-01

    Structure-activity relationships (SAR) of compound databases play a key role in hit identification and lead optimization. In particular, activity cliffs, defined as a pair of structurally similar molecules that present large changes in potency, provide valuable SAR information. Herein, we introduce the concept of activity cliff generator, defined as a molecular structure that has a high probability to form activity cliffs with molecules tested in the same biological assay. To illustrate this concept, we discuss a case study where Structure-Activity Similarity maps were used to systematically identify and analyze activity cliff generators present in a dataset of 168 compounds tested against three peroxisome-proliferator-activated receptor (PPAR) subtypes. Single-target and dual-target activity cliff generators for PPARα and δ were identified. In addition, docking calculations of compounds that were classified as cliff generators helped to suggest a hot spot in the target protein responsible of activity cliffs and to analyze its implication in ligand-enzyme interaction.

  18. Active Generations: An Intergenerational Approach to Preventing Childhood Obesity

    ERIC Educational Resources Information Center

    Werner, Danilea; Teufel, James; Holtgrave, Peter L.; Brown, Stephen L.

    2012-01-01

    Background: Over the last 3 decades, US obesity rates have increased dramatically as more children and more adults become obese. This study explores an innovative program, Active Generations, an intergenerational nutrition education and activity program implemented in out-of-school environments (after school and summer camps). It utilizes older…

  19. An evolutionarily conserved NPC subcomplex, which redistributes in part to kinetochores in mammalian cells

    PubMed Central

    Belgareh, Naïma; Rabut, Gwénaël; Baï, Siau Wei; van Overbeek, Megan; Beaudouin, Joël; Daigle, Nathalie; Zatsepina, Olga V.; Pasteau, Fabien; Labas, Valérie; Fromont-Racine, Micheline; Ellenberg, Jan; Doye, Valérie

    2001-01-01

    The nuclear pore complexes (NPCs) are evolutionarily conserved assemblies that allow traffic between the cytoplasm and the nucleus. In this study, we have identified and characterized a novel human nuclear pore protein, hNup133, through its homology with the Saccharomyces cerevisiae nucleoporin scNup133. Two-hybrid screens and immunoprecipitation experiments revealed a direct and evolutionarily conserved interaction between Nup133 and Nup84/Nup107 and indicated that hNup133 and hNup107 are part of a NPC subcomplex that contains two other nucleoporins (the previously characterized hNup96 and a novel nucleoporin designated as hNup120) homologous to constituents of the scNup84 subcomplex. We further demonstrate that hNup133 and hNup107 are localized on both sides of the NPC to which they are stably associated at interphase, remain associated as part of a NPC subcomplex during mitosis, and are targeted at early stages to the reforming nuclear envelope. Throughout mitosis, a fraction of hNup133 and hNup107 localizes to the kinetochores, thus revealing an unexpected connection between structural NPCs constituents and kinetochores. Photobleaching experiments further showed that the mitotic cytoplasm contains kinetochore-binding competent hNup133 molecules and that in contrast to its stable association with the NPCs the interaction of this nucleoporin with kinetochores is dynamic. PMID:11564755

  20. omicsNPC: Applying the Non-Parametric Combination Methodology to the Integrative Analysis of Heterogeneous Omics Data

    PubMed Central

    Karathanasis, Nestoras; Tsamardinos, Ioannis

    2016-01-01

    Background The advance of omics technologies has made possible to measure several data modalities on a system of interest. In this work, we illustrate how the Non-Parametric Combination methodology, namely NPC, can be used for simultaneously assessing the association of different molecular quantities with an outcome of interest. We argue that NPC methods have several potential applications in integrating heterogeneous omics technologies, as for example identifying genes whose methylation and transcriptional levels are jointly deregulated, or finding proteins whose abundance shows the same trends of the expression of their encoding genes. Results We implemented the NPC methodology within “omicsNPC”, an R function specifically tailored for the characteristics of omics data. We compare omicsNPC against a range of alternative methods on simulated as well as on real data. Comparisons on simulated data point out that omicsNPC produces unbiased / calibrated p-values and performs equally or significantly better than the other methods included in the study; furthermore, the analysis of real data show that omicsNPC (a) exhibits higher statistical power than other methods, (b) it is easily applicable in a number of different scenarios, and (c) its results have improved biological interpretability. Conclusions The omicsNPC function competitively behaves in all comparisons conducted in this study. Taking into account that the method (i) requires minimal assumptions, (ii) it can be used on different studies designs and (iii) it captures the dependences among heterogeneous data modalities, omicsNPC provides a flexible and statistically powerful solution for the integrative analysis of different omics data. PMID:27812137

  1. Genetic variation at the NPC1L1 gene locus, plasma lipoproteins, and heart disease risk in the elderly

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Niemann-Pick C1-like 1 protein (NPC1L1) plays a critical role in intestinal cholesterol absorption. Our objective was to examine whether five variants (-133A>G, -18A>C, L272L, V1296V, and U3_28650A>G) at the NPC1L1 gene have effects on lipid levels, prevalence, and incidence of coronary heart diseas...

  2. "Paradox of slow frequencies" - Are slow frequencies in upper cortical layers a neural predisposition of the level/state of consciousness (NPC)?

    PubMed

    Northoff, Georg

    2017-04-05

    Consciousness research has much focused on faster frequencies like alpha or gamma while neglecting the slower ones in the infraslow (0.001-0.1Hz) and slow (0.1-1Hz) frequency range. These slower frequency ranges have a "bad reputation" though; their increase in power can observed during the loss of consciousness as in sleep, anesthesia, and vegetative state. However, at the same time, slower frequencies have been conceived instrumental for consciousness. The present paper aims to resolve this paradox which I describe as "paradox of slow frequencies". I first show various data that suggest a central role of slower frequencies in integrating faster ones, i.e., "temporo-spatial integration and nestedness". Such "temporo-spatial integration and nestedness" is disrupted during the loss of consciousness as in anesthesia and sleep leading to "temporo-spatial fragmentation and isolation" between slow and fast frequencies. Slow frequencies are supposedly mediated by neural activity in upper cortical layers in higher-order associative regions as distinguished from lower cortical layers that are related to faster frequencies. Taken together, slower and faster frequencies take on different roles for the level/state of consciousness. Faster frequencies by themselves are sufficient and thus a neural correlate of consciousness (NCC) while slower frequencies are a necessary non-sufficient condition of possible consciousness, e.g., a neural predisposition of the level/state of consciousness (NPC). This resolves the "paradox of slow frequencies" in that it assigns different roles to slower and faster frequencies in consciousness, i.e., NCC and NPC. Taken as NCC and NPC, fast and slow frequencies including their relation as in "temporo-spatial integration and nestedness" can be considered a first "building bloc" of a future "temporo-spatial theory of consciousness" (TTC) (Northoff, 2013; Northoff, 2014b; Northoff & Huang, 2017).

  3. Generator of chemically active low-temperature plasma

    NASA Astrophysics Data System (ADS)

    Tyuftyaev, A. S.; Gadzhiev, M. Kh; Sargsyan, M. A.; Demirov, N. A.; Spector, N. O.

    2016-11-01

    A new generator of high enthalpy (H 0 > 40 kJ/g), chemically active nitrogen and air plasmas was designed and constructed. Main feature of the generator is an expanding channel of an output electrode; the generator belongs to the class of DC plasma torches with thermionic cathode with an efficiency of 80%. The generator ensures the formation of a slightly divergent plasma jet (2α = 12°) with a diameter of D = 10-12 mm, an electric arc maximum power of 20-50 kW, plasma forming gas flow rate 1.0-2.0 g/s, and the average plasma temperature at an outlet of 8000-11000 K.

  4. Relationship between peptide membrane curvature generation and bactericidal activities

    NASA Astrophysics Data System (ADS)

    Schmidt, Nathan; Lee, Michelle; Kuo, David; Ouellette, Andre; Wong, Gerard

    2013-03-01

    Many amphipathic peptides and amphipathic domains in proteins can restructure biological membranes. Two examples are host defense antimicrobial peptides (AMPs) which disrupt and destabilize the cell membranes of microbes, and apolipoproteins which help stabilize nanoscale lipid aggregates. We use complementary x-ray and bacterial cell assays to elucidate the molecular length scale membrane deformations generated by amphipathic peptides with different structural motifs and relate these deformations to their activities on bacteria. Small angle x-ray scattering is used to study the interactions of model membranes with prototypical AMPs and consensus peptides from the amphipathic domains in apolipoproteins. By characterizing the nanoscale curvature deformations induced by these two distinct classes of membrane restructuring peptides we will discuss the role of amino acid composition on curvature generation. Bactericidal assays are used to access the in vivo activities of different amphipathic peptide motifs in order to understand the relationships between cell viability and membrane curvature generation.

  5. Income Generation Activities among Academic Staffs at Malaysian Public Universities

    ERIC Educational Resources Information Center

    Ahmad, Abd Rahman; Soon, Ng Kim; Ting, Ngeoh Pei

    2015-01-01

    Income generation activities have been acquainted among public higher education institutions (HEIs) in Malaysia. Various factors that brought to insufficient of funding caused Higher Education Institutions(HEIs) to seek for additional income as to support the operation expenses. Financial sustainability issues made up the significant impact…

  6. Density of Plutonium Turnings Generated from Machining Activities

    SciTech Connect

    Gonzales, John Robert; Vigil, Duane M.; Jachimowski, Thomas A.; Archuleta, Alonso; Arellano, Gerald Joseph; Melton, Vince Lee

    2016-10-20

    The purpose of this project was to determine the density of plutonium (Pu) turnings generated from the range of machining activities, using both surrogate material and machined Pu turnings. Verify that 500 grams (g) of plutonium will fit in a one quart container using a surrogate equivalent volume and that 100 grams of Pu will fit in a one quart Savy container.

  7. Entanglement generation between unstable optically active qubits without photodetectors

    SciTech Connect

    Matsuzaki, Yuichiro; Solinas, Paolo

    2011-09-15

    We propose a robust deterministic scheme to generate entanglement at high fidelity without the need for photodetectors even for quantum bits (qubits) with extremely poor optically active states. Our protocol employs stimulated Raman adiabatic passage for population transfer without actually exciting the system. Furthermore, it is found to be effective even if the environmental decoherence rate is of the same order of magnitude as the atom-photon coupling frequency. Our scheme has the potential to solve entanglement generation problems, e.g., in distributed quantum computing.

  8. Relationship between short activated partial thromboplastin times, thrombin generation, procoagulant factors and procoagulant phospholipid activity.

    PubMed

    Mina, Ashraf; Favaloro, Emmanuel J; Koutts, Jerry

    2012-04-01

    Short activated partial thromboplastin times (APTTs) are associated with thrombosis. However, what short APTTs actually represent in terms of possible mechanistic pathways is not well characterized. We have assessed thrombin generation as compared with levels of procoagulant factor (fibrinogen, V, VIII, IX, XI and XII) activities, von Willebrand factor level and activity using collagen binding, as well as procoagulant phospholipid activity, in 113 consecutive samples exhibiting a short APTT compared with an equal number of age-matched and sex-matched samples yielding a normal APTT. We found a significant difference in peak thrombin generation, velocity index and area under the curve between the two groups, and that thrombin generation markers correlated with the APTT, procoagulant phospholipid activity and several procoagulant clotting factors. We conclude that short APTTs represent a procoagulant milieu, as represented by heightened thrombin generation and several other heightened procoagulant activities, which may help explain the association with thrombosis.

  9. The role of Niemann-Pick C1 - Like 1 (NPC1L1) in intestinal sterol absorption.

    PubMed

    Turley, Stephen D

    2008-04-01

    The absorption of cholesterol by the proximal small intestine represents a major pathway for the entry of cholesterol into the body pools. This cholesterol is derived primarily from the bile and the diet. In adult humans, typically several hundred milligrams of cholesterol reach the liver from the intestine daily, with the potential to impact the plasma low density lipoprotein-cholesterol (LDL-C) concentration. There are three main phases involved in cholesterol absorption. The first occurs intraluminally and culminates in micellar solubilization of unesterified cholesterol which facilitates its movement up to the brush border membrane (BBM) of the enterocyte. The second phase involves the transport of cholesterol across the BBM by Niemann-Pick C1 Like-1 (NPC1L1), while the third phase entails a series of steps within the enterocyte involving the esterification of cholesterol and its incorporation, along with other lipids and apolipoprotein B48 (apo B48), into nascent chylomicrons (CM). The discovery of the role of NPC1L1 in intestinal sterol transport occurred directly as a consequence of efforts to identify the molecular target of ezetimibe, a novel, potent, and specific inhibitor of sterol absorption that is now widely used in combination therapy with statins for the management of hypercholesterolemia in the general population. Some aspects of the role of NPC1L1 in cholesterol absorption nevertheless remain controversial and are the subject of ongoing research. For example, one report suggests that NPC1L1 is located not in the plasma membrane but intracellularly where it is thought to be involved in cytosolic trafficking of cholesterol, while another concludes that a protein other than NPC1L1 is responsible for the high affinity binding of cholesterol on intestinal BBM. However, other new studies which show that the in vivo responsiveness of different species to ezetimibe correlates with NPC1L1 binding affinity further support the widely held belief that NPC1L1

  10. Orlistat limits cholesterol intestinal absorption by Niemann-pick C1-like 1 (NPC1L1) inhibition.

    PubMed

    Alqahtani, Saeed; Qosa, Hisham; Primeaux, Brian; Kaddoumi, Amal

    2015-09-05

    The known mechanism by which orlistat decreases the absorption of dietary cholesterol is by inhibition of intestinal lipases. The aim of this study was to investigate the ability of orlistat to limit cholesterol absorption by inhibition of the cholesterol transport protein Niemann-Pick C1-like 1 (NPC1L1) as another mechanism of action. In situ rat intestinal perfusion studies were conducted to study the effect of orlistat on jejunal cholesterol absorption. Inhibition kinetic parameters were calculated from in vitro inhibition studies using Caco2 and NPC1L1 transfected cell lines. The in situ studies demonstrated that intestinal perfusion of orlistat (100µM) was able to reduce cholesterol absorption by three-fold when compared to control (i.e. in the absence of orlistat, P<0.01). In vitro studies using Caco2 cells demonstrated orlistat to reduce the cellular uptake of cholesterol by 30%. Additionally, orlistat reduced the cellular uptake of cholesterol in dose dependent manner in NPC1L1 transfected cell line with an IC50=1.2µM. Lineweaver-Burk plot indicated a noncompetitive inhibition of NPC1L1 by orlistat. Beside the already established mechanism by which orlistat reduces the absorption of cholesterol, we demonstrated for the first time that orlistat limits cholesterol absorption by the inhibition of NPC1L1 transport protein.

  11. Income-generating activities for family planning acceptors.

    PubMed

    1989-07-01

    The Income Generating Activities program for Family Planning Acceptors was introduced in Indonesia in 1979. Capital input by the Indonesian National Family Planning Coordination Board and the UN Fund for Population Activities was used to set up small businesses by family planning acceptors. In 2 years, when the businesses become self-sufficient, the loans are repaid, and the money is used to set up new family planning acceptors in business. The program strengthens family planning acceptance, improves the status of women, and enhances community self-reliance. The increase in household income generated by the program raises the standards of child nutrition, encourages reliance on the survival of children, and decreases the value of large families. Approximately 18,000 Family Planning-Income Generating Activities groups are now functioning all over Indonesia, with financial assistance from the central and local governments, the World Bank, the US Agency for International Development, the UN Population Fund, the Government of the Netherlands, and the Government of Australia through the Association of South East Asian Nations.

  12. Enhanced ionospheric plasma bubble generation in more active ITCZ

    NASA Astrophysics Data System (ADS)

    Li, Guozhu; Otsuka, Yuichi; Ning, Baiqi; Abdu, M. A.; Yamamoto, M.; Wan, Weixing; Liu, Libo; Abadi, Prayitno

    2016-03-01

    A close link between the atmospheric Intertropical Convergence Zone (ITCZ) and ionospheric plasma bubble has been proposed since the last century. But this relationship has often appeared to be less than convincing due to the simultaneous roles played by several other factors in shaping the global distribution of ionospheric bubbles. From simultaneous collaborative radar multibeam steering measurements at Kototabang (0.2°S, 100.3°E) and Sanya (18.4°N, 109.6°E), conducted during September-October of 2012 and 2013, we find that the total numbers of nights with bubble (i.e., occurrence rates) at the two closely located longitudes (Kototabang and Sanya) are comparable. But interestingly, the total number of nights with locally generated bubble (i.e., generation rate) over Kototabang is clearly more than that over Sanya. Further analysis reveals that a more active ITCZ is situated around the longitude of Kototabang. We surmise that the enhanced ionospheric bubble generation at Kototabang longitude could be caused by a higher gravity wave activity associated with the more active ITCZ.

  13. Npc1 deficiency in the C57BL/6J genetic background enhances Niemann-Pick disease type C spleen pathology.

    PubMed

    Parra, Julio; Klein, Andrés D; Castro, Juan; Morales, María Gabriela; Mosqueira, Matías; Valencia, Ilse; Cortés, Victor; Rigotti, Attilio; Zanlungo, Silvana

    2011-09-30

    Niemann-Pick type C (NPC) disease is an autosomal recessive neurovisceral lipid storage disorder. The affected genes are NPC1 and NPC2. Mutations in either gene lead to intracellular cholesterol accumulation. There are three forms of the disease, which are categorized based on the onset and severity of the disease: the infantile form, in which the liver and spleen are severely affected, the juvenile form, in which the liver and brain are affected, and the adult form, which affects the brain. In mice, a spontaneous mutation in the Npc1 gene originated in the BALB/c inbred strain mimics the juvenile form of the disease. To study the influence of genetic background on the expression of NPC disease in mice, we transferred the Npc1 mutation from the BALB/c to C57BL/6J inbred background. We found that C57BL/6J-Npc1(-/-) mice present with a much more aggressive form of the disease, including a shorter lifespan than BALB/c-Npc1(-/-) mice. Surprisingly, there was no difference in the amount of cholesterol in the brains of Npc1(-/-) mice of either mouse strain. However, Npc1(-/-) mice with the C57BL/6J genetic background showed striking spleen damage with a marked buildup of cholesterol and phospholipids at an early age, which correlated with large foamy cell clusters. In addition, C57BL/6J Npc1(-/-) mice presented red cell abnormalities and abundant ghost erythrocytes that correlated with a lower hemoglobin concentration. We also found abnormalities in white cells, such as cytoplasmic granulation and neutrophil hypersegmentation that included lymphopenia and atypias. In conclusion, Npc1 deficiency in the C57BL6/J background is associated with spleen, erythrocyte, and immune system abnormalities that lead to a reduced lifespan.

  14. Snubber-less NPC inverter by a novel reduction technique of parasitic inductance for magnet power supplies

    NASA Astrophysics Data System (ADS)

    Koseki, K.; Morita, Y.

    2014-10-01

    The effects induced by parasitic inductance were studied in a neutral point clamped (NPC) inverter. The energy stored in the parasitic inductance by the output current causes a surge voltage during the turn-off period of semiconductor switches. The effect is serious in a magnet power supply for which a large excitation current is required. It is predicted by a circuit analysis that the parasitic inductance causes an electrical breakdown of the semiconductor switches. It was found that the most promising way to mitigate the effect is to reduce the parasitic inductance. With newly developed circuitry and layout of an NPC inverter, cancelation of the induced magnetic field, which is based on Amperes circuital law, in power devices has been accomplished. The newly developed NPC inverter has been operated successfully with both a resistive dummy load and the dipole magnets in the main ring synchrotron at the J-PARC facility.

  15. Atmospheric wind field conditions generated by active grids

    NASA Astrophysics Data System (ADS)

    Knebel, Pascal; Kittel, Achim; Peinke, Joachim

    2011-08-01

    An active grid for turbulence generation of several rotatable axes with surmounted vanes that can be driven via stepper or servo motors is presented. We investigate the impact of different excitation protocols for the grid. Using such protocols that already have the intermittent structure of turbulence, higher intermittent flows can be achieved. This concept can also be used to generate turbulent flows of high turbulence intensities (>25%) exhibiting integral length scales beyond the typical size of the test section of the wind tunnel. Similar two-point correlations measured by the intermittent statistics of velocity increments that are characteristic for flows of high Reynolds number, i.e. in the atmospheric boundary layer, can be reproduced.

  16. Characterization of a Spontaneous Novel Mutation in the NPC2 Gene in a Cat Affected by Niemann Pick Type C Disease

    PubMed Central

    Zampieri, Stefania; Bianchi, Ezio; Cantile, Carlo; Saleri, Roberta; Bembi, Bruno; Dardis, Andrea

    2014-01-01

    Niemann-Pick C disease (NPC) is an autosomal recessive lysosomal storage disorder characterized by accumulation of unesterified cholesterol and other lipids within the lysosomes due to mutation in NPC1 or NPC2 genes. A feline model of NPC carrying a mutation in NPC1 gene has been previously described. We have identified two kittens affected by NPC disease due to a mutation in NPC2 gene. They manifested with tremors at the age of 3 months, which progressed to dystonia and severe ataxia. At 6 months of age cat 2 was unable to stand without assistance and had bilaterally reduced menace response. It died at the age of 10 months. Post-mortem histological analysis of the brain showed the presence of neurons with cytoplasmic swelling and vacuoles, gliosis of the substantia nigra and degeneration of the white matter. Spheroids with accumulation of ubiquitinated aggregates were prominent in the cerebellar cortex. Purkinje cells were markedly reduced in number and they showed prominent intracytoplasmic storage. Scattered perivascular aggregates of lymphocytes and microglial cells proliferation were present in the thalamus and midbrain. Proliferation of Bergmann glia was also observed. In the liver, hepatocytes were swollen because of accumulation of small vacuoles and foamy Kupffer cells were also detected. Foamy macrophages were observed within the pulmonary interstitium and alveoli as well. At 9 months cat 1 was unable to walk, developed seizures and it was euthanized at 21 months. Filipin staining of cultured fibroblasts showed massive storage of unesterified cholesterol. Molecular analysis of NPC1 and NPC2 genes showed the presence of a homozygous intronic mutation (c.82+5G>A) in the NPC2 gene. The subsequent analysis of the mRNA showed that the mutation causes the retention of 105 bp in the mature mRNA, which leads to the in frame insertion of 35 amino acids between residues 28 and 29 of NPC2 protein (p.G28_S29ins35). PMID:25396745

  17. Studies on antistreptolysin O activity generated in serum by microorganisms.

    PubMed Central

    Watson, K. C.; Kerr, E. J.

    1976-01-01

    An antistreptolysin factor (ASF) was generated in normal human serum by the growth of Staph, aureus and Pseud, aeruginosa. Alpha toxin producing strains of the former were usually positive but activity was not restricted to such strains. Positive strains produce cholesterol esterase which was obtained from DEAE-cellulose column fractions of 18 h broth cultures. Antistreptolysin factor develops slowly in serum, being maximal between the 5th and 10th days and is associated with alterations and disappearance of beta lipoproteins on gel electrophoresis. Activity also appeared in beta lipoproteins precipitated from normal serum with dextran sulphate and redissolved in nutrient broth before inoculation with Staph, aureus. The slow appearance of antistreptolysin activity in serum appears to be due to an esterase inhibitor which is present in high concentrations in some sera. Activity is also modified by the production of a staphylococcal fraction capable of binding to the antistreptolysin factor and reducing its activity. It is suggested that antistreptolysin factor which can be demonstrated in small amounts in normal human serum represents a readily available non-specific defence mechanism capable of binding to certain bacterial products and possible to other foreign protein molecules. Images Fig. 2 PMID:178333

  18. A marked paucity of granule cells in the developing cerebellum of the Npc1−/− mouse is corrected by a single injection of hydroxypropyl-β-cyclodextrin

    PubMed Central

    Nusca, S.; Canterini, S.; Palladino, G.; Bruno, F.; Mangia, F.; Erickson, R.P.; Fiorenza, M.T.

    2014-01-01

    In this study we show that postnatal development of cerebellar granule neurons (GNs) is defective in Npc1−/− mice. Compared to age-matched wild-type littermates, there is an accelerated disappearance of the external granule layer (EGL) in these mice. This is due to a premature exit from the cell cycle of GN precursors residing at the level of the EGL. As a consequence, the size of cerebellar lobules of these mice displays a 20%–25% reduction compared to that of age-matched wild-type mice. This size reduction is detectable at post-natal day 28 (PN28), when cerebellar GN development is completed while signs of neuronal atrophy are not yet apparent. Based on the analysis of EGL thickness and the determination of proliferating GN fractions at increasing developmental times (PN8–PN14), we trace the onset of this GN developmental defect during the second postnatal week. We also show that during this developmental time Shh transcripts undergo a significant reduction in Npc1−/− mice compared to age-matched wild-type mice. In light of the mitogenic activity of Shh on GNs, this observation further supports the presence of defective GN proliferation in Npc1−/− mice. A single injection of hydroxypropyl-β-cyclodextrin at PN7 rescues this defect, restoring the normal patterns of granule neuron proliferation and cerebellar lobule size. To our knowledge, these findings identify a novel developmental defect that was underappreciated in previous studies. This defect was probably overlooked because Npc1 loss-of-function does not affect cerebellar foliation and causes the internal granule layer and molecular layer to decrease proportionally, giving rise to a normally appearing, yet harmoniously smaller, cerebellum. PMID:24969023

  19. An exploratory propensity score matched comparison of second-generation and first-generation baroreflex activation therapy systems.

    PubMed

    Wachter, Rolf; Halbach, Marcel; Bakris, George L; Bisognano, John D; Haller, Hermann; Beige, Joachim; Kroon, Abraham A; Nadim, Mitra K; Lovett, Eric G; Schafer, Jill E; de Leeuw, Peter W

    2016-12-16

    Baroreflex activation therapy (BAT) is a device-based therapy for patients with treatment-resistant hypertension. In a randomized, controlled trial, the first-generation system significantly reduced blood pressure (BP) versus sham. Although an open-label validation study of the second-generation system demonstrated similar BP reductions, controlled data are not presently available. Therefore, this investigation compares results of first- and second-generation BAT systems. Two cohorts of first-generation BAT system patients were generated with propensity matching to compare against the validation group of 30 second-generation subjects. The first cohort was drawn from the first-generation randomized trial sham group and the second cohort from the active therapy group. Safety and efficacy were compared for the second-generation group relative to the first generation. At 6 months, second-generation BAT outperformed first-generation sham systolic BP reduction by 20 ± 28 mm Hg (mean ± standard deviation, P = .008), while BP reduction in first- and second-generation active groups was similar. At 12 months, efficacy was comparable between all three groups after the sham group had received 6 months of therapy; 47% of second-generation patients achieved goal systolic BP of 140 mm Hg or less after 12 months, comparable to 50% of patients at goal in the first-generation group (P > .999). Implant procedure time, system/procedural safety, and pulse generator longevity improved with the second-generation system. Propensity-matched cohort analysis of the first- and second-generation BAT systems suggests similar therapeutic benefit and superior BP reduction of the second-generation system relative to sham control. Implantation procedure duration and perioperative safety were improved with the second-generation device. These findings should be validated in a prospective randomized trial.

  20. Active control of fan-generated plane wave noise

    NASA Astrophysics Data System (ADS)

    Gerhold, Carl H.; Nuckolls, William E.; Santamaria, Odillyn L.; Martinson, Scott D.

    1993-08-01

    Subsonic propulsion systems for future aircraft may incorporate ultra-high bypass ratio ducted fan engines whose dominant noise source is the fan with blade passage frequency less than 1000 Hz. This low frequency combines with the requirement of a short nacelle to diminish the effectiveness of passive duct liners. Active noise control is seen as a viable method to augment the conventional passive treatments. An experiment to control ducted fan noise using a time domain active adaptive system is reported. The control sound source consists of loudspeakers arrayed around the fan duct. The error sensor location is in the fan duct. The purpose of this experiment is to demonstrate that the in-duct error sensor reduces the mode spillover in the far field, thereby increasing the efficiency of the control system. In this first series of tests, the fan is configured so that predominantly zero order circumferential waves are generated. The control system is found to reduce the blade passage frequency tone significantly in the acoustic far field when the mode orders of the noise source and of the control source are the same. The noise reduction is not as great when the mode orders are not the same even though the noise source modes are evanescent, but the control system converges stably and global noise reduction is demonstrated in the far field. Further experimentation is planned in which the performance of the system will be evaluated when higher order radial and spinning modes are generated.

  1. Experimental generation of single photons via active multiplexing

    SciTech Connect

    Ma Xiaosong; Zotter, Stefan; Kofler, Johannes; Jennewein, Thomas; Zeilinger, Anton

    2011-04-15

    An on-demand single-photon source is a fundamental building block in quantum science and technology. We experimentally demonstrate the proof of concept for a scheme to generate on-demand single photons via actively multiplexing several heralded photons probabilistically produced from pulsed spontaneous parametric down-conversions (SPDCs). By utilizing a four-photon-pair source, an active feed-forward technique, and an ultrafast single-photon router, we show a fourfold enhancement of the output photon rate. Simultaneously, we maintain the quality of the output single-photon states, confirmed by correlation measurements. We also experimentally verify, via Hong-Ou-Mandel interference, that the router does not affect the indistinguishability of the single photons. Furthermore, we give numerical simulations, which indicate that photons based on multiplexing of four SPDC sources can outperform the heralding based on highly advanced photon-number-resolving detectors. Our results show a route for on-demand single-photon generation and the practical realization of scalable linear optical quantum-information processing.

  2. Active control of fan-generated plane wave noise

    NASA Technical Reports Server (NTRS)

    Gerhold, Carl H.; Nuckolls, William E.; Santamaria, Odillyn L.; Martinson, Scott D.

    1993-01-01

    Subsonic propulsion systems for future aircraft may incorporate ultra-high bypass ratio ducted fan engines whose dominant noise source is the fan with blade passage frequency less than 1000 Hz. This low frequency combines with the requirement of a short nacelle to diminish the effectiveness of passive duct liners. Active noise control is seen as a viable method to augment the conventional passive treatments. An experiment to control ducted fan noise using a time domain active adaptive system is reported. The control sound source consists of loudspeakers arrayed around the fan duct. The error sensor location is in the fan duct. The purpose of this experiment is to demonstrate that the in-duct error sensor reduces the mode spillover in the far field, thereby increasing the efficiency of the control system. In this first series of tests, the fan is configured so that predominantly zero order circumferential waves are generated. The control system is found to reduce the blade passage frequency tone significantly in the acoustic far field when the mode orders of the noise source and of the control source are the same. The noise reduction is not as great when the mode orders are not the same even though the noise source modes are evanescent, but the control system converges stably and global noise reduction is demonstrated in the far field. Further experimentation is planned in which the performance of the system will be evaluated when higher order radial and spinning modes are generated.

  3. Contact system activation and high thrombin generation in hyperthyroidism.

    PubMed

    Kim, Namhee; Gu, Ja-Yoon; Yoo, Hyun Ju; Han, Se Eun; Kim, Young Il; Nam-Goong, Il Seong; Kim, Eun Sook; Kim, Hyun Kyung

    2017-01-30

    Background Hyperthyroidism is associated with increased thrombotic risk. Since contact system activation through formation of neutrophil extracellular traps (NET) has emerged as an important trigger of thrombosis, we hypothesized that the contact system is activated along with active NET formation in hyperthyroidism and that their markers correlate with disease severity. Materials and Methods In 61 patients with hyperthyroidism and 40 normal controls, the levels of coagulation factors (fibrinogen, and factor VII, VIII, IX, XI, and XII), D-dimer, thrombin generation assay (TGA) markers, NET formation markers (histone-DNA complex, double-stranded DNA, neutrophil elastase), and contact system markers (activated factor XII [XIIa], high-molecular-weight kininogen [HMWK], prekallikrein, and bradykinin) were measured. Results Patients with hyperthyroidism showed higher levels of fibrinogen [median (interquartile range), 315 (280-344) versus 262 (223-300), P=0.001], D-dimer [103.8 (64.8-151.5) versus 50.7 (37.4-76.0), P<0.001], peak thrombin [131.9 (102.2-159.4) versus 31.6 (14.8-83.7), P<0.001] and endogenous thrombin potential [649 (538-736) versus 367 (197-1147), P=0.021] in TGA with 1 pM tissue factor, neutrophil elastase [1.10 (0.39-2.18) versus 0.23 (0.20-0.35), P<0.001], factor XIIa [66.9 (52.8-87.0) versus 73.0 (57.1-86.6), P<0.001], HMWK [6.11 (4.95-7.98) versus 3.83 (2.60-5.68), P<0.001], prekallikrein [2.15 (1.00-6.36) versus 1.41 (0.63-2.22), P=0.026], and bradykinin [152.4 (137.6-180.4) versus 118.3 (97.1-137.9), P<0.001] than did normal controls. In age- and sex-adjusted logistic regression analysis, fibrinogen, factor VIII, IX, and XIIa, D-dimer, peak thrombin, neutrophil elastase, HMWK, and bradykinin showed significant odds ratios representing hyperthyroidism's contribution to coagulation and contact system activation. Free T4 was significantly correlated with factors VIII and IX, D-dimer, double-stranded DNA, and bradykinin. Conclusion This study

  4. Connection between solar activity cycles and grand minima generation

    NASA Astrophysics Data System (ADS)

    Vecchio, A.; Lepreti, F.; Laurenza, M.; Alberti, T.; Carbone, V.

    2017-03-01

    Aims: The revised dataset of sunspot and group numbers (released by WDC-SILSO) and the sunspot number reconstruction based on dendrochronologically dated radiocarbon concentrations have been analyzed to provide a deeper characterization of the solar activity main periodicities and to investigate the role of the Gleissberg and Suess cycles in the grand minima occurrence. Methods: Empirical mode decomposition (EMD) has been used to isolate the time behavior of the different solar activity periodicities. A general consistency among the results from all the analyzed datasets verifies the reliability of the EMD approach. Results: The analysis on the revised sunspot data indicates that the highest energy content is associated with the Schwabe cycle. In correspondence with the grand minima (Maunder and Dalton), the frequency of this cycle changes to longer timescales of 14 yr. The Gleissberg and Suess cycles, with timescales of 60-120 yr and 200-300 yr, respectively, represent the most energetic contribution to sunspot number reconstruction records and are both found to be characterized by multiple scales of oscillation. The grand minima generation and the origin of the two expected distinct types of grand minima, Maunder and longer Spörer-like, are naturally explained through the EMD approach. We found that the grand minima sequence is produced by the coupling between Gleissberg and Suess cycles, the latter being responsible for the most intense and longest Spörer-like minima (with typical duration longer than 80 yr). Finally, we identified a non-solar component, characterized by a very long scale oscillation of 7000 yr, and the Hallstatt cycle ( 2000 yr), likely due to the solar activity. Conclusions: These results provide new observational constraints on the properties of the solar cycle periodicities, the grand minima generation, and thus the long-term behavior of the solar dynamo.

  5. A novel approach to analyze lysosomal dysfunctions through subcellular proteomics and lipidomics: the case of NPC1 deficiency

    NASA Astrophysics Data System (ADS)

    Tharkeshwar, Arun Kumar; Trekker, Jesse; Vermeire, Wendy; Pauwels, Jarne; Sannerud, Ragna; Priestman, David A.; Te Vruchte, Danielle; Vints, Katlijn; Baatsen, Pieter; Decuypere, Jean-Paul; Lu, Huiqi; Martin, Shaun; Vangheluwe, Peter; Swinnen, Johannes V.; Lagae, Liesbet; Impens, Francis; Platt, Frances M.; Gevaert, Kris; Annaert, Wim

    2017-01-01

    Superparamagnetic iron oxide nanoparticles (SPIONs) have mainly been used as cellular carriers for genes and therapeutic products, while their use in subcellular organelle isolation remains underexploited. We engineered SPIONs targeting distinct subcellular compartments. Dimercaptosuccinic acid-coated SPIONs are internalized and accumulate in late endosomes/lysosomes, while aminolipid-SPIONs reside at the plasma membrane. These features allowed us to establish standardized magnetic isolation procedures for these membrane compartments with a yield and purity permitting proteomic and lipidomic profiling. We validated our approach by comparing the biomolecular compositions of lysosomes and plasma membranes isolated from wild-type and Niemann-Pick disease type C1 (NPC1) deficient cells. While the accumulation of cholesterol and glycosphingolipids is seen as a primary hallmark of NPC1 deficiency, our lipidomics analysis revealed the buildup of several species of glycerophospholipids and other storage lipids in selectively late endosomes/lysosomes of NPC1-KO cells. While the plasma membrane proteome remained largely invariable, we observed pronounced alterations in several proteins linked to autophagy and lysosomal catabolism reflecting vesicular transport obstruction and defective lysosomal turnover resulting from NPC1 deficiency. Thus the use of SPIONs provides a major advancement in fingerprinting subcellular compartments, with an increased potential to identify disease-related alterations in their biomolecular compositions.

  6. A novel approach to analyze lysosomal dysfunctions through subcellular proteomics and lipidomics: the case of NPC1 deficiency

    PubMed Central

    Tharkeshwar, Arun Kumar; Trekker, Jesse; Vermeire, Wendy; Pauwels, Jarne; Sannerud, Ragna; Priestman, David A.; te Vruchte, Danielle; Vints, Katlijn; Baatsen, Pieter; Decuypere, Jean-Paul; Lu, Huiqi; Martin, Shaun; Vangheluwe, Peter; Swinnen, Johannes V.; Lagae, Liesbet; Impens, Francis; Platt, Frances M.; Gevaert, Kris; Annaert, Wim

    2017-01-01

    Superparamagnetic iron oxide nanoparticles (SPIONs) have mainly been used as cellular carriers for genes and therapeutic products, while their use in subcellular organelle isolation remains underexploited. We engineered SPIONs targeting distinct subcellular compartments. Dimercaptosuccinic acid-coated SPIONs are internalized and accumulate in late endosomes/lysosomes, while aminolipid-SPIONs reside at the plasma membrane. These features allowed us to establish standardized magnetic isolation procedures for these membrane compartments with a yield and purity permitting proteomic and lipidomic profiling. We validated our approach by comparing the biomolecular compositions of lysosomes and plasma membranes isolated from wild-type and Niemann-Pick disease type C1 (NPC1) deficient cells. While the accumulation of cholesterol and glycosphingolipids is seen as a primary hallmark of NPC1 deficiency, our lipidomics analysis revealed the buildup of several species of glycerophospholipids and other storage lipids in selectively late endosomes/lysosomes of NPC1-KO cells. While the plasma membrane proteome remained largely invariable, we observed pronounced alterations in several proteins linked to autophagy and lysosomal catabolism reflecting vesicular transport obstruction and defective lysosomal turnover resulting from NPC1 deficiency. Thus the use of SPIONs provides a major advancement in fingerprinting subcellular compartments, with an increased potential to identify disease-related alterations in their biomolecular compositions. PMID:28134274

  7. A novel approach to analyze lysosomal dysfunctions through subcellular proteomics and lipidomics: the case of NPC1 deficiency.

    PubMed

    Tharkeshwar, Arun Kumar; Trekker, Jesse; Vermeire, Wendy; Pauwels, Jarne; Sannerud, Ragna; Priestman, David A; Te Vruchte, Danielle; Vints, Katlijn; Baatsen, Pieter; Decuypere, Jean-Paul; Lu, Huiqi; Martin, Shaun; Vangheluwe, Peter; Swinnen, Johannes V; Lagae, Liesbet; Impens, Francis; Platt, Frances M; Gevaert, Kris; Annaert, Wim

    2017-01-30

    Superparamagnetic iron oxide nanoparticles (SPIONs) have mainly been used as cellular carriers for genes and therapeutic products, while their use in subcellular organelle isolation remains underexploited. We engineered SPIONs targeting distinct subcellular compartments. Dimercaptosuccinic acid-coated SPIONs are internalized and accumulate in late endosomes/lysosomes, while aminolipid-SPIONs reside at the plasma membrane. These features allowed us to establish standardized magnetic isolation procedures for these membrane compartments with a yield and purity permitting proteomic and lipidomic profiling. We validated our approach by comparing the biomolecular compositions of lysosomes and plasma membranes isolated from wild-type and Niemann-Pick disease type C1 (NPC1) deficient cells. While the accumulation of cholesterol and glycosphingolipids is seen as a primary hallmark of NPC1 deficiency, our lipidomics analysis revealed the buildup of several species of glycerophospholipids and other storage lipids in selectively late endosomes/lysosomes of NPC1-KO cells. While the plasma membrane proteome remained largely invariable, we observed pronounced alterations in several proteins linked to autophagy and lysosomal catabolism reflecting vesicular transport obstruction and defective lysosomal turnover resulting from NPC1 deficiency. Thus the use of SPIONs provides a major advancement in fingerprinting subcellular compartments, with an increased potential to identify disease-related alterations in their biomolecular compositions.

  8. Enumerating Minimal Active Metabolic Pathways by Model Generation

    NASA Astrophysics Data System (ADS)

    Soh, Takehide; Inoue, Katsumi

    In systems biology, identifying vital functions like glycolysis from a given metabolic pathway is important to understand living organisms. In this paper, we particularly focus on the problem of enumerating minimal active pathways producing target metabolites from source metabolites. We represent the problem in propositional formulas and solve it through minimal model generation. An advantage of our method is that each solution satisfies qualitative laws of biochemical reactions. Moreover, we can calculate such solutions for a cellular scale metabolic pathway within a few seconds. In experiments, we have applied our method to a whole Escherichia coli metabolic pathway. As a result, we found a minimal set of reactions corresponding to the conventional glycolysis pathway described in a biological database EcoCyc.

  9. Microfluidic Platform Generates Oxygen Landscapes for Localized Hypoxic Activation

    PubMed Central

    Rexius, Megan L.; Mauleon, Gerardo; Malik, Asrar B.; Rehman, Jalees; Eddington, David T.

    2014-01-01

    An open-well microfluidic platform generates an oxygen landscape using gas-perfused networks which diffuse across a membrane. The device enables real-time analysis of cellular and tissue responses to oxygen tension to define how cells adapt to heterogeneous oxygen conditions found in the physiological setting. We demonstrate that localized hypoxic activation of cells elicited specific metabolic and gene responses in human microvascular endothelial cells and bone marrow-derived mesenchymal stem cells. A robust demonstration of the compatibility of the device with standard laboratory techniques demonstrates the wide utility of the method. This platform is ideally suited to study real-time cell responses and cell-cell interactions within physiologically relevant oxygen landscapes. PMID:25315003

  10. Microfluidic platform generates oxygen landscapes for localized hypoxic activation.

    PubMed

    Rexius-Hall, Megan L; Mauleon, Gerardo; Malik, Asrar B; Rehman, Jalees; Eddington, David T

    2014-12-21

    An open-well microfluidic platform generates an oxygen landscape using gas-perfused networks which diffuse across a membrane. The device enables real-time analysis of cellular and tissue responses to oxygen tension to define how cells adapt to heterogeneous oxygen conditions found in the physiological setting. We demonstrate that localized hypoxic activation of cells elicited specific metabolic and gene responses in human microvascular endothelial cells and bone marrow-derived mesenchymal stem cells. A robust demonstration of the compatibility of the device with standard laboratory techniques demonstrates the wide utility of the method. This platform is ideally suited to study real-time cell responses and cell-cell interactions within physiologically relevant oxygen landscapes.

  11. Anti-prion activity generated by a novel vaccine formulation.

    PubMed

    Pilon, John; Loiacono, Christina; Okeson, Danelle; Lund, Sharon; Vercauteren, Kurt; Rhyan, Jack; Miller, Lowell

    2007-12-18

    Chronic wasting disease (CWD) is a transmissible spongiform encephalopathy (TSE) of domestic and wild cervids in North America. To address possible prevention regimens for CWD, we have used a mouse model system and the Rocky Mountain Laboratory (RML) mouse-adapted scrapie prion strain to screen efficacy of potential vaccine candidates. Three peptides derived from the primary amino acid sequence of the prion protein were conjugated to blue carrier protein (BCP) and formulated in an adjuvant containing M. avium subsp. avium. CL57/BL6 mice were vaccinated and boosted with 50 microg of the carrier protein-peptide conjugate formulation; all vaccines produced a humoral immune response as measured by ELISA. Disease challenge with the RML scrapie prion strain revealed anti-prion activity was generated by the vaccine formulations as measured by a delay in clinical disease onset and prolonged survivorship.

  12. CXCL12-mediated murine neural progenitor cell movement requires PI3Kβ activation.

    PubMed

    Holgado, Borja L; Martínez-Muñoz, Laura; Sánchez-Alcañiz, Juan Antonio; Lucas, Pilar; Pérez-García, Vicente; Pérez, Gema; Rodríguez-Frade, José Miguel; Nieto, Marta; Marín, Oscar; Carrasco, Yolanda R; Carrera, Ana C; Alvarez-Dolado, Manuel; Mellado, Mario

    2013-08-01

    The migratory route of neural progenitor/precursor cells (NPC) has a central role in central nervous system development. Although the role of the chemokine CXCL12 in NPC migration has been described, the intracellular signaling cascade involved remains largely unclear. Here we studied the molecular mechanisms that promote murine NPC migration in response to CXCL12, in vitro and ex vivo. Migration was highly dependent on signaling by the CXCL12 receptor, CXCR4. Although the JAK/STAT pathway was activated following CXCL12 stimulation of NPC, JAK activity was not necessary for NPC migration in vitro. Whereas CXCL12 activated the PI3K catalytic subunits p110α and p110β in NPC, only p110β participated in CXCL12-mediated NPC migration. Ex vivo experiments using organotypic slice cultures showed that p110β blockade impaired NPC exit from the medial ganglionic eminence. In vivo experiments using in utero electroporation nonetheless showed that p110β is dispensable for radial migration of pyramidal neurons. We conclude that PI3K p110β is activated in NPC in response to CXCL12, and its activity is necessary for immature interneuron migration to the cerebral cortex.

  13. ActivitySim: large-scale agent based activity generation for infrastructure simulation

    SciTech Connect

    Gali, Emmanuel; Eidenbenz, Stephan; Mniszewski, Sue; Cuellar, Leticia; Teuscher, Christof

    2008-01-01

    The United States' Department of Homeland Security aims to model, simulate, and analyze critical infrastructure and their interdependencies across multiple sectors such as electric power, telecommunications, water distribution, transportation, etc. We introduce ActivitySim, an activity simulator for a population of millions of individual agents each characterized by a set of demographic attributes that is based on US census data. ActivitySim generates daily schedules for each agent that consists of a sequence of activities, such as sleeping, shopping, working etc., each being scheduled at a geographic location, such as businesses or private residences that is appropriate for the activity type and for the personal situation of the agent. ActivitySim has been developed as part of a larger effort to understand the interdependencies among national infrastructure networks and their demand profiles that emerge from the different activities of individuals in baseline scenarios as well as emergency scenarios, such as hurricane evacuations. We present the scalable software engineering principles underlying ActivitySim, the socia-technical modeling paradigms that drive the activity generation, and proof-of-principle results for a scenario in the Twin Cities, MN area of 2.6 M agents.

  14. Simvastatin promotes NPC1-mediated free cholesterol efflux from lysosomes through CYP7A1/LXRα signalling pathway in oxLDL-loaded macrophages.

    PubMed

    Xu, Xiaoyang; Zhang, Aolin; Halquist, Matthew S; Yuan, Xinxu; Henderson, Scott C; Dewey, William L; Li, Pin-Lan; Li, Ningjun; Zhang, Fan

    2017-02-01

    Statins, 3-hydroxyl-3-methylglutaryl coenzyme A reductase inhibitors, are the first-line medications prescribed for the prevention and treatment of coronary artery diseases. The efficacy of statins has been attributed not only to their systemic cholesterol-lowering actions but also to their pleiotropic effects that are unrelated to cholesterol reduction. These pleiotropic effects have been increasingly recognized as essential in statins therapy. This study was designed to investigate the pleiotropic actions of simvastatin, one of the most commonly prescribed statins, on macrophage cholesterol homeostasis with a focus on lysosomal free cholesterol egression. With simultaneous nile red and filipin staining, analysis of confocal/multi-photon imaging demonstrated that simvastatin markedly attenuated unesterified (free) cholesterol buildup in macrophages loaded with oxidized low-density lipoprotein but had little effect in reducing the sizes of cholesteryl ester-containing lipid droplets; the reduction in free cholesterol was mainly attributed to decreases in lysosome-compartmentalized cholesterol. Functionally, the egression of free cholesterol from lysosomes attenuated pro-inflammatory cytokine secretion. It was determined that the reduction of lysosomal free cholesterol buildup by simvastatin was due to the up-regulation of Niemann-Pick C1 (NPC1), a lysosomal residing cholesterol transporter. Moreover, the enhanced enzymatic production of 7-hydroxycholesterol by cytochrome P450 7A1 and the subsequent activation of liver X receptor α underscored the up-regulation of NPC1. These findings reveal a novel pleiotropic effect of simvastatin in affecting lysosomal cholesterol efflux in macrophages and the associated significance in the treatment of atherosclerosis.

  15. An Analysis of Generational Differences Among Active Duty Members

    DTIC Science & Technology

    2004-03-01

    political , and racial lenses. Therefore, it may be difficult to point to an absolute generational group that is defined by a set of shared...group as those born between 1965 and 1978. Finally, the newest generation born after 1979, the Millennials , often given the label Generation Y or Echo...computer technology. Howe and Strauss (2000) define the Millennials as the generation born between 1982 and 2000. The Millennial Cohort, as defined by

  16. Variable phase sine wave generator for active phased arrays

    NASA Astrophysics Data System (ADS)

    Waters, W. M.

    1992-09-01

    A waveform generator is provided for generating a high frequency waveform. A pulse generator provides a pulse train at a low frequency. A pulse converter converts the pulse train into an alternatingly positive and negative groups of pulses. A bandpass filter passes the alternatingly positive and negative groups of pulses in a frequency band centered at the high frequency to output the generated waveform at the high frequency. When the groups of pulses are a pair of pulses, a sine wave is output from the bandpass filter. A pulse delay circuit can be used to variably delay the pulse train and thereby cause a phase change in the generated waveform.

  17. Vitamin E Dietary Supplementation Improves Neurological Symptoms and Decreases c-Abl/p73 Activation in Niemann-Pick C Mice

    PubMed Central

    Marín, Tamara; Contreras, Pablo; Castro, Juan Francisco; Chamorro, David; Balboa, Elisa; Bosch-Morató, Mònica; Muñoz, Francisco J.; Alvarez, Alejandra R.; Zanlungo, Silvana

    2014-01-01

    Niemann-Pick C (NPC) disease is a fatal neurodegenerative disorder characterized by the accumulation of free cholesterol in lysosomes. We have previously reported that oxidative stress is the main upstream stimulus activating the proapoptotic c-Abl/p73 pathway in NPC neurons. We have also observed accumulation of vitamin E in NPC lysosomes, which could lead to a potential decrease of its bioavailability. Our aim was to determine if dietary vitamin E supplementation could improve NPC disease in mice. NPC mice received an alpha-tocopherol (α-TOH) supplemented diet and neurological symptoms, survival, Purkinje cell loss, α-TOH and nitrotyrosine levels, astrogliosis, and the c-Abl/p73 pathway functions were evaluated. In addition, the effect of α-TOH on the c-Abl/p73 pathway was evaluated in an in vitro NPC neuron model. The α-TOH rich diet delayed loss of weight, improved coordination and locomotor function and increased the survival of NPC mice. We found increased Purkinje neurons and α-TOH levels and reduced astrogliosis, nitrotyrosine and phosphorylated p73 in cerebellum. A decrease of c-Abl/p73 activation was also observed in the in vitro NPC neurons treated with α-TOH. In conclusion, our results show that vitamin E can delay neurodegeneration in NPC mice and suggest that its supplementation in the diet could be useful for the treatment of NPC patients. PMID:25079853

  18. Active Control of Fan-Generated Tone Noise

    NASA Technical Reports Server (NTRS)

    Gerhold, Carl H.

    1995-01-01

    This paper reports on an experiment to control the noise radiated from the inlet of a ducted fan using a time domain active adaptive system. The control ,sound source consists of loudspeakers arranged in a ring around the fan duct. The error sensor location is in the fan duct. The purpose of this experiment is to demonstrate that the in-duct error sensor reduces the mode spillover in the far field, thereby increasing the efficiency of the control system. The control system is found to reduce the blade passage frequency tone significantly in the acoustic far field when the mode orders of the noise source and of the control source are the same, when the dominant wave in the duct is a plane wave. The presence of higher order modes in the duct reduces the noise reduction efficiency, particularly near the mode cut-on where the standing wave component is strong, but the control system converges stably. The control system is stable and converges when the first circumferential mode is generated in the duct. The control system is found to reduce the fan noise in the far field on an arc around the fan inlet by as much as 20 dB with none of the sound amplification associated with mode spillover.

  19. On the aerodynamics and performance of active vortex generators

    NASA Technical Reports Server (NTRS)

    Barrett, Ron; Farokhi, Saeed

    1993-01-01

    As a building block in the development of smart lift-enhancement devices, a new concept for flow control using active vortex generators (AVGs) is presented. Ramp, wedge, and doublet wedge (Wheeler) VG configurations are investigated. The AVGs are designed to conform to the surface of the wing section at low alpha. As the section approaches the stall, they are deployed and accordingly, alpha(stall) and C(lmax) are increased. A qualitative analysis of the flow around the various VG configurations was conducted in a low speed wind tunnel at 1.6 ft/s and a Reynolds number of approximately 3400. The results demonstrate that ramp VGs produce vortices that have the longest distance at breakdown. The VGs were also applied to a 25-in. span, 8-in. chord NACA 4415 wing section. Optimization studies were conducted on the spanwise spacing, chordwise position, and size of statically deployed VGs. The test results demonstrate a 14-percent increase in C(lmax) while increasing alpha (stall) by up to 3.

  20. Giant enhancement of second harmonic generation in nonlinear photonic crystals with distributed Bragg reflector mirrors.

    PubMed

    Ren, Ming-Liang; Li, Zhi-Yuan

    2009-08-17

    We theoretically investigate second harmonic generation (SHG) in one-dimensional multilayer nonlinear photonic crystal (NPC) structures with distributed Bragg reflector (DBR) as mirrors. The NPC structures have periodic modulation on both the linear and second-order susceptibility. Three major physical mechanisms, quasi-phase matching (QPM) effect, slow light effect at photonic band gap edges, and cavity effect induced by DBR mirrors can be harnessed to enhance SHG. Selection of appropriate structural parameters can facilitate coexistence of these mechanisms to act collectively and constructively to create very high SHG conversion efficiency with an enhancement by up to seven orders of magnitude compared with the ordinary NPC where only QPM works.

  1. Triptolide inhibits cell growth and GRP78 protein expression but induces cell apoptosis in original and radioresistant NPC cells

    PubMed Central

    Lv, Wuwu; Lai, Chen; Chen, Zhikang; Wang, Ran; Long, Xueying; Feng, Xueping

    2016-01-01

    The radioresistance is the key factor to hamper curative effect and survival of nasopharyngeal carcinoma (NPC) patients. Nature triptolide (TPL) has been found to circumvent drug-resistant effect of cancer, but its effect on NPC radioresistance has been rarely studied. In the present study, the 10 Gy-resistant CNE2 subclones (CNE2-SR) were used as a NPC radioresistant model. The IC50 of TPL in CNE2 and CNE2-SR cells was measured by MTT assay, cell cycle was analyzed by flow cytometry, and protein expression was examined by western blot. Our data showed that TPL treatment decreased the percentage of viable cells, and IC50 value in CNE2 and CNE2-SR cells was 23.6 ± 1.41 nmol/L and 31.2 ± 1.16 nmol/L, respectively. Six Gy was a moderate dosage of X-ray for CNE2, and 25 nM TPL was close to IC50 value of CNE2 and CNE2-SR. Six Gy X-ray and/or 25 nM TPL significantly inhibited tumor growth in nude mice. Furthermore, 6 Gy X-ray and/or 25 nM TPL significantly inhibited cell growth and induced cell apoptosis and M/G2 phase arrest in CNE2 and CNE2-SR cells. Moreover, TPL treatment significantly inhibited the expression of GRP78 protein in CNE2 and CNE2-SR cells. These results suggest that TPL may serve as a potential radiosensitizer agent for NPC treatment. PMID:27391061

  2. Mental Arithmetic Activates Analogic Representations of Internally Generated Sums

    ERIC Educational Resources Information Center

    Kallai, Arava Y.; Schunn, Christian D.; Fiez, Julie A.

    2012-01-01

    The internal representation of numbers generated during calculation has received little attention. Much of the mathematics learning literature focuses on symbolic retrieval of math facts; in contrast, we critically test the hypothesis that internally generated numbers are represented analogically, using an approximate number system. In an fMRI…

  3. Activating Generative Learning in Organizations through Optimizing Relational Strategies

    ERIC Educational Resources Information Center

    Park, Mary Kay

    2010-01-01

    Using a grounded theory method, this dissertation seeks to discover how relationships impact organizational generative learning. An organization is a socially constructed reality and organizational learning is situated in the process of co-participation. To discover the link between relationships and generative learning this study considers the…

  4. Efficacy of Concurrent Chemotherapy for Intermediate Risk NPC in the Intensity-Modulated Radiotherapy Era: a Propensity-Matched Analysis

    PubMed Central

    Zhang, Fan; Zhang, Yuan; Li, Wen-Fei; Liu, Xu; Guo, Rui; Sun, Ying; Lin, Ai-Hua; Chen, Lei; Ma, Jun

    2015-01-01

    This study is to evaluate the efficacy of additional concurrent chemotherapy for intermediate risk (stage II and T3N0M0) NPC patients treated with intensity-modulated radiotherapy (IMRT).440 patients with intermediate risk NPC were studied retrospectively, including 128 patients treated with IMRT alone [radiotherapy group (RT group)] and 312 paitents treated with IMRT plus concurrent chemotherapy [chemoradiotherapy group (CRT group)]. Propensity score matching was carried out to create RT and CRT cohorts equally matched for host and tumor factor. Significantly more severe acute toxicities were observed in the CRT group than in the RT group. Multivariate analyses of 440 patients failed to demonstrate concurrent chemotherapy as an independent prognostic factor for FFS, LR-FFS, and D-FFS. Between the well-matched RT cohort and the CRT cohort, no significant difference was demonstrated in all survival endpoints (FFS: 92.8% versus 91.2%, P = 0.801; LR-FFS: 95.2% versus 94.4%, P = 0.755; D-FFS: 96.4% versus 96.3%, P = 0.803; OS: 98.2% versus 98.9%, P = 0.276). Our results demonstrated that for patients with intermediate risk NPC treated with IMRT, additional concurrent chemotherapy did not provide any significant survival benefit but significantly more severe acute toxicities. However, prospective randomized trials are warranted for the ultimate confirm of our findings. PMID:26611462

  5. Emotion at Work: A Contribution to Third-Generation Cultural-Historical Activity Theory

    ERIC Educational Resources Information Center

    Roth, Wolff-Michael

    2007-01-01

    Second-generation cultural-historical activity theory, which drew its inspiration from Leont'ev's work, constituted an advance over Vygotsky's first-generation theory by explicitly articulating the dialectical relation between individual and collective. As part of an effort to develop third-generation-historical activity theory, I propose in this…

  6. Niemann-Pick Type C2 Protein Mediates Hepatic Stellate Cells Activation by Regulating Free Cholesterol Accumulation.

    PubMed

    Twu, Yuh-Ching; Lee, Tzong-Shyuan; Lin, Yun-Lian; Hsu, Shih-Ming; Wang, Yuan-Hsi; Liao, Chia-Yu; Wang, Chung-Kwe; Liang, Yu-Chih; Liao, Yi-Jen

    2016-07-13

    In chronic liver diseases, regardless of their etiology, the development of fibrosis is the first step toward the progression to cirrhosis, portal hypertension, and hepatocellular carcinoma. Hepatic stellate cells (HSCs) are the main profibrogenic cells that promote the pathogenesis of liver fibrosis, and so it is important to identify the molecules that regulate HSCs activation and liver fibrosis. Niemann-Pick type C2 (NPC2) protein plays an important role in the regulation of intracellular cholesterol homeostasis by directly binding with free cholesterol. However, the roles of NPC2 in HSCs activation and liver fibrosis have not been explored in detail. Since a high-cholesterol diet exacerbates liver fibrosis progression in both rodents and humans, we propose that the expression of NPC2 affects free cholesterol metabolism and regulates HSCs activation. In this study, we found that NPC2 is decreased in both thioacetamide- and carbon tetrachloride-induced liver fibrosis tissues. In addition, NPC2 is expressed in quiescent HSCs, but its activation status is down-regulated. Knockdown of NPC2 in HSC-T6 cells resulted in marked increases in transforming growth factor-β1 (TGF-β1)-induced collagen type 1 α1 (Col1a1), α-smooth muscle actin (α-SMA) expression, and Smad2 phosphorylation. In contrast, NPC2 overexpression decreased TGF-β1-induced HSCs activation. We further demonstrated that NPC2 deficiency significantly increased the accumulation of free cholesterol in HSCs, increasing Col1a1 and α-SMA expression and activating Smad2, and leading to sensitization of HSCs to TGF-β1 activation. In contrast, overexpression of NPC2 decreased U18666A-induced free cholesterol accumulation and inhibited the subsequent HSCs activation. In conclusion, our study has demonstrated that NPC2 plays an important role in HSCs activation by regulating the accumulation of free cholesterol. NPC2 overexpression may thus represent a new treatment strategy for liver fibrosis.

  7. Niemann-Pick Type C2 Protein Mediates Hepatic Stellate Cells Activation by Regulating Free Cholesterol Accumulation

    PubMed Central

    Twu, Yuh-Ching; Lee, Tzong-Shyuan; Lin, Yun-Lian; Hsu, Shih-Ming; Wang, Yuan-Hsi; Liao, Chia-Yu; Wang, Chung-Kwe; Liang, Yu-Chih; Liao, Yi-Jen

    2016-01-01

    In chronic liver diseases, regardless of their etiology, the development of fibrosis is the first step toward the progression to cirrhosis, portal hypertension, and hepatocellular carcinoma. Hepatic stellate cells (HSCs) are the main profibrogenic cells that promote the pathogenesis of liver fibrosis, and so it is important to identify the molecules that regulate HSCs activation and liver fibrosis. Niemann-Pick type C2 (NPC2) protein plays an important role in the regulation of intracellular cholesterol homeostasis by directly binding with free cholesterol. However, the roles of NPC2 in HSCs activation and liver fibrosis have not been explored in detail. Since a high-cholesterol diet exacerbates liver fibrosis progression in both rodents and humans, we propose that the expression of NPC2 affects free cholesterol metabolism and regulates HSCs activation. In this study, we found that NPC2 is decreased in both thioacetamide- and carbon tetrachloride-induced liver fibrosis tissues. In addition, NPC2 is expressed in quiescent HSCs, but its activation status is down-regulated. Knockdown of NPC2 in HSC-T6 cells resulted in marked increases in transforming growth factor-β1 (TGF-β1)-induced collagen type 1 α1 (Col1a1), α-smooth muscle actin (α-SMA) expression, and Smad2 phosphorylation. In contrast, NPC2 overexpression decreased TGF-β1-induced HSCs activation. We further demonstrated that NPC2 deficiency significantly increased the accumulation of free cholesterol in HSCs, increasing Col1a1 and α-SMA expression and activating Smad2, and leading to sensitization of HSCs to TGF-β1 activation. In contrast, overexpression of NPC2 decreased U18666A-induced free cholesterol accumulation and inhibited the subsequent HSCs activation. In conclusion, our study has demonstrated that NPC2 plays an important role in HSCs activation by regulating the accumulation of free cholesterol. NPC2 overexpression may thus represent a new treatment strategy for liver fibrosis. PMID:27420058

  8. Genetic predisposition factors and nasopharyngeal carcinoma risk: a review of epidemiological association studies, 2000-2011: Rosetta Stone for NPC: genetics, viral infection, and other environmental factors.

    PubMed

    Hildesheim, Allan; Wang, Cheng-Ping

    2012-04-01

    While infection with Epstein-Barr virus (EBV) is known to be an essential risk factor for the development of nasopharyngeal carcinoma (NPC), other co-factors including genetic factors are thought to play an important role. In this review, we summarize association studies conducted over the past decade to evaluate the role of genetic polymorphisms in NPC development. A review of the literature identified close to 100 studies, including 3 genome-wide association studies (GWAS), since 2000 that evaluated genetic polymorphisms and NPC risk in at least 100 NPC cases and 100 controls. Consistent evidence for associations were reported for a handful of genes, including immune-related HLA Class I genes, DNA repair gene RAD51L1, cell cycle control genes MDM2 and TP53, and cell adhesion/migration gene MMP2. However, for most of the genes evaluated, there was no effort to replicate findings and studies were largely modest in size, typically consisting of no more than a few hundred cases and controls. The small size of most studies, and the lack of attempts at replication have limited progress in understanding the genetics of NPC. Moving forward, if we are to advance our understanding of genetic factors involved in the development of NPC, and of the impact of gene-gene and gene-environment interations in the development of this disease, consortial efforts that pool across multiple, well-designed and coordinated efforts will most likely be required.

  9. Cortical hyperpolarization-activated depolarizing current takes part in the generation of focal paroxysmal activities

    PubMed Central

    Timofeev, Igor; Bazhenov, Maxim; Sejnowski, Terrence; Steriade, Mircea

    2002-01-01

    During paroxysmal neocortical oscillations, sudden depolarization leading to the next cycle occurs when the majority of cortical neurons are hyperpolarized. Both the Ca2+-dependent K+ currents (IK(Ca)) and disfacilitation play critical roles in the generation of hyperpolarizing potentials. In vivo experiments and computational models are used here to investigate whether the hyperpolarization-activated depolarizing current (Ih) in cortical neurons also contributes to the generation of paroxysmal onsets. Hyperpolarizing current pulses revealed a depolarizing sag in ≈20% of cortical neurons. Intracellular recordings from glial cells indirectly indicated an increase in extracellular potassium concentration ([K+]o) during paroxysmal activities, leading to a positive shift in the reversal potential of K+-mediated currents, including Ih. In the paroxysmal neocortex, ≈20% of neurons show repolarizing potentials originating from hyperpolarizations associated with depth-electroencephalogram positive waves of spike-wave complexes. The onset of these repolarizing potentials corresponds to maximal [K+]o as estimated from dual simultaneous impalements from neurons and glial cells. Computational models showed how, after the increased [K+]o, the interplay between Ih, IK(Ca), and a persistent Na+ current, INa(P), could organize paroxysmal oscillations at a frequency of 2–3 Hz. PMID:12089324

  10. New murine Niemann-Pick type C models bearing a pseudoexon-generating mutation recapitulate the main neurobehavioural and molecular features of the disease.

    PubMed

    Gómez-Grau, Marta; Albaigès, Júlia; Casas, Josefina; Auladell, Carme; Dierssen, Mara; Vilageliu, Lluïsa; Grinberg, Daniel

    2017-02-07

    Niemann-Pick disease type C (NPC) is a rare neurovisceral disease caused mainly by mutations in the NPC1 gene. This autosomal recessive lysosomal disorder is characterised by the defective lysosomal secretion of cholesterol and sphingolipids. No effective therapy exists for the disease. We previously described a deep intronic point mutation (c.1554-1009 G > A) in NPC1 that generated a pseudoexon, which could be corrected at the cellular level using antisense oligonucleotides. Here, we describe the generation of two mouse models bearing this mutation, one in homozygosity and the other in compound heterozygosity with the c.1920delG mutation. Both the homozygotes for the c.1554-1009 G > A mutation and the compound heterozygotes recapitulated the hallmarks of NPC. Lipid analysis revealed accumulation of cholesterol in the liver and sphingolipids in the brain, with both types of transgenic mice displaying tremor and ataxia at 7-8 weeks of age. Behavioural tests showed motor impairment, hyperactivity, reduced anxiety-like behaviour and impaired learning and memory performances, features consistent with those reported previously in NPC animal models and human patients. These mutant mice, the first NPC models bearing a pseudoexon-generating mutation, could be suitable for assessing the efficacy of specific splicing-targeted therapeutic strategies against NPC.

  11. New murine Niemann-Pick type C models bearing a pseudoexon-generating mutation recapitulate the main neurobehavioural and molecular features of the disease

    PubMed Central

    Gómez-Grau, Marta; Albaigès, Júlia; Casas, Josefina; Auladell, Carme; Dierssen, Mara; Vilageliu, Lluïsa; Grinberg, Daniel

    2017-01-01

    Niemann-Pick disease type C (NPC) is a rare neurovisceral disease caused mainly by mutations in the NPC1 gene. This autosomal recessive lysosomal disorder is characterised by the defective lysosomal secretion of cholesterol and sphingolipids. No effective therapy exists for the disease. We previously described a deep intronic point mutation (c.1554-1009 G > A) in NPC1 that generated a pseudoexon, which could be corrected at the cellular level using antisense oligonucleotides. Here, we describe the generation of two mouse models bearing this mutation, one in homozygosity and the other in compound heterozygosity with the c.1920delG mutation. Both the homozygotes for the c.1554-1009 G > A mutation and the compound heterozygotes recapitulated the hallmarks of NPC. Lipid analysis revealed accumulation of cholesterol in the liver and sphingolipids in the brain, with both types of transgenic mice displaying tremor and ataxia at 7–8 weeks of age. Behavioural tests showed motor impairment, hyperactivity, reduced anxiety-like behaviour and impaired learning and memory performances, features consistent with those reported previously in NPC animal models and human patients. These mutant mice, the first NPC models bearing a pseudoexon-generating mutation, could be suitable for assessing the efficacy of specific splicing-targeted therapeutic strategies against NPC. PMID:28167839

  12. Does changing from a first generation antipsychotic (perphenazin) to a second generation antipsychotic (risperidone) alter brain activation and motor activity? A case report

    PubMed Central

    2013-01-01

    Background In patients with schizophrenia, altered brain activation and motor activity levels are central features, reflecting cognitive impairments and negative symptoms, respectively. Newer studies using nonlinear methods have addressed the severe disturbances in neurocognitive functioning that is regarded as one of the core features of schizophrenia. Our aim was to compare brain activation and motor activity in a patient during pharmacological treatment that was switched from a first- to a second-generation antipsychotic drug. We hypothesised that this change of medication would increase level of responding in both measures. Case presentation We present the case of a 53-year-old male with onset of severe mental illness in adolescence, ICD-10 diagnosed as schizophrenia of paranoid type, chronic form. We compared brain activation and motor activity in this patient during pharmacological treatment with a first-generation (perphenazin), and later switched to a second-generation (risperidone) antipsychotic drug. We used functional magnetic resonance imaging (fMRI) to measure brain activation and wrist worn actigraphy to measure motor activity. Conclusion Our study showed that brain activation decreased in areas critical for cognitive functioning in this patient, when changing from a first to a second generation antipsychotic drug. However the mean motor activity level was unchanged, although risperidone reduced variability, particularly short-term variability from minute to minute. Compared to the results from previous studies, the present findings indicate that changing to a second-generation antipsychotic alters variability measures towards that seen in a control group, but with reduced brain activation, which was an unexpected finding. PMID:23648137

  13. Generations.

    PubMed

    Chambers, David W

    2005-01-01

    Groups naturally promote their strengths and prefer values and rules that give them an identity and an advantage. This shows up as generational tensions across cohorts who share common experiences, including common elders. Dramatic cultural events in America since 1925 can help create an understanding of the differing value structures of the Silents, the Boomers, Gen Xers, and the Millennials. Differences in how these generations see motivation and values, fundamental reality, relations with others, and work are presented, as are some applications of these differences to the dental profession.

  14. CRISPR/Cas9-Mediated Generation of Niemann-Pick C1 Knockout Cell Line.

    PubMed

    Du, Ximing; Lukmantara, Ivan; Yang, Hongyuan

    2017-01-01

    Generating a cholesterol storage phenotype of Niemann-Pick Type C (NPC) disease is important for investigating the mechanisms of intracellular cholesterol trafficking, as well as screening drugs for potential treatment of NPC disease. The use of the CRISPR/Cas9 technology to knockout specific genes within the genome of mammals has become routine in the past few years. Here, we describe a protocol for producing a cellular NPC cholesterol storage phenotype in HeLa cells using the CRISPR-Cas9 system to disrupt the NPC1 gene. The protocol details the steps for single guide RNA oligo cloning, cell colony selection, and cell line verification by filipin staining and immunoblotting.

  15. Biological activity of photoproducts of merocyanine 540 generated by laser-light activation

    NASA Astrophysics Data System (ADS)

    Gulliya, Kirpal S.; Chanh, Tran C.; Pervaiz, Shazib; Harriman, Anthony; Matthews, James Lester

    1992-08-01

    Controlled exposure of photoactive compounds to light prior to their use in biological targets results in the formation of heretofore unknown photoproducts. This process of photoproduct generation, termed "preactivation," renders the photactive compound capable of systemic use without further dependence on light. Preactivation of mercyanin 540 (MC540) and several other photoactive compounds is achievable by exposure to CW and pulse laser radiation. The singlet oxygen generated at excited states attacks the dye molucule itself, resulting in the formation of biologically active photoproducts. For preactivated MC540 (photoproducts of MC540) generated by exposure to argon laser light (514 nm) and light from free-electron laser, we have demonstrated its effectiveness in selective killing of certain types of cultured tumor cells as well as human immunodeficiency virus type 1 (HIV-1) with very low, if any, damage to normal cells and tisues. For example, approximately 90% of the Burkitt's lymphoma Daudi cells and HL-60 leukemic cells are killed by preactivated MC540 at a concentration of 120 μg/ml. A two-hour treatment of cultured cells with buthionine sulfoxamine followed by the treatement with preactivated MC540 reults in 99.99% inhibition of clonogenic tumor stem cell growth. We also have demonstrated that preactivated MC540 is very effective in killing cell-free and cell-associated HIV-1. It also is very effective in killing HIV-1 and simian immunodeficiency virus (SIV) in virus-infected blood in vitro as determined by reverse transcriptase, P24, P17, core antigen expression and synctium formation. Treatment of HIV-1 with preactivated MC540 renders the treated HIV-1 incapable of binding to CD4 target molecules on T cells as determined by immunofluorescence and radioimmunoprecipitation assays. In vivo toxicology studies show that preactivated MC540 is very well tolerated and does not produce any signs of adverse reaction at the therapeutic doses, as determined by

  16. Sensing network for electromagnetic fields generated by seismic activities

    NASA Astrophysics Data System (ADS)

    Gershenzon, Naum I.; Bambakidis, Gust; Ternovskiy, Igor V.

    2014-06-01

    The sensors network is becoming prolific and play now increasingly more important role in acquiring and processing information. Cyber-Physical Systems are focusing on investigation of integrated systems that includes sensing, networking, and computations. The physics of the seismic measurement and electromagnetic field measurement requires special consideration how to design electromagnetic field measurement networks for both research and detection earthquakes and explosions along with the seismic measurement networks. In addition, the electromagnetic sensor network itself could be designed and deployed, as a research tool with great deal of flexibility, the placement of the measuring nodes must be design based on systematic analysis of the seismic-electromagnetic interaction. In this article, we review the observations of the co-seismic electromagnetic field generated by earthquakes and man-made sources such as vibrations and explosions. The theoretical investigation allows the distribution of sensor nodes to be optimized and could be used to support existing geological networks. The placement of sensor nodes have to be determined based on physics of electromagnetic field distribution above the ground level. The results of theoretical investigations of seismo-electromagnetic phenomena are considered in Section I. First, we compare the relative contribution of various types of mechano-electromagnetic mechanisms and then analyze in detail the calculation of electromagnetic fields generated by piezomagnetic and electrokinetic effects.

  17. Educating for Political Activity: A Younger Generational Response

    ERIC Educational Resources Information Center

    Mac an Ghaill, Mairtin

    2010-01-01

    This paper is a response to Professor Chitty's "Educational Review" Guest Lecture article, "Educating for political activity". I address the three sections of his paper: a global and national-based politics of war, corporate manipulation and parliamentary scandals. This provides a basis to draw upon empirical material from a…

  18. Morphological Transformation and Force Generation of Active Cytoskeletal Networks

    PubMed Central

    Maruri, Daniel; Kamm, Roger D.

    2017-01-01

    Cells assemble numerous types of actomyosin bundles that generate contractile forces for biological processes, such as cytokinesis and cell migration. One example of contractile bundles is a transverse arc that forms via actomyosin-driven condensation of actin filaments in the lamellipodia of migrating cells and exerts significant forces on the surrounding environments. Structural reorganization of a network into a bundle facilitated by actomyosin contractility is a physiologically relevant and biophysically interesting process. Nevertheless, it remains elusive how actin filaments are reoriented, buckled, and bundled as well as undergo tension buildup during the structural reorganization. In this study, using an agent-based computational model, we demonstrated how the interplay between the density of myosin motors and cross-linking proteins and the rigidity, initial orientation, and turnover of actin filaments regulates the morphological transformation of a cross-linked actomyosin network into a bundle and the buildup of tension occurring during the transformation. PMID:28114384

  19. Scientific and legal perspectives on science generated for regulatory activities.

    PubMed

    Henry, Carol J; Conrad, James W

    2008-01-01

    This article originated from a conference that asked "Should scientific work conducted for purposes of advocacy before regulatory agencies or courts be judged by the same standards as science conducted for other purposes?" In the article, which focuses on the regulatory advocacy context, we argue that it can be and should be. First, we describe a set of standards and practices currently being used to judge the quality of scientific research and testing and explain how these standards and practices assist in judging the quality of research and testing regardless of why the work was conducted. These standards and practices include the federal Information Quality Act, federal Good Laboratory Practice standards, peer review, disclosure of funding sources, and transparency in research policies. The more that scientific information meets these standards and practices, the more likely it is to be of high quality, reliable, reproducible, and credible. We then explore legal issues that may be implicated in any effort to create special rules for science conducted specifically for a regulatory proceeding. Federal administrative law does not provide a basis for treating information in a given proceeding differently depending on its source or the reason for which it was generated. To the contrary, this law positively assures that interested persons have the right to offer their technical expertise toward the solution of regulatory problems. Any proposal to subject scientific information generated for the purpose of a regulatory proceeding to more demanding standards than other scientific information considered in that proceeding would clash with this law and would face significant administrative complexities. In a closely related example, the U.S. Environmental Protection Agency considered but abandoned a program to implement standards aimed at "external" information.

  20. Probabilistic Analysis of Activation Volumes Generated During Deep Brain Stimulation

    PubMed Central

    Butson, Christopher R.; Cooper, Scott E.; Henderson, Jaimie M.; Wolgamuth, Barbara; McIntyre, Cameron C.

    2010-01-01

    Deep brain stimulation (DBS) is an established therapy for the treatment of Parkinson’s disease (PD) and shows great promise for the treatment of several other disorders. However, while the clinical analysis of DBS has received great attention, a relative paucity of quantitative techniques exists to define the optimal surgical target and most effective stimulation protocol for a given disorder. In this study we describe a methodology that represents an evolutionary addition to the concept of a probabilistic brain atlas, which we call a probabilistic stimulation atlas (PSA). We outline steps to combine quantitative clinical outcome measures with advanced computational models of DBS to identify regions where stimulation-induced activation could provide the best therapeutic improvement on a per-symptom basis. While this methodology is relevant to any form of DBS, we present example results from subthalamic nucleus (STN) DBS for PD. We constructed patient-specific computer models of the volume of tissue activated (VTA) for 163 different stimulation parameter settings which were tested in six patients. We then assigned clinical outcome scores to each VTA and compiled all of the VTAs into a PSA to identify stimulation-induced activation targets that maximized therapeutic response with minimal side effects. The results suggest that selection of both electrode placement and clinical stimulation parameter settings could be tailored to the patient’s primary symptoms using patient-specific models and PSAs. PMID:20974269

  1. Probabilistic analysis of activation volumes generated during deep brain stimulation.

    PubMed

    Butson, Christopher R; Cooper, Scott E; Henderson, Jaimie M; Wolgamuth, Barbara; McIntyre, Cameron C

    2011-02-01

    Deep brain stimulation (DBS) is an established therapy for the treatment of Parkinson's disease (PD) and shows great promise for the treatment of several other disorders. However, while the clinical analysis of DBS has received great attention, a relative paucity of quantitative techniques exists to define the optimal surgical target and most effective stimulation protocol for a given disorder. In this study we describe a methodology that represents an evolutionary addition to the concept of a probabilistic brain atlas, which we call a probabilistic stimulation atlas (PSA). We outline steps to combine quantitative clinical outcome measures with advanced computational models of DBS to identify regions where stimulation-induced activation could provide the best therapeutic improvement on a per-symptom basis. While this methodology is relevant to any form of DBS, we present example results from subthalamic nucleus (STN) DBS for PD. We constructed patient-specific computer models of the volume of tissue activated (VTA) for 163 different stimulation parameter settings which were tested in six patients. We then assigned clinical outcome scores to each VTA and compiled all of the VTAs into a PSA to identify stimulation-induced activation targets that maximized therapeutic response with minimal side effects. The results suggest that selection of both electrode placement and clinical stimulation parameter settings could be tailored to the patient's primary symptoms using patient-specific models and PSAs.

  2. Phospholipid-esterified eicosanoids are generated in agonist-activated human platelets and enhance tissue factor-dependent thrombin generation.

    PubMed

    Thomas, Christopher P; Morgan, Lloyd T; Maskrey, Benjamin H; Murphy, Robert C; Kühn, Hartmut; Hazen, Stanley L; Goodall, Alison H; Hamali, Hassan A; Collins, Peter W; O'Donnell, Valerie B

    2010-03-05

    Here, a group of specific lipids, comprising phosphatidylethanolamine (PE)- or phosphatidylcholine (PC)-esterified 12S-hydroxyeicosatetraenoic acid (12S-HETE), generated by 12-lipoxygenase was identified and characterized. 12S-HETE-PE/PCs were formed within 5 min of activation by thrombin, ionophore, or collagen. Esterified HETE levels generated in response to thrombin were 5.85 +/- 1.42 (PE) or 18.35 +/- 4.61 (PC), whereas free was 65.5 +/- 17.6 ng/4 x 10(7) cells (n = 5 separate donors, mean +/- S.E.). Their generation was stimulated by triggering protease-activated receptors-1 and -4 and signaling via Ca(2+) mobilization secretory phospholipase A2, platelet-activating factor-acetylhydrolase, src tyrosine kinases, and protein kinase C. Stable isotope labeling showed that they form predominantly by esterification that occurs on the same time scale as free acid generation. Unlike free 12S-HETE that is secreted, esterified HETEs remain cell-associated, with HETE-PEs migrating to the outside of the plasma membrane. 12-Lipoxygenase inhibition attenuated externalization of native PE and phosphatidylserine and HETE-PEs. Platelets from a patient with the bleeding disorder, Scott syndrome, did not externalize HETE-PEs, and liposomes supplemented with HETE-PC dose-dependently enhanced tissue factor-dependent thrombin generation in vitro. This suggests a role for these novel lipids in promoting coagulation. Thus, oxidized phospholipids form by receptor/agonist mechanisms, not merely as an undesirable consequence of vascular and inflammatory disease.

  3. Impact-generated Hydrothermal Activity at the Chicxulub Crater

    NASA Astrophysics Data System (ADS)

    Kring, D. A.; Zurcher, L.; Abramov, O.

    2007-05-01

    Borehole samples recovered from PEMEX exploration boreholes and an ICDP scientific borehole indicate the Chicxulub impact event generated hydrothermal alteration throughout a large volume of the Maya Block beneath the crater floor and extending across the bulk of the ~180 km diameter crater. The first indications of hydrothermal alteration were observed in the crater discovery samples from the Yucatan-6 borehole and manifest itself in the form of anhydrite and quartz veins. Continuous core from the Yaxcopoil-1 borehole reveal a more complex and temporally extensive alteration sequence: following a brief period at high temperatures, impact- melt-bearing polymict breccias and a thin, underlying unit of impact melt were subjected to metasomatism, producing alkali feldspar, sphene, apatite, and magnetite. As the system continued to cool, smectite-series phyllosilicates appeared. A saline solution was involved. Stable isotopes suggest the fluid was dominated by a basinal brine created mostly from existing groundwater of the Yucatan Peninsula, although contributions from down-welling water also occurred in some parts of the system. Numerical modeling of the hydrothermal system suggests circulation occurred for 1.5 to 2.3 Myr, depending on the permeability of the system. Our understanding of the hydrothermal system, however, is still crude. Additional core recovery projects, particularly into the central melt sheet, are needed to better evaluate the extent and duration of hydrothermal alteration.

  4. Deterministic generation of remote entanglement with active quantum feedback

    SciTech Connect

    Martin, Leigh; Motzoi, Felix; Li, Hanhan; Sarovar, Mohan; Whaley, K. Birgitta

    2015-12-10

    We develop and study protocols for deterministic remote entanglement generation using quantum feedback, without relying on an entangling Hamiltonian. In order to formulate the most effective experimentally feasible protocol, we introduce the notion of average-sense locally optimal feedback protocols, which do not require real-time quantum state estimation, a difficult component of real-time quantum feedback control. We use this notion of optimality to construct two protocols that can deterministically create maximal entanglement: a semiclassical feedback protocol for low-efficiency measurements and a quantum feedback protocol for high-efficiency measurements. The latter reduces to direct feedback in the continuous-time limit, whose dynamics can be modeled by a Wiseman-Milburn feedback master equation, which yields an analytic solution in the limit of unit measurement efficiency. Our formalism can smoothly interpolate between continuous-time and discrete-time descriptions of feedback dynamics and we exploit this feature to derive a superior hybrid protocol for arbitrary nonunit measurement efficiency that switches between quantum and semiclassical protocols. Lastly, we show using simulations incorporating experimental imperfections that deterministic entanglement of remote superconducting qubits may be achieved with current technology using the continuous-time feedback protocol alone.

  5. Deterministic generation of remote entanglement with active quantum feedback

    DOE PAGES

    Martin, Leigh; Motzoi, Felix; Li, Hanhan; ...

    2015-12-10

    We develop and study protocols for deterministic remote entanglement generation using quantum feedback, without relying on an entangling Hamiltonian. In order to formulate the most effective experimentally feasible protocol, we introduce the notion of average-sense locally optimal feedback protocols, which do not require real-time quantum state estimation, a difficult component of real-time quantum feedback control. We use this notion of optimality to construct two protocols that can deterministically create maximal entanglement: a semiclassical feedback protocol for low-efficiency measurements and a quantum feedback protocol for high-efficiency measurements. The latter reduces to direct feedback in the continuous-time limit, whose dynamics can bemore » modeled by a Wiseman-Milburn feedback master equation, which yields an analytic solution in the limit of unit measurement efficiency. Our formalism can smoothly interpolate between continuous-time and discrete-time descriptions of feedback dynamics and we exploit this feature to derive a superior hybrid protocol for arbitrary nonunit measurement efficiency that switches between quantum and semiclassical protocols. Lastly, we show using simulations incorporating experimental imperfections that deterministic entanglement of remote superconducting qubits may be achieved with current technology using the continuous-time feedback protocol alone.« less

  6. Multiple rare variants in NPC1L1 associated with reduced sterol absorption and plasma low-density lipoprotein levels

    PubMed Central

    Cohen, Jonathan C.; Pertsemlidis, Alexander; Fahmi, Saleemah; Esmail, Sophie; Vega, Gloria L.; Grundy, Scott M.; Hobbs, Helen H.

    2006-01-01

    An approach to understand quantitative traits was recently proposed based on the finding that nonsynonymous (NS) sequence variants in certain genes are preferentially enriched at one extreme of the population distribution. The NS variants, although individually rare, are cumulatively frequent and influence quantitative traits, such as plasma lipoprotein levels. Here, we use the NS variant technique to demonstrate that genetic variation in NPC1L1 contributes to variability in cholesterol absorption and plasma levels of low-density lipoproteins (LDLs). The ratio of plasma campesterol (a plant sterol) to lathosterol (a cholesterol precursor) was used to estimate relative cholesterol absorption in a population-based study. Nonsynonymous sequence variations in NPC1L1 were five times more common in low absorbers (n = 26 of 256) than in high absorbers (n = 5 of 256) (P < 0.001). The rare variants identified in low absorbers were found in 6% of 1,832 African-Americans and were associated with lower plasma levels of LDL cholesterol (LDL-C) (96 ± 36 mg/dl vs. 105 ± 36 mg/dl; P = 0.005). These data, together with prior findings, reveal a genetic architecture for LDL-C levels that does not conform to current models for quantitative traits and indicate that a significant fraction of genetic variance in LDL-C is due to multiple alleles with modest effects that are present at low frequencies in the population. PMID:16449388

  7. Compact and highly active next-generation libraries for CRISPR-mediated gene repression and activation

    PubMed Central

    Horlbeck, Max A; Gilbert, Luke A; Villalta, Jacqueline E; Adamson, Britt; Pak, Ryan A; Chen, Yuwen; Fields, Alexander P; Park, Chong Yon; Corn, Jacob E; Kampmann, Martin; Weissman, Jonathan S

    2016-01-01

    We recently found that nucleosomes directly block access of CRISPR/Cas9 to DNA (Horlbeck et al., 2016). Here, we build on this observation with a comprehensive algorithm that incorporates chromatin, position, and sequence features to accurately predict highly effective single guide RNAs (sgRNAs) for targeting nuclease-dead Cas9-mediated transcriptional repression (CRISPRi) and activation (CRISPRa). We use this algorithm to design next-generation genome-scale CRISPRi and CRISPRa libraries targeting human and mouse genomes. A CRISPRi screen for essential genes in K562 cells demonstrates that the large majority of sgRNAs are highly active. We also find CRISPRi does not exhibit any detectable non-specific toxicity recently observed with CRISPR nuclease approaches. Precision-recall analysis shows that we detect over 90% of essential genes with minimal false positives using a compact 5 sgRNA/gene library. Our results establish CRISPRi and CRISPRa as premier tools for loss- or gain-of-function studies and provide a general strategy for identifying Cas9 target sites. DOI: http://dx.doi.org/10.7554/eLife.19760.001 PMID:27661255

  8. Generation and remote delivery of plasma activated species

    NASA Astrophysics Data System (ADS)

    Maguire, Paul; Mahony, Charles; Kelsey, Colin; Rutherford, David; Mariotti, Davide; Macias-Montero, Manuel; Perez-Martin, Fatima; Diver, Declan

    2016-09-01

    Plasma interactions with microdroplets offer new opportunities to deliver active chemical agents and nanoparticles to remote substrates downstream with many potential applications from cancer theranostics and wound healing in biomedicine, gentle food decontamination and seed germination in plasma agriculture to catalyst production and photonic structures fabrication, among others. We demonstrate plasma-liquid based pristine nanomaterials synthesis in flight and subsequent delivery up to 120mm from the atmospheric pressure plasma source. Monosized and non-aggregating metal nanoparticles are formed in the rf plasma in less than 100us, representing an increase in precursor reduction rate that is many (>4) orders of magnitude faster than that observed with standard colloidal chemistry or via high energy radiolytic techniques. Also the collection and purification limitations of the latter are avoided. Plasma activated liquid including OH radicals and H2O2 are transported over 120mm and have demonstrated high efficacy bacterial decontamination. These results will be compared with charge species and radical transport from the rf plasma without microdroplets. Reaction models based on high solvated surface electron concentrations will be presented. Funding from EPSRC acknowledged (Grants EP/K006088/1 and EP/K006142/1).

  9. Reversible activation of the neutrophil superoxide generating system by hexachlorocyclohexane: correlation with effects on a subcellular superoxide-generating fraction.

    PubMed

    English, D; Schell, M; Siakotos, A; Gabig, T G

    1986-07-01

    gamma-Hexachlorocyclohexane was found to exert profound effects on the phosphatidylinositol cycle, cytosolic calcium level, and the respiratory burst of human neutrophils. Exposure of neutrophils prelabelled with 32P to 4 X 10(-4) M gamma-hexachlorocyclohexane almost tripled radioactivity in phosphatidic acid and correspondingly decreased radioactivity in phosphatidylinositol 4,5 bisphosphate. Under similar conditions, gamma-hexachlorocyclohexane evoked the generation of superoxide at a rate of over 11 nmol/min/10(6) cells and more than doubled cytosolic-free calcium concentration as monitored by Quin-2 fluorescence. Because intermediates of the phosphatidylinositol cycle, via increases in available calcium levels or activated protein kinase C, are considered potential second messengers for activation of the NADPH-dependent O-2-generating system, we compared neutrophil responses to gamma-hexachlorocyclohexane with responses to phorbol myristate acetate, an activator of protein kinase C with well known effects on neutrophils. Like phorbol myristate acetate, gamma-hexachlorocyclohexane induced neutrophil degranulation but was not an effective chemotactic stimulus. The ability of gamma-hexachlorocyclohexane to induce a pattern of oxidative activation in neutrophil cytoplasts similar to that in intact cells indicated that concurrent degranulation was not required for sustained O-2 generation in response to this agent. When neutrophils or neutrophil cytoplasts exposed to gamma-hexachlorocyclohexane were centrifuged and resuspended in stimulus-free medium, O-2 generation ceased entirely but could be reinitiated by addition of the same stimulus. This finding was in contrast to the continued O-2 production by phorbol myristate acetate-stimulated neutrophils similarly washed and resuspended in stimulus-free medium. Unlike subcellular fractions of phorbol myristate acetate-stimulated neutrophils, corresponding fractions prepared from gamma

  10. Active fiber composites for the generation of Lamb waves.

    PubMed

    Birchmeier, M; Gsell, D; Juon, M; Brunner, A J; Paradies, R; Dual, J

    2009-01-01

    Active fiber composites (AFC) are thin and conformable transducer elements with orthotropic material properties, since they are made of one layer of piezoelectric ceramic fibers. They are suitable for applications in structural health monitoring systems (SHM) with acoustic non-destructive testing methods (NDT). In the presented work the transfer behavior of an AFC as an emitter of transient elastic waves in plate-like structures is investigated. The wave field emitted by an AFC surface bonded on an isotropic plate was simulated with the finite-difference method. The model includes the piezoelectric element and the plate and allows the simulation of the elastic wave propagation. For comparison with the model experiments using a laser interferometer for non-contact measurements of particle velocities at different points around the AFC on the surface of the plate were performed. Transfer functions defined as the ratio of the electric voltage excitation signal and the resulting surface velocity at a specific point are separately determined for the two fundamental Lamb wave modes. In order to take the orthotropic behavior of the AFC into account the transfer functions are determined for several points around the AFC. Results show that the AFC is capable to excite the fundamental symmetric and antisymmetric Lamb wave mode. The antisymmetric mode is mainly radiated in the direction of the piezoelectric fibers, while the symmetric mode is spread over a larger angle. The amplitudes of the emitted waves depend on the frequency of the excitation as well as on the geometric dimensions of the transducer.

  11. Increased Activity and Altered Subcellular Distribution of Lysosomal Enzymes Determine Neuronal Vulnerability in Niemann-Pick Type C1-Deficient Mice

    PubMed Central

    Amritraj, Asha; Peake, Kyle; Kodam, Anitha; Salio, Chiara; Merighi, Adalberto; Vance, Jean E.; Kar, Satyabrata

    2009-01-01

    Niemann-Pick disease type C (NPC), caused by mutations in the Npc1 or Npc2 genes, is a progressive neurodegenerative disorder characterized by intracellular accumulation/redistribution of cholesterol in a number of tissues including the brain. This is accompanied by a severe loss of neurons in selected brain regions. In this study, we evaluated the role of lysosomal enzymes, cathepsins B and D, in determining neuronal vulnerability in NPC1-deficient (Npc1−/−) mouse brains. Our results showed that Npc1−/− mice exhibit an age-dependent degeneration of neurons in the cerebellum but not in the hippocampus. The cellular level/expression and activity of cathepsins B and D are increased more predominantly in the cerebellum than in the hippocampus of Npc1−/− mice. In addition, the cytosolic levels of cathepsins, cytochrome c, and Bax2 are higher in the cerebellum than in the hippocampus of Npc1−/− mice, suggesting a role for these enzymes in the degeneration of neurons. This suggestion is supported by our observation that degeneration of cultured cortical neurons treated with U18666A, which induces an NPC1-like phenotype at the cellular level, can be attenuated by inhibition of cathepsin B or D enzyme activity. These results suggest that the increased level/activity and altered subcellular distribution of cathepsins may be associated with the underlying cause of neuronal vulnerability in Npc1−/− brains. Therefore, their inhibitors may have therapeutic potential in attenuating NPC pathology. PMID:19893049

  12. APP overexpression in the absence of NPC1 exacerbates metabolism of amyloidogenic proteins of Alzheimer's disease

    PubMed Central

    Maulik, Mahua; Peake, Kyle; Chung, JiYun; Wang, Yanlin; Vance, Jean E.; Kar, Satyabrata

    2015-01-01

    Amyloid-β (Aβ) peptides originating from β-amyloid precursor protein (APP) are critical in Alzheimer's disease (AD). Cellular cholesterol levels/distribution can regulate production and clearance of Aβ peptides, albeit with contradictory outcomes. To better understand the relationship between cholesterol homeostasis and APP/Aβ metabolism, we have recently generated a bigenic ANPC mouse line overexpressing mutant human APP in the absence of Niemann-Pick type C-1 protein required for intracellular cholesterol transport. Using this unique bigenic ANPC mice and complementary stable N2a cells, we have examined the functional consequences of cellular cholesterol sequestration in the endosomal–lysosomal system, a major site of Aβ production, on APP/Aβ metabolism and its relation to neuronal viability. Levels of APP C-terminal fragments (α-CTF/β-CTF) and Aβ peptides, but not APP mRNA/protein or soluble APPα/APPβ, were increased in ANPC mouse brains and N2a-ANPC cells. These changes were accompanied by reduced clearance of peptides and an increased level/activity of γ-secretase, suggesting that accumulation of APP-CTFs is due to decreased turnover, whereas increased Aβ levels may result from a combination of increased production and decreased turnover. APP-CTFs and Aβ peptides were localized primarily in early-/late-endosomes and to some extent in lysosomes/autophagosomes. Cholesterol sequestration impaired endocytic-autophagic-lysosomal, but not proteasomal, clearance of APP-CTFs/Aβ peptides. Moreover, markers of oxidative stress were increased in vulnerable brain regions of ANPC mice and enhanced β-CTF/Aβ levels increased susceptibility of N2a-ANPC cells to H2O2-induced toxicity. Collectively, our results show that cellular cholesterol sequestration plays a key role in APP/Aβ metabolism and increasing neuronal vulnerability to oxidative stress in AD-related pathology. PMID:26433932

  13. Leveling the Playing Field: First Generation Korean American Males and School Based Extracurricular Activities

    ERIC Educational Resources Information Center

    Levy, Corey

    2010-01-01

    This study examined the manner in which extracurricular activities impacted the acculturation of first-generation adolescent males. Specifically, the project focused on the influence of organized high school soccer on the development of first-generation adolescent Korean American males. Eight adolescent participants, ranging in age from fourteen…

  14. Parvalbumin-Positive Inhibitory Interneurons Oppose Propagation But Favor Generation of Focal Epileptiform Activity.

    PubMed

    Sessolo, Michele; Marcon, Iacopo; Bovetti, Serena; Losi, Gabriele; Cammarota, Mario; Ratto, Gian Michele; Fellin, Tommaso; Carmignoto, Giorgio

    2015-07-01

    Parvalbumin (Pv)-positive inhibitory interneurons effectively control network excitability, and their optogenetic activation has been reported to block epileptic seizures. An intense activity in GABAergic interneurons, including Pv interneurons, before seizures has been described in different experimental models of epilepsy, raising the hypothesis that an increased GABAergic inhibitory signal may, under certain conditions, initiate seizures. It is therefore unclear whether the activity of Pv interneurons enhances or opposes epileptiform activities. Here we use a mouse cortical slice model of focal epilepsy in which the epileptogenic focus can be identified and the role of Pv interneurons in the generation and propagation of seizure-like ictal events is accurately analyzed by a combination of optogenetic, electrophysiological, and imaging techniques. We found that a selective activation of Pv interneurons at the focus failed to block ictal generation and induced postinhibitory rebound spiking in pyramidal neurons, enhancing neuronal synchrony and promoting ictal generation. In contrast, a selective activation of Pv interneurons distant from the focus blocked ictal propagation and shortened ictal duration at the focus. We revealed that the reduced ictal duration was a direct consequence of the ictal propagation block, probably by preventing newly generated afterdischarges to travel backwards to the original focus of ictal initiation. Similar results were obtained upon individual Pv interneuron activation by intracellular depolarizing current pulses. The functional dichotomy of Pv interneurons here described opens new perspectives to our understanding of how local inhibitory circuits govern generation and spread of focal epileptiform activities.

  15. Enhanced colonic nitric oxide generation and nitric oxide synthase activity in ulcerative colitis and Crohn's disease.

    PubMed Central

    Rachmilewitz, D; Stamler, J S; Bachwich, D; Karmeli, F; Ackerman, Z; Podolsky, D K

    1995-01-01

    Recent studies have suggested that nitric oxide (NO.), the product of nitric oxide synthase in inflammatory cells, may play a part in tissue injury and inflammation through its oxidative metabolism. In this study the colonic generation of oxides of nitrogen (NOx) and nitric oxide synthase activity was determined in ulcerative colitis and Crohn's disease. Colonic biopsy specimens were obtained from inflammatory bowel disease patients and from normal controls. Mucosal explants were cultured in vitro for 24 hours and NOx generation was determined. Nitric oxide synthase activity was monitored by the conversion of [3H]-L-arginine to citrulline. Median NOx generation by inflamed colonic mucosa of patients with active ulcerative colitis and Crohn's colitis was 4.2- and 8.1-fold respectively higher than that by normal human colonic mucosa. In ulcerative colitis and Crohn's colitis nitric oxide synthase activity was 10.0- and 3.8-fold respectively higher than in normal subjects. Colonic NOx generation is significantly decreased by methylprednisolone and ketotifen. The decrease in NOx generation by cultured colonic mucosa induced by methylprednisolone suggests that NO synthase activity is induced during the culture and the steroid effect may contribute to its therapeutic effect. Enhanced colonic NOx generation by stimulated nitric oxide synthase activity in ulcerative colitis and Crohn's disease may contribute to tissue injury. PMID:7541008

  16. Electrostatic power generation using carbon-activated cotton thread on textile

    NASA Astrophysics Data System (ADS)

    Kim, Baek Hyun; Barnhart, Benjamin S.; Kwon, Jae W.

    2015-12-01

    This paper describes a novel thread-shaped power generator which can be incorporated into cloth. A carbon-activated cotton thread is utilized for harvesting electrostatic energy from environment using contact and friction electrifications. A core of cotton thread was treated with carbon black nano particles to provide conductivity, and then encapsulated with a thin layer of polydimethylsiloxane for stability and protection. Electrostatic charges have been collected from carbon-activated threads stitched on pieces of textiles by repeated rubbing and tapping with a ploytetrafluoethylene sheet. An average open-circuit voltage of approximately -60.9 V has been generated from the thread-shaped generator with rubbing mode.

  17. Oncogenic S1P signalling in EBV-associated nasopharyngeal carcinoma activates AKT and promotes cell migration through S1P receptor 3.

    PubMed

    Lee, Hui Min; Lo, Kwok-Wai; Wei, Wenbin; Tsao, Sai Wah; Chung, Grace Tin Yun; Ibrahim, Maha Hafez; Dawson, Christopher W; Murray, Paul G; Paterson, Ian C; Yap, Lee Fah

    2017-02-27

    Undifferentiated nasopharyngeal carcinoma (NPC) is a cancer with high metastatic potential that is consistently associated with Epstein-Barr virus (EBV) infection. In this study, we have investigated the functional contribution of sphingosine-1-phosphate (S1P) signalling to the pathogenesis of NPC. We show that EBV infection or ectopic expression of the EBV-encoded latent genes (EBNA1, LMP1 and LMP2A) can up-regulate sphingosine kinase 1 (SPHK1), the key enzyme that produces S1P, in NPC cell lines. Exogenous addition of S1P promotes the migration of NPC cells through the activation of AKT; shRNA knockdown of SPHK1 resulted in a reduction in the levels of activated AKT and inhibition of cell migration. We also show that S1P receptor 3 (S1PR3) mRNA is over-expressed in EBV-positive NPC patient-derived xenografts and a subset of primary NPC tissues, and that knockdown of S1PR3 suppressed the activation of AKT and the S1P-induced migration of NPC cells. Taken together, our data point to a central role for EBV in mediating the oncogenic effects of S1P in NPC and identify S1P signalling as a potential therapeutic target in this disease.

  18. Demonstration of the serum antibody to Epstein-Barr virus specific DNA polymerased (EBV-DP) from patients with nasopharyngeal carcinoma (NPC)

    SciTech Connect

    Tan, R.S.; Li, J.S.; Grill, S.; Nutter, L.M.; Cheng, Y.C.

    1986-03-05

    Raji cells, an EBV genome carrying and nonproducer cell line, treated with tetradecanoyl-phorbol-13-acetate (TPA) and n-butyrate could induce a special DNA polymerase which has properties that are similar to the EBV-DP induced by TPA in P/sub 3/HR-I cells, an EBV producer cell line. Since EBV was found to have a strong association with NPC, and antibodies against EBV proteins or enzymes were found in high levels in sera from these patients, the possible presence of serum antibody against EBV-DP was examined. The serum titer of antibody to EBV-DP was found to have 190 +/- 84 units/ml serum (mean +/- S.D.) in 48 sera from patients with NPC. The titer in 52 healthy donors was 31.4 +/- 28 unit/ml serum (p < 0.01). The antibody was found to be associated with the IgG but not the IgA fraction. The antibody titers against EBV-DP were not correlated with the titer against EBV-DNase or VCA-IgA. Whether the antibody observed is against an EBV-DP core protein or its stimulating protein, as demonstrated by this laboratory previously, is still being investigated. This study demonstrated the high frequency and high titer of antibody against EBV-DP in serum from patients with NPC, and suggested the potential of utilizing this antibody titer to compliment other methods for the early diagnosis or prognosis of NPC.

  19. A Curriculum Activities Guide to Electric Power Generation and the Environment.

    ERIC Educational Resources Information Center

    Tully, Randolph R., Jr., Ed.

    This guide was developed by teachers involved in a workshop on "Electric Power Generation and the Environment." Activity topics are: (1) Energy and the Consumer; (2) Energy and Water Pollution; and (3) Energy and Air Pollution. Within these topics, the activities are classified as awareness level, transitional level, or operational…

  20. Exploring Preferences of Mentoring Activities among Generational Groups of Registered Nurses in Florida

    ERIC Educational Resources Information Center

    Posey-Goodwin, Patricia Ann

    2013-01-01

    The purpose of this study was to explore differences in perceptions of mentoring activities from four generations of registered nurses in Florida, using the Alleman Mentoring Activities Questionnaire ® (AMAQ ®). Statistical procedures of analysis of variance (ANOVA) were employed to explore differences among 65 registered nurses in Florida from…

  1. Contradictions between the Virtual and Physical High School Classroom: A Third-Generation Activity Theory Perspective

    ERIC Educational Resources Information Center

    Murphy, Elizabeth; Manzanares, Maria A. Rodriguez

    2008-01-01

    This paper uses a third-generation Activity Theory perspective to gain insight into the contradictions between the activity systems of the physical and virtual high school classroom from the perspective of teachers who had transitioned from one system to the other. Data collection relied on semi-structured interviews conducted with e-teachers as…

  2. An Evaluation of a Wide Range of Job-Generating Activities for Rural Counties.

    ERIC Educational Resources Information Center

    Finsterbusch, Kurt; And Others

    1992-01-01

    Examines the job-generating activities in 15 rural counties in Maryland through 175 interviews and field work. Those ranking high included industrial park development, economic development activities, and tourism. Special financial arrangements for relocating and new businesses also received high marks. Includes 48 references. (JOW)

  3. Analysing Third Generation Activity Systems: Labour-Power, Subject Position and Personal Transformation

    ERIC Educational Resources Information Center

    Daniels, Harry; Warmington, Paul

    2007-01-01

    Purpose: The purpose of this paper is to describe how Engestrom's "third generation" activity theory, with its emphasis on developing conceptual tools to understand dialogues, multiple perspectives and networks of interacting activity systems, has informed research into professional learning in multiagency service settings in England.…

  4. Human Platelets Utilize Cycloxygenase-1 to Generate Dioxolane A3, a Neutrophil-activating Eicosanoid*

    PubMed Central

    Hinz, Christine; Aldrovandi, Maceler; Uhlson, Charis; Marnett, Lawrence J.; Longhurst, Hilary J.; Warner, Timothy D.; Alam, Saydul; Slatter, David A.; Lauder, Sarah N.; Allen-Redpath, Keith; Collins, Peter W.; Murphy, Robert C.; Thomas, Christopher P.; O'Donnell, Valerie B.

    2016-01-01

    Eicosanoids are important mediators of fever, pain, and inflammation that modulate cell signaling during acute and chronic disease. We show by using lipidomics that thrombin-activated human platelets generate a new type of eicosanoid that both stimulates and primes human neutrophil integrin (Mac-1) expression, in response to formylmethionylleucylphenylalanine. Detailed characterization proposes a dioxolane structure, 8-hydroxy-9,11-dioxolane eicosatetraenoic acid (dioxolane A3, DXA3). The lipid is generated in nanogram amounts by platelets from endogenous arachidonate during physiological activation, with inhibition by aspirin in vitro or in vivo, implicating cyclooxygenase-1 (COX). Pharmacological and genetic studies on human/murine platelets revealed that DXA3 formation requires protease-activated receptors 1 and 4, cytosolic phospholipase A2 (cPLA2), Src tyrosine kinases, p38 MAPK, phospholipase C, and intracellular calcium. From data generated by purified COX isoforms and chemical oxidation, we propose that DXA3 is generated by release of an intermediate from the active site followed by oxygenation at C8. In summary, a new neutrophil-activating platelet-derived lipid generated by COX-1 is presented that can activate or prime human neutrophils, suggesting a role in innate immunity and acute inflammation. PMID:27129261

  5. Beyond participation: the association between school extracurricular activities and involvement in violence across generations of immigration.

    PubMed

    Jiang, Xin; Peterson, Ruth D

    2012-03-01

    Participation in extracurricular activities is purported to protect the broad spectrum of youth from a host of behavioral risks. Yet, empirical research on the extent to which this assumption holds for involvement in violence by immigrant youth is limited. Thus, using data for 13,236 (51.8% female) adolescents from the National Longitudinal Study of Adolescent Health, this study explores how the relationship between extracurricular activities and youth violence varies by type of extracurricular activity profile (sports alone, non-sports alone, and a combination of sports and non-sports) and by generations of immigration (first, second, and third-plus). The sample is composed of 9.3% (n = 1,233) first-generation youth, 15.7% (n = 2,080) second generation, and 74.9% (n = 9,923) third-plus generation. The results reveal that adolescents from the third-plus generation (i.e., non-immigrant youth) who participate in non-sports alone or sports plus non-sports have lower odds of involvement in violence than adolescents from the same generation who do not participate in extracurricular activities. However, for first- and second-generation adolescents, participation in extracurricular activities is associated with higher rather than lower odds of violence compared to their non-participating counterparts. These findings challenge the viewpoint that participation in mainstream extracurricular activities as afforded by US schools is equally beneficial for all youth. They also call for additional research that explores why immigrant youth are less likely than non-immigrant youth to gain violence-reducing benefits when they participate in extracurricular activities.

  6. Estimation of the activity generated by neutron activation in control rods of a BWR.

    PubMed

    Ródenas, José; Gallardo, Sergio; Abarca, Agustín; Juan, Violeta

    2010-01-01

    Control rods are activated by neutron reactions into the reactor. The activation is produced mainly in stainless steel and its impurities. The dose produced by this activity is not important inside the reactor, but it has to be taken into account when the rod is withdrawn from the reactor. Activation reactions produced have been modelled by the MCNP5 code based on the Monte Carlo method. The code gives the number of reactions that can be converted into activity.

  7. Future of low specific activity molybdenum-99/technetium-99m generator.

    PubMed

    Mushtaq, A

    2012-10-01

    In last few years, the shortage of molybdenum-99 (99Mo) was felt in the developed and developing countries hospitals, where diagnostic nuclear medicine is practiced. To overcome the shortage of 99Mo various routes of its production by accelerators and reactors generating low and high specific activity products have been planned. High specific activity 99Mo obtained by fission of uranium-235 (235U) has completely dominated in the manufacturing of technetium-99m (99mTc) generators in last 3-4 decades, but due to proliferation and dirty bomb, issues non fission routes of 99Mo production are emphasized. Future of low specific activity 99Mo is discussed.

  8. Entropy Generation and Human Aging: Lifespan Entropy and Effect of Physical Activity Level

    NASA Astrophysics Data System (ADS)

    Silva, Carlos; Annamalai, Kalyan

    2008-06-01

    The first and second laws of thermodynamics were applied to biochemical reactions typical of human metabolism. An open-system model was used for a human body. Energy conservation, availability and entropy balances were performed to obtain the entropy generated for the main food components. Quantitative results for entropy generation were obtained as a function of age using the databases from the U.S. Food and Nutrition Board (FNB) and Centers for Disease Control and Prevention (CDC), which provide energy requirements and food intake composition as a function of age, weight and stature. Numerical integration was performed through human lifespan for different levels of physical activity. Results were presented and analyzed. Entropy generated over the lifespan of average individuals (natural death) was found to be 11,404 kJ/ºK per kg of body mass with a rate of generation three times higher on infants than on the elderly. The entropy generated predicts a life span of 73.78 and 81.61 years for the average U.S. male and female individuals respectively, which are values that closely match the average lifespan from statistics (74.63 and 80.36 years). From the analysis of the effect of different activity levels, it is shown that entropy generated increases with physical activity, suggesting that exercise should be kept to a “healthy minimum” if entropy generation is to be minimized.

  9. Generation, Language, Body Mass Index, and Activity Patterns in Hispanic Children

    PubMed Central

    Taverno, Sharon E.; Rollins, Brandi Y.; Francis, Lori A.

    2010-01-01

    Background The acculturation hypothesis proposes an overall disadvantage in health outcomes for Hispanic immigrants with more time spent living in the U.S., but little is known about how generational status and language may influence Hispanic children’s relative weight and activity patterns. Purpose The association between generation and language was investigated with relative weight (BMI z-scores), physical activity, screen time, and participation in extracurricular activities (e.g., sports, clubs) in a U.S.-based, nationally representative sample of Hispanic children. Methods Participants included 2,012 Hispanic children aged 6–11 years from the cross-sectional, 2003 National Survey of Children’s Health. Children were grouped according to generational status (1st, 2nd or 3rd), and the primary language spoken in the home (English vs non-English). Primary analyses included adjusted logistic and multinomial logistic regression to examine the relationships among variables; all analyses were conducted between 2008 and 2009. Results Compared to 3rd generation, English speakers, 1st and 2nd generation, non-English speakers were over two times more likely to be obese. Moreover, 1st generation, non-English speakers were half as likely to engage in regular physical activity and sports. Both 1st and 2nd generation, non-English speakers were less likely to participate in clubs compared to 2nd and 3rd generation, English speakers. Overall, all non–English speaking groups reported less screen time compared to 3rd generation, English speakers. Conclusions The hypothesis that Hispanics lose their health protection with more time spent in the U.S. was not supported in this sample of Hispanic children. PMID:20117570

  10. Active Oxygen Species Generator by Low Pressure Silent Discharge and its Application to Water Treatment

    NASA Astrophysics Data System (ADS)

    Tanaka, Masaaki; Ikeda, Akira; Tanimura, Yasuhiro; Ohta, Koji; Yoshiyasu, Hajimu

    We have proposed the new water treatment using the active oxygen species such as an atomic oxygen with the oxidation power that is stronger than ozone. Based on the results of simulations we designed the silent discharge type active oxygen generator with a water ejector, which is operated on the discharge conditions of low pressure of 6.6kPa. and high temperature of about 200°C. The experimental results are as follows. (1) The yield of the active oxygen increases with the increase of the discharge tube temperature and the decrease of the gas pressure. (2) The life time of active oxygen is tens msec. (3) The active oxygen oxidizes efficiently the formic acid compared with ozone. It is assumed from these results that the active oxygen species having a strong oxidation power is generated.

  11. Fiscal Year 2014 Annual Report on BNLs Next Generation Safeguards Initiative Human Capital Development Activities

    SciTech Connect

    Pepper, Susan E.

    2014-10-10

    Brookhaven National Laboratory’s (BNL’s) Nonproliferation and National Security Department contributes to the National Nuclear Security Administration Office of Nonproliferation and International Security Next Generation Safeguards Initiative (NGSI) through university engagement, safeguards internships, safeguards courses, professional development, recruitment, and other activities aimed at ensuring the next generation of international safeguards professionals is adequately prepared to support the U.S. safeguards mission. This report is a summary of BNL s work under the NGSI program in Fiscal Year 2014.

  12. Inference generation during text comprehension by adults with right hemisphere brain damage: activation failure versus multiple activation.

    PubMed

    Tompkins, Connie A; Fassbinder, Wiltrud; Lehman Blake, Margaret; Baumgaertner, Annette; Jayaram, Nandini

    2004-12-01

    Evidence conflicts as to whether adults with right hemisphere brain damage (RHD) generate inferences during text comprehension. M. Beeman (1993) reported that adults with RHD fail to activate the lexical-semantic bases of routine bridging inferences, which are necessary for comprehension. But other evidence indicates that adults with RHD activate multiple interpretations in various comprehension domains. In addition, the activation of contextually inappropriate interpretations is prolonged for many adults with RHD and predicts poor discourse comprehension. This study contrasted Beeman's activation failure hypothesis with the prediction that adults with RHD would generate multiple interpretations in text comprehension. The relation between activation of textually incompatible inferences and discourse comprehension was also investigated for this group. Thirty-seven adults with RHD and 34 without brain damage listened to brief narratives that required a bridging inference (BI) to integrate the text-final sentence. This final sentence, when isolated from its text, was strongly biased toward a contextually incompatible alternate interpretation (AI). Auditory phoneme strings were presented for lexical decision immediately after each text's initial and final sentence. Adults with RHD were both faster and more accurate in making lexical decisions to BI-related target words in final-sentence position than in initial-sentence position. Thus, contrary to the activation failure hypothesis, adults with RHD generated the lexical-semantic foundations of BIs where they were required by the text. AI generation was evident in accuracy data as well, but not in response time data. This result is partially consistent with the multiple activation view. Finally, greater activation for contextually incompatible interpretations was associated with poorer discourse comprehension performance by adults with RHD.

  13. Multiscale Aspects of Generation of High-Gamma Activity during Seizures in Human Neocortex123

    PubMed Central

    Marcuccilli, Charles J.; Ben-Mabrouk, Faiza; Lew, Sean M.; Goodman, Robert R.; McKhann, Guy M.; Frim, David M.; Kohrman, Michael H.; Schevon, Catherine A.; van Drongelen, Wim

    2016-01-01

    High-gamma (HG; 80-150 Hz) activity in macroscopic clinical records is considered a marker for critical brain regions involved in seizure initiation; it is correlated with pathological multiunit firing during neocortical seizures in the seizure core, an area identified by correlated multiunit spiking and low frequency seizure activity. However, the effects of the spatiotemporal dynamics of seizure on HG power generation are not well understood. Here, we studied HG generation and propagation, using a three-step, multiscale signal analysis and modeling approach. First, we analyzed concurrent neuronal and microscopic network HG activity in neocortical slices from seven intractable epilepsy patients. We found HG activity in these networks, especially when neurons displayed paroxysmal depolarization shifts and network activity was highly synchronized. Second, we examined HG activity acquired with microelectrode arrays recorded during human seizures (n = 8). We confirmed the presence of synchronized HG power across microelectrode records and the macroscale, both specifically associated with the core region of the seizure. Third, we used volume conduction-based modeling to relate HG activity and network synchrony at different network scales. We showed that local HG oscillations require high levels of synchrony to cross scales, and that this requirement is met at the microscopic scale, but not within macroscopic networks. Instead, we present evidence that HG power at the macroscale may result from harmonics of ongoing seizure activity. Ictal HG power marks the seizure core, but the generating mechanism can differ across spatial scales. PMID:27257623

  14. The generation of rhythmic activity in dissociated cultures of rat spinal cord.

    PubMed

    Streit, J; Tscherter, A; Heuschkel, M O; Renaud, P

    2001-07-01

    Locomotion in vertebrates is controlled by central pattern generators in the spinal cord. The roles of specific network architecture and neuronal properties in rhythm generation by such spinal networks are not fully understood. We have used multisite recording from dissociated cultures of embryonic rat spinal cord grown on multielectrode arrays to investigate the patterns of spontaneous activity in randomised spinal networks. We were able to induce similar patterns of rhythmic activity in dissociated cultures as in slice cultures, although not with the same reliability and not always with the same protocols. The most reliable rhythmic activity was induced when a partial disinhibition of the network was combined with an increase in neuronal excitability, suggesting that both recurrent synaptic excitation and neuronal excitability contribute to rhythmogenesis. During rhythmic activity, bursts started at several sites and propagated in variable ways. However, the predominant propagation patterns were independent of the protocol used to induce rhythmic activity. When synaptic transmission was blocked by CNQX, APV, strychnine and bicuculline, asynchronous low-rate activity persisted at approximately 50% of the electrodes and approximately 70% of the sites of burst initiation. Following the bursts, the activity in the interval was transiently suppressed below the level of intrinsic activity. The degree of suppression was proportional to the amount of activity in the preceding burst. From these findings we conclude that rhythmic activity in spinal cultures is controlled by the interplay of intrinsic neuronal activity and recurrent excitation in neuronal networks without the need for a specific architecture.

  15. Activating Photodynamic Therapy in vitro with Cerenkov Radiation Generated from Yttrium-90

    PubMed Central

    Hartl, Brad A.; Hirschberg, Henry; Marcu, Laura; Cherry, Simon R.

    2017-01-01

    The translation of photodynamic therapy (PDT) to the clinical setting has primarily been limited to easily accessible and/or superficial diseases, for which traditional light delivery can be performed noninvasively. Cerenkov radiation, as generated from medically relevant radionuclides, has been suggested as a means to deliver light to deeper tissues noninvasively to overcome this depth limitation. This article investigates the utility of Cerenkov radiation, as generated from the radionuclide yttrium-90, for activating the PDT process using clinically approved aminolevulinic acid at 1.0 mm and also the more efficient porphyrin-based photosensitizer mesotetraphenylporphine with two sulfonate groups on adjacent phenyl rings (TPPS2a) at 1.2 μM. Experiments were conducted with monolayer cultured glioma and breast tumor cell lines. Although aminolevulinic acid proved to be ineffective for generating a therapeutic effect at all but the highest activity levels, TPPS2a produced at least a 20% therapeutic effect at activities ranging from 6 to 60 μCi/well for the C6 glioma cell line. Importantly, these results demonstrate for the first time, to our knowledge, that Cerenkov radiation generated from a radionuclide can be used to activate PDT using clinically relevant photosensitizers. These results therefore provide evidence that it may be possible to generate a phototherapeutic effect in vivo using Cerenkov radiation and clinically relevant photosensitizers. PMID:27481495

  16. Prothrombin-activator (thromboplastin) generation in the blood of water snake (Natrix piscator).

    PubMed

    Srivastava, V M; Dube, B; Dube, R K; Agarwal, G P

    1981-12-23

    The generation of prothrombin-activator (thromboplastin) in water snake (Natrix piscator) is clearly delayed, compared to a mammalian system, but the final activity is well comparable to that in man, when homologous sources of "phospholipid" (erythrocyte-lysate) and of substrate plasma are employed in one stage "thromboplastin generation test". The use of heterologous source of either of the above reagents resulted in significantly longer clotting times; hence the need for homologous source of above reagents in the test is emphasized for comparative studies on animal haemostasis.

  17. On-Orbit Checkout and Activation of the ISS Oxygen Generation System

    NASA Technical Reports Server (NTRS)

    Bagdigian, Robert M.; Prokhorov, Kimberlee S.

    2007-01-01

    NASA has developed and; deployed an Oxygen Generation System (OGS) into the Destiny Module of the International Space Station (ISS). The major. assembly; included in this system is the Oxygen Generator Assembly. (OGA) which was developed under NASA contract by Hamilton Sundstrand Space Systems International (HSSSI), Inc. This paper summarizes the installation of the system into the Destiny Module, its initial checkout and periodic preventative maintenance activities, and its operational activation. Trade studies and analyses that were conducted with the goal of mitigating on-orbit operational risks are also discussed.

  18. Energy deposition in the earth's atmosphere due to impact of solar activity-generated disturbances

    NASA Technical Reports Server (NTRS)

    Wu, S. T.; Kan, L. C.; Tandberg-Hanssen, E.; Dryer, M.

    1979-01-01

    Energy deposition in and dynamic responses of the terrestrial atmosphere to solar flare-generated shocks and other physical processes - such as particle precipitation and local heating - are investigated self-consistently in the context of hydrodynamics, the problem being treated as an initial boundary-value problem. It is extremely difficult to construct a general model for the line solar activity-magnetosphere-atmosphere; however, a limited model for this link is possible. The paper describes such a model, and presents some results on energy deposition into the earth's atmosphere due to solar activity-generated disturbances. Results from the present calculations are presented and discussed.

  19. Activation of photodynamic therapy in vitro with Cerenkov luminescence generated from Yttrium-90 (Conference Presentation)

    NASA Astrophysics Data System (ADS)

    Hartl, Brad A.; Hirschberg, Henry; Marcu, Laura; Cherry, Simon R.

    2016-03-01

    Translation of photodynamic therapy to the clinical setting has primarily been limited to easily accessible and/or superficial diseases where traditional light delivery can be performed noninvasively. Cerenkov luminescence, as generated from medically relevant radionuclides, has been suggested as a means to deliver light to deeper tissues noninvasively in order to overcome this depth limitation. We report on the use of Cerenkov luminescence generated from Yttrium-90 as a means to active the photodynamic therapy process in monolayer tumor cell cultures. The current study investigates the utility of Cerenkov luminescence for activating both the clinically relevant aminolevulinic acid at 1.0 mM and also the more efficient photosensitizer TPPS2a at 1.2 µM. Cells were incubated with aminolevulinic acid for 6 hours prior to radionuclide addition, as well as additional daily treatments for three days. TPPS2a was delivered as a single treatment with an 18 hour incubation time before radionuclide addition. Experiments were completed for both C6 glioma cells and MDA-MB-231 breast tumor cells. Although aminolevulinic acid proved ineffective for generating a therapeutic effect at any activity for either cell line, TPPS2a produced at least a 20% therapeutic effect at activities ranging from 6 to 60 µCi/well for the C6 cell line. Current results demonstrate that it may be possible to generate a therapeutic effect in vivo using Cerenkov luminescence to activate the photodynamic therapy process with clinically relevant photosensitizers.

  20. Drawing as a Generative Activity and Drawing as a Prognostic Activity

    ERIC Educational Resources Information Center

    Schwamborn, Annett; Mayer, Richard E.; Thillmann, Hubertina; Leopold, Claudia; Leutner, Detlev

    2010-01-01

    In this study, 9th-grade students (N = 196) with a mean age of 14.7 years read a scientific text explaining the chemical process of doing laundry with soap and water and then took 3 tests. Students who were instructed to generate drawings during learning scored higher than students who only read on subsequent tests of transfer (d = 0.91),…

  1. Sensory-induced activation of pattern generators in the absence of supraspinal control.

    PubMed

    Lev-Tov, A; Etlin, A; Blivis, D

    2010-06-01

    Sacrocaudal afferent (SCA) stimulation is used in this work to study neural pathways involved in sensory-activation of central pattern generators (CPGs) in the isolated spinal cord of the neonatal rat. Surgical manipulations of the white matter funiculi and confocal imaging of back-labeled funicular pathways suggest that the CPGs are activated during SCA stimulation by crossed and uncrossed multifunicular projections of sacral neurons and that activation of short projecting proprioneurons is sufficient for the generation of the rhythm by SCA stimulation. The versatile organization of the pathways involved in the SCA-induced rhythm makes it a potent and durable activator of the CPGs in the absence of descending control from the brain. The significance of our findings and their potential clinical use are discussed.

  2. Hydrogel design of experiments methodology to optimize hydrogel for iPSC-NPC culture.

    PubMed

    Lam, Jonathan; Carmichael, S Thomas; Lowry, William E; Segura, Tatiana

    2015-03-11

    Bioactive signals can be incorporated in hydrogels to direct encapsulated cell behavior. Design of experiments methodology methodically varies the signals systematically to determine the individual and combinatorial effects of each factor on cell activity. Using this approach enables the optimization of three ligands concentrations (RGD, YIGSR, IKVAV) for the survival and differentiation of neural progenitor cells.

  3. Extracellular serglycin upregulates the CD44 receptor in an autocrine manner to maintain self-renewal in nasopharyngeal carcinoma cells by reciprocally activating the MAPK/β-catenin axis

    PubMed Central

    Chu, Qiaoqiao; Huang, Hongbing; Huang, Tiejun; Cao, Li; Peng, Lixia; Shi, Simei; Zheng, Lisheng; Xu, Liang; Zhang, Shijun; Huang, Jialing; Li, Xinjian; Qian, Chaonan; Huang, Bijun

    2016-01-01

    Serglycin is a proteoglycan that was first found to be secreted by hematopoietic cells. As an extracellular matrix (ECM) component, serglycin promotes nasopharyngeal carcinoma (NPC) metastasis and serves as an independent, unfavorable NPC prognostic indicator. The detailed mechanism underlying the roles of serglycin in cancer progression remains to be clarified. Here, we report that serglycin knockdown in NPC cells inhibited cell sphere formation and tumor seeding abilities. Serglycin downregulation enhanced high-metastasis NPC cell sensitivity to chemotherapy. It has been reported that serglycin is a novel ligand for the stem cell marker CD44. Interestingly, we found a positive correlation between serglycin expression and CD44 in nasopharyngeal tissues and NPC cell lines. Further study revealed that CD44 was an ERK-dependent downstream effector of serglycin signaling, and serglycin activated the MAPK/β-catenin axis to induce CD44 receptor expression in a positive feedback loop. Taken together, our novel findings suggest that ECM serglycin upregulated CD44 receptor expression to maintain NPC stemness by interacting with CD44 and activating the MAPK/β-catenin pathway, resulting in NPC cell chemoresistance. These findings suggest that the intervention of serglycin/CD44 axis and downstream signaling pathway is a rational strategy for targeting NPC cancer stem cell therapy. PMID:27809309

  4. Self-generation of dissipative solitons in magnonic quasicrystal active ring resonator

    NASA Astrophysics Data System (ADS)

    Grishin, S. V.; Beginin, E. N.; Morozova, M. A.; Sharaevskii, Yu. P.; Nikitov, S. A.

    2014-02-01

    Self-generation of dissipative solitons in the magnonic quasicrystal (MQC) active ring resonator is studied theoretically and experimentally. The developed magnonic crystal has quasiperiodic Fibonacci type structure. Frequency selectivity of the MQC together with the parametric three-wave decay of magnetostatic surface spin wave (MSSW) leads to the dissipative soliton self-generation. The transfer matrix method is used to describe MQC transmission responses. Besides, the model of MQC active ring resonator is suggested. The model includes three coupled differential equations describing the parametric decay of MSSW and two differential equations of linear oscillators describing the frequency selectivity of MQC. Numerical simulation results of dissipative soliton self-generation are in a fair agreement with experimental data.

  5. Self-generation of dissipative solitons in magnonic quasicrystal active ring resonator

    SciTech Connect

    Grishin, S. V. Beginin, E. N.; Morozova, M. A.; Sharaevskii, Yu. P.; Nikitov, S. A.

    2014-02-07

    Self-generation of dissipative solitons in the magnonic quasicrystal (MQC) active ring resonator is studied theoretically and experimentally. The developed magnonic crystal has quasiperiodic Fibonacci type structure. Frequency selectivity of the MQC together with the parametric three-wave decay of magnetostatic surface spin wave (MSSW) leads to the dissipative soliton self-generation. The transfer matrix method is used to describe MQC transmission responses. Besides, the model of MQC active ring resonator is suggested. The model includes three coupled differential equations describing the parametric decay of MSSW and two differential equations of linear oscillators describing the frequency selectivity of MQC. Numerical simulation results of dissipative soliton self-generation are in a fair agreement with experimental data.

  6. Recent activities on CCMHD power generation studies with Fuji-1 blow-down facility

    SciTech Connect

    Okuno, Y.; Okamura, T.; Yoshikawa, K.

    1995-12-31

    Recent activities on experimental studies with closed cycle MBD {open_quotes}Fuji-1{open_quotes} blow-down facility at Tokyo Institute of Technology are presented. A new disk generator ( Disk-F4 ) has been installed and a new seed injection system has been introduced from IVTAN (Institute of High Temperature in Russia) in 1994.The design concept of the new generator channel is focused mainly on the reliability of high power generation. The Mach number at the generator inlet and the thermal input are increased up to {approximately}2.8 and {approximately}3.0 MW, respectively. In the new seeding system, a melted seed material is pushed by a piston dozator, instead of gas-pressure-driven in the previous system. The controllability of seed fraction is markedly improved, and the large fluctuation as has been observed previously is diminished, although the spatial non-uniformity of seed fraction which could occur in the all previous experiments is observed. In the power generation experiments with the new components, the maximum power output of 502 kW and the enthalpy extraction ratio of 16.7% have been obtained. These values at the present stage are lower than the maximum values previously achieved in the facility. However, the reliable high power generation can be expected for the new generator.

  7. Developing Students' Listening Metacognitive Strategies Using Online Videotext Self-Dictation-Generation Learning Activity

    ERIC Educational Resources Information Center

    Chang, Ching; Chang, Chih-Kai

    2014-01-01

    The study is based on the use of a flexible learning framework to help students improve information processes underlying strategy instruction in EFL listening. By exploiting the online videotext self-dictation-generation (video-SDG) learning activity implemented on the YouTube caption manager platform, the learning cycle was emphasized to promote…

  8. 7 CFR 1484.38 - Can a Cooperator keep proceeds generated from an activity?

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 7 Agriculture 10 2013-01-01 2013-01-01 false Can a Cooperator keep proceeds generated from an activity? 1484.38 Section 1484.38 Agriculture Regulations of the Department of Agriculture (Continued) COMMODITY CREDIT CORPORATION, DEPARTMENT OF AGRICULTURE EXPORT PROGRAMS PROGRAMS TO HELP DEVELOP...

  9. 7 CFR 1484.38 - Can a Cooperator keep proceeds generated from an activity?

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 7 Agriculture 10 2014-01-01 2014-01-01 false Can a Cooperator keep proceeds generated from an activity? 1484.38 Section 1484.38 Agriculture Regulations of the Department of Agriculture (Continued) COMMODITY CREDIT CORPORATION, DEPARTMENT OF AGRICULTURE EXPORT PROGRAMS PROGRAMS TO HELP DEVELOP...

  10. 7 CFR 1484.38 - Can a Cooperator keep proceeds generated from an activity?

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 7 Agriculture 10 2012-01-01 2012-01-01 false Can a Cooperator keep proceeds generated from an activity? 1484.38 Section 1484.38 Agriculture Regulations of the Department of Agriculture (Continued) COMMODITY CREDIT CORPORATION, DEPARTMENT OF AGRICULTURE EXPORT PROGRAMS PROGRAMS TO HELP DEVELOP...

  11. Identification of intrinsic catalytic activity for electrochemical reduction of water molecules to generate hydrogen.

    PubMed

    Shinagawa, Tatsuya; Takanabe, Kazuhiro

    2015-06-21

    Insufficient hydronium ion activities at near-neutral pH and under unbuffered conditions induce diffusion-limited currents for hydrogen evolution, followed by a reaction with water molecules to generate hydrogen at elevated potentials. The observed constant current behaviors at near neutral pH reflect the intrinsic electrocatalytic reactivity of the metal electrodes for water reduction.

  12. Fast neutron activation analysis by means of low voltage neutron generator

    NASA Astrophysics Data System (ADS)

    Medhat, M. E.

    A description of D-T neutron generator (NG) is presented. This machine can be used for fast neutron activation analysis applied to determine some selected elements, especially light elements, in different materials. Procedure of neutron flux determination and efficiency calculation is described. Examples of testing some Egyptian natural cosmetics are given.

  13. Stability evaluation and correction of a pulsed neutron generator prompt gamma activation analysis system

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Source output stability is important for accurate measurement in prompt gamma neutron activation. This is especially true when measuring low-concentration elements such as in vivo nitrogen (~2.5% of body weight). We evaluated the stability of the compact DT neutron generator within an in vivo nitrog...

  14. Children with autistic spectrum disorders and speech-generating devices: communication in different activities at home.

    PubMed

    Thunberg, Gunilla; Ahlsén, Elisabeth; Sandberg, Annika Dahlgren

    2007-06-01

    The communication of four children with autistic spectrum disorder was investigated when they were supplied with a speech-generating device (SGD) in three different activities in their home environment: mealtime, story reading and "sharing experiences of the preschool day". An activity based communication analysis, in which collective and individual background factors for the activities were outlined, was used as a basis for the discussion of linguistic coding data derived from video-recordings made before and during SGD intervention. The coded communicative behaviours were engagement in activity, role in turn-taking, communicative form, function and effectiveness. An increase in communicative effectiveness was more noticeable when the SGDs could be used to fulfil goals and roles within the activity. The instruction to the parents to use the SGDs in their communication with the child had an important influence on the activities.

  15. Investigating antimicrobial activity in Rheinheimera sp. due to hydrogen peroxide generated by l-lysine oxidase activity.

    PubMed

    Chen, Wen Ming; Lin, Chang Yi; Sheu, Shih Yi

    2010-05-05

    A greenish yellow pigmented bacterial strain, designated GR5, was recently isolated from a freshwater culture pond for a soft-shell turtle. Phylogenetic analyses based on 16S rRNA gene sequences indicate that strain GR5 belongs to the genus Rheinheimera and its only closest neighbor is the type strain of Rheinheimera texasensis (98.2%). Based on the antibiogram assay, strain GR5 possesses a broad spectrum of antimicrobial activity including Gram-positive and Gram-negative bacteria, yeast, algae, and strain GR5 itself. Strain GR5 can synthesize a macromolecule with antimicrobial activity due to the generation of hydrogen peroxide and this antimicrobial effect can be inhibited by catalase. This antimicrobial activity is active only in complex culture media or chemically defined culture media containing l-lysine. This antimicrobial macromolecule in strain GR5 is shown to be a monomeric protein with a molecular mass of 71kDa and isoelectric point of approximately 3.68. Liquid chromatography-tandem mass spectrometry analyses reveal close similarity of a 19-amino acid fragment derived from this protein to the antibacterial protein, AlpP from the marine bacterium Pseudoalteromonas tunicata D2, and to the antibacterial protein, marinocine, from the marine bacterium Marinomonas mediterranea. This study explores the nature of antimicrobial macromolecule such as l-lysine oxidase. This is the first report on a freshwater bacterium producing antimicrobial activity by generating hydrogen peroxide through its enzymatic activity of l-lysine oxidase.

  16. Synthesis and photodynamic activities of modified benzochlorin derivatives on nasopharyngeal carcinoma cells

    NASA Astrophysics Data System (ADS)

    Chang, Chi K.; Lau, Yan-Kin; Lai, Tat-Shing; Yeung, Lam-Lung; Leung, Wing-Nang; Mak, Nai-Ki

    2004-06-01

    Nasopharyngeal carcinoma (NPC), endemic in Southern China, is ranked as the fourth leading cause of cancer deaths in Hong Kong. In an effort to develop new PDT agents for cancer treatment, with a particular emphasis on the NPC, we have investigated the benzochlorin-type photosensitizers. The chromophore is functionalized with side chains containing polar and/or cationic groups. Synthetic methods have been developed for such purposes; e.g. 5-chlorosulfonyloctaethylbenzochlorin and 5-bromooctaethylbenzochlorin are used as versatile precursors for the preparation of ammonium and amidinium salts. Preliminary in vitro study of the photodynamic activity of the synthetic compounds indicated that the cationic benzochlorin derivatives display significant photocytotoxicity towards NPC cells.

  17. A bibliometric analysis of the development of next generation active nanotechnologies

    NASA Astrophysics Data System (ADS)

    Suominen, Arho; Li, Yin; Youtie, Jan; Shapira, Philip

    2016-09-01

    Delineating the emergence of nanotechnologies that offer new functionalities is an important element in an anticipatory approach to the governance of nanotechnology and its potential impacts. This paper examines the transition to next generation active nanotechnologies which incorporate functions that respond to the environment or systems concepts that combine devices and structures that are dynamic and which may change their states in use. We develop an approach to identifying these active nanotechnologies and then use bibliometric analysis to examine the extent of research papers and patents involving these concepts. We also examine references to environmental, health, and safety concepts in these papers, given that these next generation nanotechnologies are likely to have risk profiles that are different from those of first-generation passive nanomaterials. Our results show a steady growth overall in focus on active nanotechnologies in the research literature and in patents over the study period of 1990-2010. We also find an increase in consideration given to environmental, health, and safety topics. While gaps are highlighted in our understanding of research and innovation in active nanotechnologies, the results suggest that there is beginning to be a shift to active nanotechnologies, with the implication that governance processes need to be conscious of this shift and to prepare for it.

  18. Sphingomyelinase D activity in model membranes: structural effects of in situ generation of ceramide-1-phosphate.

    PubMed

    Stock, Roberto P; Brewer, Jonathan; Wagner, Kerstin; Ramos-Cerrillo, Blanca; Duelund, Lars; Jernshøj, Kit Drescher; Olsen, Lars Folke; Bagatolli, Luis A

    2012-01-01

    The toxicity of Loxosceles spider venom has been attributed to a rare enzyme, sphingomyelinase D, which transforms sphingomyelin to ceramide-1-phosphate. The bases of its inflammatory and dermonecrotic activity, however, remain unclear. In this work the effects of ceramide-1-phosphate on model membranes were studied both by in situ generation of this lipid using a recombinant sphingomyelinase D from the spider Loxosceles laeta and by pre-mixing it with sphingomyelin and cholesterol. The systems of choice were large unilamellar vesicles for bulk studies (enzyme kinetics, fluorescence spectroscopy and dynamic light scattering) and giant unilamellar vesicles for fluorescence microscopy examination using a variety of fluorescent probes. The influence of membrane lateral structure on the kinetics of enzyme activity and the consequences of enzyme activity on the structure of target membranes containing sphingomyelin were examined. The findings indicate that: 1) ceramide-1-phosphate (particularly lauroyl ceramide-1-phosphate) can be incorporated into sphingomyelin bilayers in a concentration-dependent manner and generates coexistence of liquid disordered/solid ordered domains, 2) the activity of sphingomyelinase D is clearly influenced by the supramolecular organization of its substrate in membranes and, 3) in situ ceramide-1-phosphate generation by enzymatic activity profoundly alters the lateral structure and morphology of the target membranes.

  19. Promoting Active Species Generation by Electrochemical Activation in Alkaline Media for Efficient Electrocatalytic Oxygen Evolution in Neutral Media.

    PubMed

    Xu, Kun; Cheng, Han; Liu, Linqi; Lv, Haifeng; Wu, Xiaojun; Wu, Changzheng; Xie, Yi

    2017-01-11

    In this study, by using dicobalt phosphide nanoparticles as precatalysts, we demonstrated that electrochemical activation of metallic precatalysts in alkaline media (comparing with directly electrochemical activation in neutral media) could significantly promote the OER catalysis in neutral media, specifically realizing a 2-fold enhanced activity and meanwhile showing a greatly decreased overpotential of about 100 mV at 10 mA cm(-2). Compared directly with electrochemical activation in neutral media, the electrochemical activation in harsh alkaline media could easily break the strong Co-Co bond and promote active species generation on the surface of metallic Co2P, thus accounting for the enhancement of neutral OER activity, which is also evidenced by HRTEM and the electrochemical double-layer capacitance measurement. The activation of electrochemical oxidation of metallic precatalysts in alkaline media enhanced neutral OER catalysis could also be observed on CoP nanoparticles and Ni2P nanoparticles, suggesting this is a generic strategy. Our work highlights that the activation of electrochemical oxidation of metallic precatalysts in alkaline media would pave new avenues for the design of advanced neutral OER electrocatalysts.

  20. Recognition of human-vehicle interactions in group activities via multi-attributed semantic message generation

    NASA Astrophysics Data System (ADS)

    Elangovan, Vinayak; Shirkhodaie, Amir

    2015-05-01

    Improved Situational awareness is a vital ongoing research effort for the U.S. Homeland Security for the past recent years. Many outdoor anomalous activities involve vehicles as their primary source of transportation to and from the scene where a plot is executed. Analysis of dynamics of Human-Vehicle Interaction (HVI) helps to identify correlated patterns of activities representing potential threats. The objective of this paper is bi-folded. Primarily, we discuss a method for temporal HVI events detection and verification for generation of HVI hypotheses. To effectively recognize HVI events, a Multi-attribute Vehicle Detection and Identification technique (MVDI) for detection and classification of stationary vehicles is presented. Secondly, we describe a method for identification of pertinent anomalous behaviors through analysis of state transitions between two successively detected events. Finally, we present a technique for generation of HVI semantic messages and present our experimental results to demonstrate the effectiveness of semantic messages for discovery of HVI in group activities.

  1. Locomotor-like activity generated by the neonatal mouse spinal cord.

    PubMed

    Bonnot, Agnès; Whelan, Patrick J; Mentis, George Z; O'Donovan, Michael J

    2002-10-01

    This report describes locomotor-like activity generated by the neonatal mouse spinal cord in vitro. We demonstrate that locomotor-like activity can be produced either spontaneously or by a train of stimuli applied to the dorsal roots or in the presence of bath-applied drugs. Calcium imaging of the motoneuron activity generated by a train of dorsal root stimuli revealed a rostrocaudally propagating component of the optical signal in the anterior lumbar (L1-L3) and in the caudal segments (S1-S4). We hypothesize that this spatio-temporal pattern arises from a rostrocaudal gradient of excitability in the relevant segments. Our experiments suggest that left/right reciprocal inhibition and NMDA-mediated oscillations are not essential mechanisms underlying rhythmogenesis in the neonatal mouse cord. Finally, our data are discussed in the context of other models of locomotion in lower and higher vertebrates.

  2. Inhibition of Cholesterol Biosynthesis Reduces γ-Secretase Activity and Amyloid-β Generation.

    PubMed

    Kim, Yoonhee; Kim, Chaeyoung; Jang, Hye Young; Mook-Jung, Inhee

    2016-01-01

    Amyloid-β (Aβ) is one of major molecules contributing to the pathogenesis of Alzheimer's disease (AD). Aβ is derived from amyloid-β protein precursor (AβPP) through sequential cleavages by β- and γ-secretases. Regulation of these components is thought to be an important factor in Aβ generation during the pathogenesis of AD. AβPP, β-secretase, and γ-secretase reside in lipid rafts, where cholesterol regulates the integrity and flexibility of membrane proteins and Aβ is generated. However, the relationship between cholesterol and Aβ generation is controversial. In this study, we aimed to elucidate the direct effects of cholesterol depletion on AβPP processing using AY9944, which blocks the last step of cholesterol biosynthesis and thus minimizes the unknown side effects of upstream inhibitors, such as HMG-CoA reductase inhibitors. Treatment with AY9944 decreased γ-secretase activity and Aβ generation. These results suggested that changes in membrane composition by lowering cholesterol with AY9944 affected γ-secretase activity and Aβ generation, which is associated with AD pathogenesis.

  3. Pattern-dependent Role of NMDA receptors in Action Potential Generation: Consequences on ERK Activation

    PubMed Central

    Zhao, Meilan; Adams, J. Paige

    2005-01-01

    Synaptic long-term potentiation is maintained through gene transcription, but how the nucleus is recruited remains controversial. Activation of extracellular-signal regulated kinases 1 and 2 (ERKs) with synaptic stimulation has been shown to require NMDA receptors (NMDARs), yet stimulation intensities sufficient to recruit action potentials (APs) also appear to be required. This has led us to ask the question whether NMDARs are necessary for AP generation as they relate to ERK activation. To test this, we examined the effects of NMDAR blockade on APs induced with synaptic stimulation using whole-cell current clamp recordings from CA1 pyramidal cells in hippocampal slices. NMDAR antagonists were found to potently inhibit APs generated with 5 and 100 Hz synaptic stimulation. Blockade of APs, and ERK activation, could be overcome with the addition of the GABA-A antagonist bicuculline, indicating that APs are sufficient to activate signals such as ERK in the nucleus and throughout the neuron in the continued presence of NMDAR antagonists. Interestingly, no effects of the NMDAR antagonists were observed when theta-burst stimulation (TBS) was used. This resistance to the antagonists is conferred by temporal summation during the bursts. These results clarify findings from a previous study showing that ERK activation induced with TBS is resistant to APV, in contrast to that induced with 5 Hz or 100 Hz stimulation, which is sensitive. By showing that NMDAR blockade inhibits AP generation, we demonstrate that a major role NMDARs play in cell-wide and nuclear ERK activation is through their contribution to action potential generation. PMID:16049179

  4. Generation of field potentials and modulation of their dynamics through volume integration of cortical activity.

    PubMed

    Kajikawa, Yoshinao; Schroeder, Charles E

    2015-01-01

    Field potentials (FPs) recorded within the brain, often called "local field potentials" (LFPs), are useful measures of net synaptic activity in a neuronal ensemble. However, due to volume conduction, FPs spread beyond regions of underlying synaptic activity, and thus an "LFP" signal may not accurately reflect the temporal patterns of synaptic activity in the immediately surrounding neuron population. To better understand the physiological processes reflected in FPs, we explored the relationship between the FP and its membrane current generators using current source density (CSD) analysis in conjunction with a volume conductor model. The model provides a quantitative description of the spatiotemporal summation of immediate local and more distant membrane currents to produce the FP. By applying the model to FPs in the macaque auditory cortex, we have investigated a critical issue that has broad implications for FP research. We have shown that FP responses in particular cortical layers are differentially susceptible to activity in other layers. Activity in the supragranular layers has the strongest contribution to FPs in other cortical layers, and infragranular FPs are most susceptible to contributions from other layers. To define the physiological processes generating FPs recorded in loci of relatively weak synaptic activity, strong effects produced by synaptic events in the vicinity have to be taken into account. While outlining limitations and caveats inherent to FP measurements, our results also suggest specific peak and frequency band components of FPs can be related to activity in specific cortical layers. These results may help improving the interpretability of FPs.

  5. Protein disulfide isomerase inhibition blocks thrombin generation in humans by interfering with platelet factor V activation

    PubMed Central

    Stopa, Jack D.; Neuberg, Donna; Puligandla, Maneka; Furie, Bruce; Zwicker, Jeffrey I.

    2017-01-01

    BACKGROUND: Protein disulfide isomerase (PDI) is required for thrombus formation. We previously demonstrated that glycosylated quercetin flavonoids such as isoquercetin inhibit PDI activity and thrombus formation in animal models, but whether extracellular PDI represents a viable anticoagulant target in humans and how its inhibition affects blood coagulation remain unknown. METHODS: We evaluated effects of oral administration of isoquercetin on platelet-dependent thrombin generation in healthy subjects and patients with persistently elevated anti-phospholipid antibodies. RESULTS: Following oral administration of 1,000 mg isoquercetin to healthy adults, the measured peak plasma quercetin concentration (9.2 μM) exceeded its IC50 for inhibition of PDI by isoquercetin in vitro (2.5 ± 0.4 μM). Platelet-dependent thrombin generation decreased by 51% in the healthy volunteers compared with baseline (P = 0.0004) and by 64% in the anti-phospholipid antibody cohort (P = 0.015) following isoquercetin ingestion. To understand how PDI affects thrombin generation, we evaluated substrates of PDI identified using an unbiased mechanistic-based substrate trapping approach. These studies identified platelet factor V as a PDI substrate. Isoquercetin blocked both platelet factor Va and thrombin generation with an IC50 of ~5 μM. Inhibition of PDI by isoquercetin ingestion resulted in a 53% decrease in the generation of platelet factor Va (P = 0.001). Isoquercetin-mediated inhibition was reversed with addition of exogenous factor Va. CONCLUSION: These studies show that oral administration of isoquercetin inhibits PDI activity in plasma and diminishes platelet-dependent thrombin generation predominantly by blocking the generation of platelet factor Va. These pharmacodynamic and mechanistic observations represent an important step in the development of a novel class of antithrombotic agents targeting PDI. TRIAL REGISTRATION: Clinicaltrials.gov (NCT01722669) FUNDING: National Heart

  6. Immigrant generation and physical activity among Mexican, Chinese & Filipino adults in the U.S.

    PubMed

    Afable-Munsuz, Aimee; Ponce, Ninez A; Rodriguez, Michael; Perez-Stable, Eliseo J

    2010-06-01

    Migrant studies of physical activity (PA) can provide insight into the prevention of chronic disease. It is unclear, however, whether PA increases or decreases the longer migrants live in their host country. In the US, studies on immigrants' length of residence in the US and PA are inconclusive and many studies do not adequately consider the role of socioeconomic status (SES). Using California data, we examine relationships between immigrant generation and physical activity (PA) among Mexican, Chinese and Filipino adults, who represent the three largest immigrant groups in the US, and the extent to which the relationships are confounded by SES. Data from the 2000 US Census was linked with data on adults 18 years and older from the 2005 California Health Interview Survey. PA was measured in three different domains: leisure time (LTPA), non-leisure time (NLTPA) and any PA. Logistic regression was used to examine whether a wide range of SES factors, measured at the respondent and neighborhood levels, influenced the relationship between immigrant generation and PA in all domains and in different ethnic origin groups. Generation was significantly associated with LTPA among Mexican and Chinese adults and with NLTPA among all 3 ethnic origin groups; however the nature of the relationships varied. After adjusting for individual and neighborhood SES factors, a positive association between generation and LTPA remained among Mexican adults, and negative association between generation and NLTPA remained among Chinese and Filipino adults. These results underscore the importance of comparative studies of immigrant generation and PA and consideration of SES factors to identify pathways linking generation to PA. In the context of increasing rates of chronic disease, the study of transitions in PA among immigrants will continue to be critical to promoting the public health of diverse populations in countries such as the US.

  7. Rostral anterior cingulate activity generates posterior versus anterior theta activity linked to agentic extraversion.

    PubMed

    Chavanon, Mira-Lynn; Wacker, Jan; Stemmler, Gerhard

    2011-06-01

    Recent research using the resting electroencephalogram (EEG) showed that posterior versus anterior theta activity (around 4-8 Hz) is consistently associated with agency, reflecting the dopaminergic core of extraversion (i.e., incentive motivation, positive emotion). Neuroimaging studies using various methodologies and experimental paradigms have converged on the anterior cingulate cortex (ACC) as a neurophysiological correlate of extraversion. The aim of the present study is integrate these lines of research by testing the hypothesis that posterior versus anterior EEG theta is at least partly based on ACC theta activity. Resting EEG data were analyzed in N = 78 healthy, male participants extremely high or low in agentic extraversion (aE). Using the low-resolution electromagnetic tomography algorithm, we localized the sources of aE-dependent intracerebral theta activity within rostral subdivisions of the ACC. The posterior versus anterior index and theta current density within the rostral ACC were significantly correlated (r = -.52), and both displayed high retest stability across 5 hr and were associated with traits from the aE spectrum. These neurophysiological correlates of aE and their possible functional significance are discussed.

  8. Active power control of solar PV generation for large interconnection frequency regulation and oscillation damping

    DOE PAGES

    Liu, Yong; Zhu, Lin; Zhan, Lingwei; ...

    2015-06-23

    Because of zero greenhouse gas emission and decreased manufacture cost, solar photovoltaic (PV) generation is expected to account for a significant portion of future power grid generation portfolio. Because it is indirectly connected to the power grid via power electronic devices, solar PV generation system is fully decoupled from the power grid, which will influence the interconnected power grid dynamic characteristics as a result. In this study, the impact of solar PV penetration on large interconnected power system frequency response and inter-area oscillation is evaluated, taking the United States Eastern Interconnection (EI) as an example. Furthermore, based on the constructedmore » solar PV electrical control model with additional active power control loops, the potential contributions of solar PV generation to power system frequency regulation and oscillation damping are examined. The advantages of solar PV frequency support over that of wind generator are also discussed. Finally, simulation results demonstrate that solar PV generations can effectively work as ‘actuators’ in alleviating the negative impacts they bring about.« less

  9. Active power control of solar PV generation for large interconnection frequency regulation and oscillation damping

    SciTech Connect

    Liu, Yong; Zhu, Lin; Zhan, Lingwei; Gracia, Jose R.; King, Thomas Jr.; Liu, Yilu

    2015-06-23

    Because of zero greenhouse gas emission and decreased manufacture cost, solar photovoltaic (PV) generation is expected to account for a significant portion of future power grid generation portfolio. Because it is indirectly connected to the power grid via power electronic devices, solar PV generation system is fully decoupled from the power grid, which will influence the interconnected power grid dynamic characteristics as a result. In this study, the impact of solar PV penetration on large interconnected power system frequency response and inter-area oscillation is evaluated, taking the United States Eastern Interconnection (EI) as an example. Furthermore, based on the constructed solar PV electrical control model with additional active power control loops, the potential contributions of solar PV generation to power system frequency regulation and oscillation damping are examined. The advantages of solar PV frequency support over that of wind generator are also discussed. Finally, simulation results demonstrate that solar PV generations can effectively work as ‘actuators’ in alleviating the negative impacts they bring about.

  10. Design and Activation of a LOX/GH Chemical Steam Generator

    NASA Technical Reports Server (NTRS)

    Saunders, G. P.; Mulkey, C. A.; Taylor, S. A.

    2009-01-01

    The purpose of this paper is to give a detailed description of the design and activation of the LOX/GH fueled chemical steam generator installed in Cell 2 of the E3 test facility at Stennis Space Center, MS (SSC). The steam generator uses a liquid oxygen oxidizer with gaseous hydrogen fuel. The combustion products are then quenched with water to create steam at pressures from 150 to 450 psig at temperatures from 350 to 750 deg F (from saturation to piping temperature limits).

  11. Novel gene-by-environment interactions: APOB and NPC1L1 variants affect the relationship between dietary and total plasma cholesterol[S

    PubMed Central

    Kim, Daniel S.; Burt, Amber A.; Ranchalis, Jane E.; Jarvik, Ella R.; Rosenthal, Elisabeth A.; Hatsukami, Thomas S.; Furlong, Clement E.; Jarvik, Gail P.

    2013-01-01

    Cardiovascular disease (CVD) is the leading cause of death in developed countries. Plasma cholesterol level is a key risk factor in CVD pathogenesis. Genetic and dietary variation both influence plasma cholesterol; however, little is known about dietary interactions with genetic variants influencing the absorption and transport of dietary cholesterol. We sought to determine whether gut expressed variants predicting plasma cholesterol differentially affected the relationship between dietary and plasma cholesterol levels in 1,128 subjects (772/356 in the discovery/replication cohorts, respectively). Four single nucleotide polymorphisms (SNPs) within three genes (APOB, CETP, and NPC1L1) were significantly associated with plasma cholesterol in the discovery cohort. These were subsequently evaluated for gene-by-environment (GxE) interactions with dietary cholesterol for the prediction of plasma cholesterol, with significant findings tested for replication. Novel GxE interactions were identified and replicated for two variants: rs1042034, an APOB Ser4338Asn missense SNP and rs2072183 (in males only), a synonymous NPC1L1 SNP in linkage disequilibrium with SNPs 5′ of NPC1L1. This study identifies the presence of novel GxE and gender interactions implying that differential gut absorption is the basis for the variant associations with plasma cholesterol. These GxE interactions may account for part of the “missing heritability” not accounted for by genetic associations. PMID:23482652

  12. Active Oxygen Generator by Silent Discharge and Oxidation Power in Formation of Oxide Thin Films

    NASA Astrophysics Data System (ADS)

    Tanaka, Masaaki; Kawagoe, Yasuyuki; Tsukazaki, Hisashi; Yamanishi, Kenichiro

    We have studied the low pressure silent discharge type active oxygen generator in terms of the application to the formation of oxide thin films. In this paper the oxidation power of active oxygen in the oxide thin film formation is compared with that of oxygen and ozone by forming silicon oxide thin films. It was confirmed that the oxidation power is in turn of active oxygen > ozone > oxygen from the experimental result of the number of x in SiOx thin film. Furthermore we applied active oxygen to the formation of the thin film high temperature super conductor and active oxygen was found to be effective to the formation of the thin film with high performance.

  13. Auditory evoked fields to vocalization during passive listening and active generation in adults who stutter.

    PubMed

    Beal, Deryk S; Cheyne, Douglas O; Gracco, Vincent L; Quraan, Maher A; Taylor, Margot J; De Nil, Luc F

    2010-10-01

    We used magnetoencephalography to investigate auditory evoked responses to speech vocalizations and non-speech tones in adults who do and do not stutter. Neuromagnetic field patterns were recorded as participants listened to a 1 kHz tone, playback of their own productions of the vowel /i/ and vowel-initial words, and actively generated the vowel /i/ and vowel-initial words. Activation of the auditory cortex at approximately 50 and 100 ms was observed during all tasks. A reduction in the peak amplitudes of the M50 and M100 components was observed during the active generation versus passive listening tasks dependent on the stimuli. Adults who stutter did not differ in the amount of speech-induced auditory suppression relative to fluent speakers. Adults who stutter had shorter M100 latencies for the actively generated speaking tasks in the right hemisphere relative to the left hemisphere but the fluent speakers showed similar latencies across hemispheres. During passive listening tasks, adults who stutter had longer M50 and M100 latencies than fluent speakers. The results suggest that there are timing, rather than amplitude, differences in auditory processing during speech in adults who stutter and are discussed in relation to hypotheses of auditory-motor integration breakdown in stuttering.

  14. Indomethacin augments lymphokine-activated killer cell generation by patients with malignant mesothelioma

    SciTech Connect

    Manning, L.S.; Bowman, R.V.; Davis, M.R.; Musk, A.W.; Robinson, B.W. )

    1989-10-01

    Human malignant mesothelioma (MM) cells are resistant to natural killer (NK) cell lysis but susceptible to lysis by lymphokine-activated killer (LAK) cells from control individuals. The present study was performed to determine the capacity of patients with MM (n = 22) and individuals occupationally exposed to asbestos (the major population at risk of developing this disease, n = 52) to generate LAK cells capable of effectively lysing human mesothelioma cells. Compared to controls (n = 20), both patient groups demonstrated significantly depressed LAK cell activity against mesothelioma tumor cell targets (55 +/- 3% lysis by controls vs 34 +/- 3% lysis by patients with MM, P less than 0.005; and 45 +/- 3% lysis by asbestos-exposed individuals, P less than 0.025). Addition of 10 micrograms/ml indomethacin during LAK cell generation restored normal LAK cell activity for patients with MM (52 +/- 6% lysis of cultured human MM cells, P = NS compared to controls), suggesting that the defective cytolytic cell function observed in some patients with MM is a result of prostaglandin-induced immunosuppression. The ability of indomethacin to restore suppressed LAK cell activity in patients with MM suggests that the concomitant use of this agent in ex vivo LAK cell generation and in patients undergoing interleukin/LAK cell therapy may be beneficial.

  15. Systems Biology of Coagulation Initiation: Kinetics of Thrombin Generation in Resting and Activated Human Blood

    PubMed Central

    Chatterjee, Manash S.; Denney, William S.; Jing, Huiyan; Diamond, Scott L.

    2010-01-01

    Blood function defines bleeding and clotting risks and dictates approaches for clinical intervention. Independent of adding exogenous tissue factor (TF), human blood treated in vitro with corn trypsin inhibitor (CTI, to block Factor XIIa) will generate thrombin after an initiation time (Ti) of 1 to 2 hours (depending on donor), while activation of platelets with the GPVI-activator convulxin reduces Ti to ∼20 minutes. Since current kinetic models fail to generate thrombin in the absence of added TF, we implemented a Platelet-Plasma ODE model accounting for: the Hockin-Mann protease reaction network, thrombin-dependent display of platelet phosphatidylserine, VIIa function on activated platelets, XIIa and XIa generation and function, competitive thrombin substrates (fluorogenic detector and fibrinogen), and thrombin consumption during fibrin polymerization. The kinetic model consisting of 76 ordinary differential equations (76 species, 57 reactions, 105 kinetic parameters) predicted the clotting of resting and convulxin-activated human blood as well as predicted Ti of human blood under 50 different initial conditions that titrated increasing levels of TF, Xa, Va, XIa, IXa, and VIIa. Experiments with combined anti-XI and anti-XII antibodies prevented thrombin production, demonstrating that a leak of XIIa past saturating amounts of CTI (and not “blood-borne TF” alone) was responsible for in vitro initiation without added TF. Clotting was not blocked by antibodies used individually against TF, VII/VIIa, P-selectin, GPIb, protein disulfide isomerase, cathepsin G, nor blocked by the ribosome inhibitor puromycin, the Clk1 kinase inhibitor Tg003, or inhibited VIIa (VIIai). This is the first model to predict the observed behavior of CTI-treated human blood, either resting or stimulated with platelet activators. CTI-treated human blood will clot in vitro due to the combined activity of XIIa and XIa, a process enhanced by platelet activators and which proceeds in the

  16. Direct sequencing and characterization of a clinical isolate of Epstein-Barr virus from nasopharyngeal carcinoma tissue by using next-generation sequencing technology.

    PubMed

    Liu, Pan; Fang, Xiaodong; Feng, Zizhen; Guo, Yun-Miao; Peng, Rou-Jun; Liu, Tengfei; Huang, Zhiyong; Feng, Yue; Sun, Xiaoqing; Xiong, Zhiqiang; Guo, Xiaosen; Pang, Sha-Sha; Wang, Bo; Lv, Xiaojuan; Feng, Fu-Tuo; Li, Da-Jiang; Chen, Li-Zhen; Feng, Qi-Sheng; Huang, Wen-Lin; Zeng, Mu-Sheng; Bei, Jin-Xin; Zhang, Yong; Zeng, Yi-Xin

    2011-11-01

    Epstein-Barr virus (EBV)-encoded molecules have been detected in the tumor tissues of several cancers, including nasopharyngeal carcinoma (NPC), suggesting that EBV plays an important role in tumorigenesis. However, the nature of EBV with respect to genome width in vivo and whether EBV undergoes clonal expansion in the tumor tissues are still poorly understood. In this study, next-generation sequencing (NGS) was used to sequence DNA extracted directly from the tumor tissue of a patient with NPC. Apart from the human sequences, a clinically isolated EBV genome 164.7 kb in size was successfully assembled and named GD2 (GenBank accession number HQ020558). Sequence and phylogenetic analyses showed that GD2 was closely related to GD1, a previously assembled variant derived from a patient with NPC. GD2 contains the most prevalent EBV variants reported in Cantonese patients with NPC, suggesting that it might be the prevalent strain in this population. Furthermore, GD2 could be grouped into a single subtype according to common classification criteria and contains only 6 heterozygous point mutations, suggesting the monoclonal expansion of GD2 in NPC. This study represents the first genome-wide analysis of a clinical isolate of EBV directly extracted from NPC tissue. Our study reveals that NGS allows the characterization of genome-wide variations of EBV in clinical tumors and provides evidence of monoclonal expansion of EBV in vivo. The pipeline could also be applied to the study of other pathogen-related malignancies. With additional NGS studies of NPC, it might be possible to uncover the potential causative EBV variant involved in NPC.

  17. Recovery of rhythmic activity in a central pattern generator: analysis of the role of neuromodulator and activity-dependent mechanisms

    PubMed Central

    Zhang, Yili; Golowasch, Jorge

    2015-01-01

    The pyloric network of decapods crustaceans can undergo dramatic rhythmic activity changes. Under normal conditions the network generates low frequency rhythmic activity that depends obligatorily on the presence of neuromodulatory input from the central nervous system. When this input is removed (decentralization) the rhythmic activity ceases. In the continued absence of this input, periodic activity resumes after a few hours in the form of episodic bursting across the entire network that later turns into stable rhythmic activity that is nearly indistinguishable from control (recovery). It has been proposed that an activity-dependent modification of ionic conductance levels in the pyloric pacemaker neuron drives the process of recovery of activity. Previous modeling attempts have captured some aspects of the temporal changes observed experimentally, but key features could not be reproduced. Here we examined a model in which slow activity-dependent regulation of ionic conductances and slower neuromodulator-dependent regulation of intracellular Ca2+ concentration reproduce all the temporal features of this recovery. Key aspects of these two regulatory mechanisms are their independence and their different kinetics. We also examined the role of variability (noise) in the activity-dependent regulation pathway and observe that it can help to reduce unrealistic constraints that were otherwise required on the neuromodulator-dependent pathway. We conclude that small variations in intracellular Ca2+ concentration, a Ca2+ uptake regulation mechanism that is directly targeted by neuromodulator-activated signaling pathways, and variability in the Ca2+ concentration sensing signaling pathway can account for the observed changes in neuronal activity. Our conclusions are all amenable to experimental analysis. PMID:21573963

  18. A flexible active and reactive power control strategy for a variable speed constant frequency generating system

    SciTech Connect

    Tang, Y.; Xu, L.

    1995-07-01

    Variable-speed constant-frequency generating systems are used in wind power, hydro power, aerospace, and naval power generations to enhance efficiency and reduce friction. In these applications, an attractive candidate is the slip power recovery system comprising of doubly excited induction machine or doubly excited brushless reluctance machine and PWM converters with a dc link. In this paper, a flexible active and reactive power control strategy is developed, such that the optimal torque-speed profile of the turbine can be followed and overall reactive power can be controlled, while the machine copper losses have been minimized. At the same time, harmonics injected into the power network has also been minimized. In this manner, the system can function as both a high-efficient power generator and a flexible reactive power compensator.

  19. Platelet factor 4 enhances generation of activated protein C in vitro and in vivo.

    PubMed

    Slungaard, Arne; Fernandez, Jose A; Griffin, John H; Key, Nigel S; Long, Janel R; Piegors, Donald J; Lentz, Steven R

    2003-07-01

    Platelet factor 4 (PF4), an abundant platelet alpha-granule protein, accelerates in vitro generation of activated protein C (APC) by soluble thrombin/thrombomodulin (TM) complexes up to 25-fold. To test the hypothesis that PF4 similarly stimulates endothelium-associated TM, we assessed the influence of human PF4 on thrombin-dependent APC generation by cultured endothelial monolayers. APC generated in the presence of 1 to 100 microg PF4 was up to 5-fold higher than baseline for human umbilical vein endothelial cells, 10-fold higher for microvascular endothelial cells, and unaltered for blood outgrowth endothelial cells. In an in vivo model, cynomolgus monkeys (n = 6, each serving as its own control) were infused with either PF4 (7.5 mg/kg) or vehicle buffer, then with human thrombin (1.0 microg/kg/min) for 10 minutes. Circulating APC levels (baseline 3 ng/mL) peaked at 10 minutes, when PF4-treated and vehicle-treated animals had APC levels of 67 +/- 5 ng/mL and 39 +/- 2 ng/mL, respectively (P <.001). The activated partial thromboplastin time (APTT; baseline, 28 seconds) increased maximally by 27 +/- 6 seconds in PF4-treated animals and by 9 +/- 1 seconds in control animals at 30 minutes (P <.001). PF4-dependent increases in circulating APC and APTT persisted more than 2-fold greater than that of controls from 10 through 120 minutes (P < or =.04). All APTT prolongations were essentially reversed by monoclonal antibody C3, which blocks APC activity. Thus, physiologically relevant concentrations of PF4 stimulate thrombin-dependent APC generation both in vitro by cultured endothelial cells and in vivo in a primate thrombin infusion model. These findings suggest that PF4 may play a previously unsuspected physiologic role in enhancing APC generation.

  20. Effects of Globally Waste Disturbing Activities on Gas Generation, Retention, and Release in Hanford Waste Tanks

    SciTech Connect

    Stewart, Charles W.; Fountain, Matthew S.; Huckaby, James L.; Mahoney, Lenna A.; Meyer, Perry A.; Wells, Beric E.

    2005-08-02

    Various operations are authorized in Hanford single- and double-shell tanks that disturb all or a large fraction of the waste. These globally waste-disturbing activities have the potential to release a large fraction of the retained flammable gas and to affect future gas generation, retention, and release behavior. This report presents analyses of the expected flammable gas release mechanisms and the potential release rates and volumes resulting from these activities. The background of the flammable gas safety issue at Hanford is summarized, as is the current understanding of gas generation, retention, and release phenomena. Considerations for gas monitoring and assessment of the potential for changes in tank classification and steady-state flammability are given.

  1. Generation of unipolar optical pulses in a Raman-active medium

    NASA Astrophysics Data System (ADS)

    Arkhipov, R. M.; Arkhipov, M. V.; Belov, P. A.; Tolmachev, Yu A.; Babushkin, I.

    2016-04-01

    Response of a Raman-active media (RAM) to the excitation by a series of ultrashort (few-cycle) optical pulses propagating at a superluminal velocity is studied theoretically. It is shown that under certain conditions rectangular unipolar pulses (video-pulses) can be generated as the RAM response. The duration, shape and amplitude of these video-pulses can be widely tuned by modifying the pump pulse parameters.

  2. Isolation and Analysis of Novel Electrochemically Active Bacteria for Enhanced Power Generation in Microbial Fuel Cells

    DTIC Science & Technology

    2009-03-07

    ISOLATION AND ANALYSIS OF NOVEL ELECTROCHEMICALLY ACTIVE BACTERIA FOR ENHANCED POWER GENERATION IN MICROBIAL FUEL CELLS B.E. Logan, J.M. Regan...new exoelectrogenic bacteria during this project. We isolated Rhodopseudomonas palustris DX-1, and demonstrated for the first time that a pure culture... isolated Ochrobactrum anthropi YZ-1, which had the remarkable characteristic that it was unable to respire using hydrous Fe(lll) oxide but produced

  3. Time-Elastic Generative Model for Acceleration Time Series in Human Activity Recognition

    PubMed Central

    Munoz-Organero, Mario; Ruiz-Blazquez, Ramona

    2017-01-01

    Body-worn sensors in general and accelerometers in particular have been widely used in order to detect human movements and activities. The execution of each type of movement by each particular individual generates sequences of time series of sensed data from which specific movement related patterns can be assessed. Several machine learning algorithms have been used over windowed segments of sensed data in order to detect such patterns in activity recognition based on intermediate features (either hand-crafted or automatically learned from data). The underlying assumption is that the computed features will capture statistical differences that can properly classify different movements and activities after a training phase based on sensed data. In order to achieve high accuracy and recall rates (and guarantee the generalization of the system to new users), the training data have to contain enough information to characterize all possible ways of executing the activity or movement to be detected. This could imply large amounts of data and a complex and time-consuming training phase, which has been shown to be even more relevant when automatically learning the optimal features to be used. In this paper, we present a novel generative model that is able to generate sequences of time series for characterizing a particular movement based on the time elasticity properties of the sensed data. The model is used to train a stack of auto-encoders in order to learn the particular features able to detect human movements. The results of movement detection using a newly generated database with information on five users performing six different movements are presented. The generalization of results using an existing database is also presented in the paper. The results show that the proposed mechanism is able to obtain acceptable recognition rates (F = 0.77) even in the case of using different people executing a different sequence of movements and using different hardware. PMID

  4. Dissipative soliton generation in an active ring resonator based on magnonic quasicrystal with Fibonacci type structure

    NASA Astrophysics Data System (ADS)

    Grishin, S. V.; Beginin, E. N.; Sharaevskii, Yu. P.; Nikitov, S. A.

    2013-07-01

    This study reports on the experimental investigations of a magnetostatic surface wave (MSSW) propagation in a magnonic quasicrystal (MQC) with Fibonacci type structure. It is shown that such structure has a greater number of band gaps and narrower pass bands located between them than a periodic structure. These features of the MQC and three-wave decay of the MSSW are used in a MQC active ring resonator for the eigenmode selection and dissipative soliton self-generation.

  5. Generation of structurally novel short carotenoids and study of their biological activity.

    PubMed

    Kim, Se H; Kim, Moon S; Lee, Bun Y; Lee, Pyung C

    2016-02-23

    Recent research interest in phytochemicals has consistently driven the efforts in the metabolic engineering field toward microbial production of various carotenoids. In spite of systematic studies, the possibility of using C30 carotenoids as biologically functional compounds has not been explored thus far. Here, we generated 13 novel structures of C30 carotenoids and one C35 carotenoid, including acyclic, monocyclic, and bicyclic structures, through directed evolution and combinatorial biosynthesis, in Escherichia coli. Measurement of radical scavenging activity of various C30 carotenoid structures revealed that acyclic C30 carotenoids showed higher radical scavenging activity than did DL-α-tocopherol. We could assume high potential biological activity of the novel structures of C30 carotenoids as well, based on the neuronal differentiation activity observed for the monocyclic C30 carotenoid 4,4'-diapotorulene on rat bone marrow mesenchymal stem cells. Our results demonstrate that a series of structurally novel carotenoids possessing biologically beneficial properties can be synthesized in E. coli.

  6. Planning activity for internally generated reward goals in monkey amygdala neurons.

    PubMed

    Hernádi, István; Grabenhorst, Fabian; Schultz, Wolfram

    2015-03-01

    The best rewards are often distant and can only be achieved by planning and decision-making over several steps. We designed a multi-step choice task in which monkeys followed internal plans to save rewards toward self-defined goals. During this self-controlled behavior, amygdala neurons showed future-oriented activity that reflected the animal's plan to obtain specific rewards several trials ahead. This prospective activity encoded crucial components of the animal's plan, including value and length of the planned choice sequence. It began on initial trials when a plan would be formed, reappeared step by step until reward receipt, and readily updated with a new sequence. It predicted performance, including errors, and typically disappeared during instructed behavior. Such prospective activity could underlie the formation and pursuit of internal plans characteristic of goal-directed behavior. The existence of neuronal planning activity in the amygdala suggests that this structure is important in guiding behavior toward internally generated, distant goals.

  7. Coarse-grained simulations of an active filament propelled by a self-generated solute gradient

    NASA Astrophysics Data System (ADS)

    Sarkar, Debarati; Thakur, Snigdha

    2016-03-01

    A self-propelling semiflexible filament exhibits a variety of dynamical states depending on the flexibility and activity of the filament. Here we investigate the dynamics of such an active filament using a bead-spring model with the explicit hydrodynamic interactions. The activity in the filament is incorporated by inserting chemically active dimers at regular intervals along the chain. The chemical reactions at the catalytic bead of the dimer produces a self-generated concentration gradient and gives sufficient fuel to exhibit self-propulsion for the filament. Depending upon the rigidity and the configuration, the polymeric filament exhibits three distinct types of spontaneous motion, namely, rotational, snaking, and translational motion. The self-propulsion velocity of the filament for various rigidity and sizes has been calculated, and the factors affecting the propulsion are identified.

  8. Active fiber polymer cladding temperature measurement under conditions of laser generation and amplification

    NASA Astrophysics Data System (ADS)

    Sypin, V. E.; Prusakov, K. Y.; Ryabushkin, O. A.

    2016-04-01

    Polymer cladding temperature of active fiber in lasing regime is important parameter as it allows determination of fiber core temperature that in turn effects laser generation and amplification efficiency. Besides polymer cladding has much lower temperature damage threshold comparing to fused silica. For example, 200 degrees Kelvin overheating of the polymer cladding can result in fiber degradation. In present paper we introduce novel and simple method for precise temperature measurement of active fibers cladding under conditions of laser generation and amplification. Dependence of longitudinal temperature distribution along active fibers on optical pump power can be determined. This method employs measurement of temperature dependent electrical resistance of the metal wire being in thermal contact with fiber polymer cladding. The wire is reeled on the active fiber segment. Under lasing or amplification conditions the polymer cladding of the active fiber is heated together with coiled metal wire resulting in its electrical resistance change. By measuring resistance variation one can determine the temperature of the given fiber section.

  9. NO signalling decodes frequency of neuronal activity and generates synapse-specific plasticity in mouse cerebellum

    PubMed Central

    Namiki, Shigeyuki; Kakizawa, Sho; Hirose, Kenzo; Iino, Masamitsu

    2005-01-01

    Nitric oxide (NO) is an intercellular messenger regulating neuronal functions. To visualize NO signalling in the brain, we generated a novel fluorescent NO indicator, which consists of the heme-binding region (HBR) of soluble guanylyl cyclase and the green fluorescent protein. The indicator (HBR–GFP) was expressed in the Purkinje cells of the mouse cerebellum and we imaged NO signals in acute cerebellar slices upon parallel fibre (PF) activation with a train of burst stimulations (BS, each BS consisting of five pulses at 50 Hz). Our results showed that the intensity of synaptic NO signal decays steeply with the distance from the synaptic input near PF–Purkinje cell synapses and generates synapse-specific long-term potentiation (LTP). Furthermore, the NO release level has a bell-shaped dependence on the frequency of PF activity. At an optimal frequency (1 Hz), but not at a low frequency (0.25 Hz) of a train of 60 BS, NO release as well as LTP was induced. However, both NO release and LTP were significantly reduced at higher frequencies (2–4 Hz) of BS train due to cannabinoid receptor-mediated retrograde inhibition of NO generation at the PF terminals. These results suggest that synaptic NO signalling decodes the frequency of neuronal activity to mediate synaptic plasticity at the PF–Purkinje cell synapse. PMID:15919714

  10. [Central Pattern Generators: Mechanisms of the Activity and Their Role in the Control of "Automatic" Movements].

    PubMed

    Arshavsky, I; Deliagina, T G; Orlovsky, G N

    2015-01-01

    Central pattern generators (CPGs) are a set of interconnected neurons capable of generating a basic pattern of motor output underlying "automatic" movements (breathing, locomotion, chewing, swallowing, and so on) in the absence of afferent signals from the executive motor apparatus. They can be divided into the constitutive CPGs active throughout the entire lifetime (respiratory CPGs) and conditional CPGs controlling episodic movements (locomotion, chewing, swallowing, and others). Since a motor output of CPGs is determined by their internal organization, the activities of the conditional CPGs are initiated by simple commands coming from higher centers. We describe the structural and functional organization of the locomotor CPGs in the marine mollusk Clione limacina, lamprey, frog embryo, and laboratory mammals (cat, mouse, and rat), CPGs controlling the respiratory and swallowing movements in mammals, and CPGs controlling discharges of the electric organ in the gymnotiform fish. It is shown that in all these cases, the generation of rhythmic motor output is based both on the endogenous (pacemaker) activity of specific groups of interneurons and on interneural interactions. These two interrelated mechanisms complement each other, ensuring the high reliability of CPG functionality. We discuss how the experience obtained in studying CPGs can be used to understand mechanisms of more complex functions of the brain, including its cognitive functions.

  11. Thrombin generation by activated factor VII on platelet activated by different agonists. Extending the cell-based model of hemostasis

    PubMed Central

    Altman, Raul; Scazziota, Alejandra Silvia; Herrera, Maria de Lourdes; Gonzalez, Claudio

    2006-01-01

    Background Platelet activation is crucial in normal hemostasis. Using a clotting system free of external tissue factor, we investigated whether activated Factor VII in combination with platelet agonists increased thrombin generation (TG) in vitro. Methods and results TG was quantified by time parameters: lag time (LT) and time to peak (TTP), and by amount of TG: peak of TG (PTG) and area under thrombin formation curve after 35 minutes (AUC→35min) in plasma from 29 healthy volunteers using the calibrated automated thrombography (CAT) technique. TG parameters were measured at basal conditions and after platelet stimulation by sodium arachidonate (AA), ADP, and collagen (Col). In addition, the effects of recombinant activated FVII (rFVIIa) alone or combined with the other platelet agonists on TG parameters were investigated. We found that LT and TTP were significantly decreased (p < 0.05) and PTG and AUC→35min were significantly increased (p < 0.05) in platelet rich plasma activated with AA, ADP, Col, and rFVIIa compared to non-activated platelet rich plasma from normal subjects (p = 0.01). Furthermore platelet rich plasma activated by the combined effects of rFVIIa plus AA, ADP or Col had significantly reduced LT and TTP and increased AUC→35min (but not PTG) when compared to platelet rich plasma activated with agonists in the absence of rFVIIa. Conclusion Platelets activated by AA, ADP, Col or rFVIIa triggered TG. This effect was increased by combining rFVIIa with other agonists. Our intrinsic coagulation system produced a burst in TG independent of external tissue factor activity an apparent hemostatic effect with little thrombotic capacity. Thus we suggest a modification in the cell-based model of hemostasis. PMID:16630353

  12. Physical methods for generating and decoding neural activity in Hirudo verbana

    NASA Astrophysics Data System (ADS)

    Migliori, Benjamin John

    The interface between living nervous systems and hardware is an excellent proving ground for precision experimental methods and information classification systems. Nervous systems are complex (104 -- 10 15(!) connections), fragile, and highly active in intricate, constantly evolving patterns. However, despite the conveniently electrical nature of neural transmission, the interface between nervous systems and hardware poses significant experimental difficulties. As the desire for direct interfaces with neural signals continues to expand, the need for methods of generating and measuring neural activity with high spatiotemporal precision has become increasingly critical. In this thesis, I describe advances I have made in the ability to modify, generate, measure, and understand neural signals both in- and ex-vivo. I focus on methods developed for transmitting and extracting signals in the intact nervous system of Hirudo verbana (the medicinal leech), an animal with a minimally complex nervous system (10000 neurons distributed in packets along a nerve cord) that exhibits a diverse array of behaviors. To introduce artificial activity patterns, I developed a photothermal activation system in which a highly focused laser is used to irradiate carbon microparticles in contact with target neurons. The resulting local temperature increase generates an electrical current that forces the target neuron to fire neural signals, thereby providing a unique neural input mechanism. These neural signals can potentially be used to alter behavioral choice or generate specific behavioral output, and can be used endogenously in many animal models. I also describe new tools developed to expand the application of this method. In complement to this input system, I describe a new method of analyzing neural output signals involved in long-range coordination of behaviors. Leech behavioral signals are propagated between neural packets as electrical pulses in the nerve connective, a bundle of

  13. Helium generated cold plasma finely regulates activation of human fibroblast-like primary cells.

    PubMed

    Brun, Paola; Pathak, Surajit; Castagliuolo, Ignazio; Palù, Giorgio; Brun, Paola; Zuin, Matteo; Cavazzana, Roberto; Martines, Emilio

    2014-01-01

    Non-thermal atmospheric pressure plasmas are being developed for a wide range of health care applications, including wound healing. However in order to exploit the potential of plasma for clinical applications, the understanding of the mechanisms involved in plasma-induced activation of fibroblasts, the cells active in the healing process, is mandatory. In this study, the role of helium generated plasma in the tissue repairing process was investigated in cultured human fibroblast-like primary cells, and specifically in hepatic stellate cells and intestinal subepithelial myofibroblasts. Five minutes after treatment, plasma induced formation of reactive oxygen species (ROS) in cultured cells, as assessed by flow cytometric analysis of fluorescence-activated 2',7'-dichlorofluorescein diacetate probe. Plasma-induced intracellular ROS were characterized by lower concentrations and shorter half-lives with respect to hydrogen peroxide-induced ROS. Moreover ROS generated by plasma treatment increased the expression of peroxisome proliferator activated receptor (PPAR)-γ, nuclear receptor that modulates the inflammatory responses. Plasma exposure promoted wound healing in an in vitro model and induced fibroblast migration and proliferation, as demonstrated, respectively, by trans-well assay and partitioning between daughter cells of carboxyfluorescein diacetate succinimidyl ester fluorescent dye. Plasma-induced fibroblast migration and proliferation were found to be ROS-dependent as cellular incubation with antioxidant agents (e.g. N-acetyl L-cysteine) cancelled the biological effects. This study provides evidence that helium generated plasma promotes proliferation and migration in liver and intestinal fibroblast-like primary cells mainly by increasing intracellular ROS levels. Since plasma-evoked ROS are time-restricted and elicit the PPAR-γ anti-inflammatory molecular pathway, this strategy ensures precise regulation of human fibroblast activation and can be considered a

  14. Helium Generated Cold Plasma Finely Regulates Activation of Human Fibroblast-Like Primary Cells

    PubMed Central

    Brun, Paola; Pathak, Surajit; Castagliuolo, Ignazio; Palù, Giorgio; Brun, Paola; Zuin, Matteo; Cavazzana, Roberto; Martines, Emilio

    2014-01-01

    Non-thermal atmospheric pressure plasmas are being developed for a wide range of health care applications, including wound healing. However in order to exploit the potential of plasma for clinical applications, the understanding of the mechanisms involved in plasma-induced activation of fibroblasts, the cells active in the healing process, is mandatory. In this study, the role of helium generated plasma in the tissue repairing process was investigated in cultured human fibroblast-like primary cells, and specifically in hepatic stellate cells and intestinal subepithelial myofibroblasts. Five minutes after treatment, plasma induced formation of reactive oxygen species (ROS) in cultured cells, as assessed by flow cytometric analysis of fluorescence-activated 2′,7′-dichlorofluorescein diacetate probe. Plasma-induced intracellular ROS were characterized by lower concentrations and shorter half-lives with respect to hydrogen peroxide-induced ROS. Moreover ROS generated by plasma treatment increased the expression of peroxisome proliferator activated receptor (PPAR)-γ, nuclear receptor that modulates the inflammatory responses. Plasma exposure promoted wound healing in an in vitro model and induced fibroblast migration and proliferation, as demonstrated, respectively, by trans-well assay and partitioning between daughter cells of carboxyfluorescein diacetate succinimidyl ester fluorescent dye. Plasma-induced fibroblast migration and proliferation were found to be ROS-dependent as cellular incubation with antioxidant agents (e.g. N-acetyl L-cysteine) cancelled the biological effects. This study provides evidence that helium generated plasma promotes proliferation and migration in liver and intestinal fibroblast-like primary cells mainly by increasing intracellular ROS levels. Since plasma-evoked ROS are time-restricted and elicit the PPAR-γ anti-inflammatory molecular pathway, this strategy ensures precise regulation of human fibroblast activation and can be

  15. Leukotriene E4 activates peroxisome proliferator-activated receptor gamma and induces prostaglandin D2 generation by human mast cells.

    PubMed

    Paruchuri, Sailaja; Jiang, Yongfeng; Feng, Chunli; Francis, Sanjeev A; Plutzky, Jorge; Boyce, Joshua A

    2008-06-13

    Cysteinyl leukotrienes (cys-LTs) are potent inflammatory lipid mediators, of which leukotriene (LT) E(4) is the most stable and abundant in vivo. Although only a weak agonist of established G protein-coupled receptors (GPCRs) for cys-LTs, LTE(4) potentiates airway hyper-responsiveness (AHR) by a cyclooxygenase (COX)-dependent mechanism and induces bronchial eosinophilia. We now report that LTE(4) activates human mast cells (MCs) by a pathway involving cooperation between an MK571-sensitive GPCR and peroxisome proliferator-activated receptor (PPAR)gamma, a nuclear receptor for dietary lipids. Although LTD(4) is more potent than LTE(4) for inducing calcium flux by the human MC sarcoma line LAD2, LTE(4) is more potent for inducing proliferation and chemokine generation, and is at least as potent for upregulating COX-2 expression and causing prostaglandin D(2) (PGD(2)) generation. LTE(4) caused phosphorylation of extracellular signal-regulated kinase (ERK), p90RSK, and cyclic AMP-regulated-binding protein (CREB). ERK activation in response to LTE(4), but not to LTD(4), was resistant to inhibitors of phosphoinositol 3-kinase. LTE(4)-mediated COX-2 induction, PGD(2) generation, and ERK phosphorylation were all sensitive to interference by the PPARgamma antagonist GW9662 and to targeted knockdown of PPARgamma. Although LTE(4)-mediated PGD(2) production was also sensitive to MK571, an antagonist for the type 1 receptor for cys-LTs (CysLT(1)R), it was resistant to knockdown of this receptor. This LTE(4)-selective receptor-mediated pathway may explain the unique physiologic responses of human airways to LTE(4) in vivo.

  16. Iso standardization of theoretical activity evaluation method for low and intermediate level activated waste generated at nuclear power plants

    SciTech Connect

    Makoto Kashiwagi; Garamszeghy, Mike; Lantes, Bertrand; Bonne, Sebastien; Pillette-Cousin, Lucien; Leganes, Jose Luis; Volmert, Ben; James, David W.

    2013-07-01

    Disposal of low-and intermediate-level activated waste generated at nuclear power plants is being planned or carried out in many countries. The radioactivity concentrations and/or total quantities of long-lived, difficult-to-measure nuclides (DTM nuclides), such as C-14, Ni-63, Nb-94, α emitting nuclides etc., are often restricted by the safety case for a final repository as determined by each country's safety regulations, and these concentrations or amounts are required to be known and declared. With respect to waste contaminated by contact with process water, the Scaling Factor method (SF method), which is empirically based on sampling and analysis data, has been applied as an important method for determining concentrations of DTM nuclides. This method was standardized by the International Organization for Standardization (ISO) and published in 2007 as ISO21238 'Scaling factor method to determine the radioactivity of low and intermediate-level radioactive waste packages generated at nuclear power plants' [1]. However, for activated metal waste with comparatively high concentrations of radioactivity, such as may be found in reactor control rods and internal structures, direct sampling and radiochemical analysis methods to evaluate the DTM nuclides are limited by access to the material and potentially high personnel radiation exposure. In this case, theoretical calculation methods in combination with empirical methods based on remote radiation surveys need to be used to best advantage for determining the disposal inventory of DTM nuclides while minimizing exposure to radiation workers. Pursuant to this objective a standard for the theoretical evaluation of the radioactivity concentration of DTM nuclides in activated waste, is in process through ISO TC85/SC5 (ISO Technical Committee 85: Nuclear energy, nuclear technologies, and radiological protection; Subcommittee 5: Nuclear fuel cycle). The project team for this ISO standard was formed in 2011 and is composed of

  17. Motor cortex activation in Parkinson's disease: dissociation of electrocortical and peripheral measures of response generation.

    PubMed

    Praamstra, P; Plat, E M; Meyer, A S; Horstink, M W

    1999-09-01

    This study investigated characteristics of motor cortex activation and response generation in Parkinson's disease with measures of electrocortical activity (lateralized readiness potential [LRP]), electromyographic activity (EMG), and isometric force in a noise-compatibility task. When presented with stimuli consisting of incompatible target and distractor elements asking for responses of opposite hands, patients were less able than control subjects to suppress activation of the motor cortex controlling the wrong response hand. This was manifested in the pattern of reaction times and in an incorrect lateralization of the LRP. Onset latency and rise time of the LRP did not differ between patients and control subjects, but EMG and response force developed more slowly in patients. Moreover, in patients but not in control subjects, the rate of development of EMG and response force decreased as reaction time increased. We hypothesize that this dissociation between electrocortical activity and peripheral measures in Parkinson's disease is the result of changes in motor cortex function that alter the relation between signal-related and movement-related neural activity in the motor cortex. In the LRP, this altered balance may obscure an abnormal development of movement-related neural activity.

  18. Fish Swimming: Patternsin the Mechanical Energy Generation, Transmission and Dissipation from Muscle Activation to Body Movement

    NASA Astrophysics Data System (ADS)

    Zhang, W.; Yu, Y. L.; Tong, B. G.

    2011-09-01

    The power consumption of the undulatory fish swimming is produced by active muscles. The mechanical energy generated by stimulated muscles is dissipated partly by the passive tissues of fish while it is being transmitted to the fluid medium. Furthermore, the effective energy, propelling fish movement, is a part of that delivered by the fish body. The process depends on the interactions of the active muscles, the passive tissues, and the water surrounding the fish body. In the previous works, the body-fluid interactions have been investigated widely, but it is rarely considered how the mechanical energy generates, transmits and dissipates in fish swimming. This paper addresses the regular patterns of energy transfer process from muscle activation to body movement for a cruising lamprey (LAMPREY), a kind of anguilliform swimmer. It is necessary to propose a global modelling of the kinematic chain, which is composed of active muscle force-moment model, fish-body dynamic model and hydrodynamic model in order. The present results show that there are traveling energy waves along the fish body from anterior to posterior, accompanied with energy storing and dissipating due to the viscoelastic property of internal tissues. This study is a preliminary research on the framework of kinematic chain coordination performance in fish swimming.

  19. Reticular activating system of a central pattern generator: premovement electrical potentials.

    PubMed

    Tapia, Jesus A; Trejo, Argelia; Linares, Pablo; Alva, J Manuel; Kristeva, Rumyana; Manjarrez, Elias

    2013-10-01

    For the first time, here we characterize a bulbar reticular activating system (RAS) of neurons in decerebrate, deafferented and decerebellated cats producing a premovement electrical potential that we named obex slow potential (OSP). The OSP occurs about 0.8 ± 0.4 sec prior to the onset of a fictive-scratching-episode. Here, we describe two classes of bulbar neurons, off-on, which are silent but exhibit a 80 ± 56 Hz firing discharge at the beginning of (and during) the OSP, and on-off interneurons, with a 27 ± 14 Hz firing activity that stops at the beginning of (and during) the OSP. We suggest that these OSP-associated neurons belong to a descending RAS, which contributes to the activation of the spinal central pattern generators.

  20. Tailored motivational message generation: A model and practical framework for real-time physical activity coaching.

    PubMed

    Op den Akker, Harm; Cabrita, Miriam; Op den Akker, Rieks; Jones, Valerie M; Hermens, Hermie J

    2015-06-01

    This paper presents a comprehensive and practical framework for automatic generation of real-time tailored messages in behavior change applications. Basic aspects of motivational messages are time, intention, content and presentation. Tailoring of messages to the individual user may involve all aspects of communication. A linear modular system is presented for generating such messages. It is explained how properties of user and context are taken into account in each of the modules of the system and how they affect the linguistic presentation of the generated messages. The model of motivational messages presented is based on an analysis of existing literature as well as the analysis of a corpus of motivational messages used in previous studies. The model extends existing 'ontology-based' approaches to message generation for real-time coaching systems found in the literature. Practical examples are given on how simple tailoring rules can be implemented throughout the various stages of the framework. Such examples can guide further research by clarifying what it means to use e.g. user targeting to tailor a message. As primary example we look at the issue of promoting daily physical activity. Future work is pointed out in applying the present model and framework, defining efficient ways of evaluating individual tailoring components, and improving effectiveness through the creation of accurate and complete user- and context models.

  1. Genetic activation of BK currents in vivo generates bidirectional effects on neuronal excitability

    PubMed Central

    Montgomery, Jenna R.; Meredith, Andrea L.

    2012-01-01

    Large-conductance calcium-activated potassium channels (BK) are potent negative regulators of excitability in neurons and muscle, and increasing BK current is a novel therapeutic strategy for neuro- and cardioprotection, disorders of smooth muscle hyperactivity, and several psychiatric diseases. However, in some neurons, enhanced BK current is linked with seizures and paradoxical increases in excitability, potentially complicating the clinical use of agonists. The mechanisms that switch BK influence from inhibitory to excitatory are not well defined. Here we investigate this dichotomy using a gain-of-function subunit (BKR207Q) to enhance BK currents. Heterologous expression of BKR207Q generated currents that activated at physiologically relevant voltages in lower intracellular Ca2+, activated faster, and deactivated slower than wild-type currents. We then used BKR207Q expression to broadly augment endogenous BK currents in vivo, generating a transgenic mouse from a circadian clock-controlled Period1 gene fragment (Tg-BKR207Q). The specific impact on excitability was assessed in neurons of the suprachiasmatic nucleus (SCN) in the hypothalamus, a cell type where BK currents regulate spontaneous firing under distinct day and night conditions that are defined by different complements of ionic currents. In the SCN, Tg-BKR207Q expression converted the endogenous BK current to fast-activating, while maintaining similar current-voltage properties between day and night. Alteration of BK currents in Tg-BKR207Q SCN neurons increased firing at night but decreased firing during the day, demonstrating that BK currents generate bidirectional effects on neuronal firing under distinct conditions. PMID:23112153

  2. p53 activation contributes to patulin-induced nephrotoxicity via modulation of reactive oxygen species generation

    PubMed Central

    Jin, Huan; Yin, Shutao; Song, Xinhua; Zhang, Enxiang; Fan, Lihong; Hu, Hongbo

    2016-01-01

    Patulin is a major mycotoxin found in fungal contaminated fruits and their derivative products. Previous studies showed that patulin was able to induce increase of reactive oxygen species (ROS) generation and oxidative stress was suggested to play a pivotal role in patulin-induced multiple toxic signaling. The objective of the present study was to investigate the functional role of p53 in patulin-induced oxidative stress. Our study demonstrated that higher levels of ROS generation and DNA damage were induced in wild-type p53 cell lines than that found in either knockdown or knockout p53 cell lines in response to patulin exposure, suggesting p53 activation contributed to patulin-induced ROS generation. Mechanistically, we revealed that the pro-oxidant role of p53 in response to patulin was attributed to its ability to suppress catalase activity through up-regulation of PIG3. Moreover, these in vitro findings were further validated in the p53 wild-type/knockout mouse model. To the best of our knowledge, this is the first report addressing the functional role of p53 in patulin-induced oxidative stress. The findings of the present study provided novel insights into understanding mechanisms behind oxidative stress in response to patulin exposure. PMID:27071452

  3. Reducing GBA2 Activity Ameliorates Neuropathology in Niemann-Pick Type C Mice

    PubMed Central

    Ottenhoff, Roelof; van Roomen, Cindy P. A. A.; Herrera Moro, Daniela; Claessen, Nike; Vinueza Veloz, María Fernanda; Zhou, Kuikui; Lin, Zhanmin; Mirzaian, Mina; Boot, Rolf G.; De Zeeuw, Chris I.; Overkleeft, Herman S.; Yildiz, Yildiz; Aerts, Johannes M. F. G.

    2015-01-01

    The enzyme glucocerebrosidase (GBA) hydrolyses glucosylceramide (GlcCer) in lysosomes. Markedly reduced GBA activity is associated with severe manifestations of Gaucher disease including neurological involvement. Mutations in the GBA gene have recently also been identified as major genetic risk factor for Parkinsonism. Disturbed metabolism of GlcCer may therefore play a role in neuropathology. Besides lysosomal GBA, cells also contain a non-lysosomal glucosylceramidase (GBA2). Given that the two β-glucosidases share substrates, we speculated that over-activity of GBA2 during severe GBA impairment might influence neuropathology. This hypothesis was studied in Niemann-Pick type C (Npc1-/-) mice showing secondary deficiency in GBA in various tissues. Here we report that GBA2 activity is indeed increased in the brain of Npc1-/- mice. We found that GBA2 is particularly abundant in Purkinje cells (PCs), one of the most affected neuronal populations in NPC disease. Inhibiting GBA2 in Npc1-/- mice with a brain-permeable low nanomolar inhibitor significantly improved motor coordination and extended lifespan in the absence of correction in cholesterol and ganglioside abnormalities. This trend was recapitulated, although not to full extent, by introducing a genetic loss of GBA2 in Npc1-/- mice. Our findings point to GBA2 activity as therapeutic target in NPC. PMID:26275242

  4. Feijoa sellowiana Berg fruit juice: anti-inflammatory effect and activity on superoxide anion generation.

    PubMed

    Monforte, Maria T; Fimiani, Vincenzo; Lanuzza, Francesco; Naccari, Clara; Restuccia, Salvatore; Galati, Enza M

    2014-04-01

    Feijoa sellowiana Berg var. coolidge fruit juice was studied in vivo for the anti-inflammatory activity by carrageenin-induced paw edema test and in vitro for the effects on superoxide anion release from neutrophils in human whole blood. The fruit juice was analyzed by the high-performance liquid chromatography method, and quercetin, ellagic acid, catechin, rutin, eriodictyol, gallic acid, pyrocatechol, syringic acid, and eriocitrin were identified. The results showed a significant anti-inflammatory activity of F. sellowiana fruit juice, sustained also by an effective antioxidant activity observed in preliminary studies on 1,1-diphenyl-2-picrylhydrazyl (DPPH) test. In particular, the anti-inflammatory activity edema inhibition is significant since the first hour (44.11%) and persists until the fifth hour (44.12%) of the treatment. The effect on superoxide anion release was studied in human whole blood, in the presence of activators affecting neutrophils by different mechanisms. The juice showed an inhibiting response on neutrophils basal activity in all experimental conditions. In stimulated neutrophils, the higher inhibition of superoxide anion generation was observed at concentration of 10(-4) and 10(-2) mg/mL in whole blood stimulate with phorbol-myristate-13-acetate (PMA; 20% and 40%) and with N-formyl-methionyl-leucyl-phenylalanine (FMLP; 15% and 48%). The significant reduction of edema and the inhibition of O2(-) production, occurring mainly through interaction with protein-kinase C pathway, confirm the anti-inflammatory effect of F. sellowiana fruit juice.

  5. Analysis of crystal structure of Arabidopsis MPK6 and generation of its mutants with higher activity.

    PubMed

    Wang, Bo; Qin, Xinghua; Wu, Juan; Deng, Hongying; Li, Yuan; Yang, Hailian; Chen, Zhongzhou; Liu, Guoqin; Ren, Dongtao

    2016-05-10

    Mitogen-activated protein kinase (MAPK) cascades, which are the highly conserved signalling modules in eukaryotic organisms, have been shown to play important roles in regulating growth, development, and stress responses. The structures of various MAPKs from yeast and animal have been solved, and structure-based mutants were generated for their function analyses, however, the structures of plant MAPKs remain unsolved. Here, we report the crystal structure of Arabidopsis MPK6 at a 3.0 Å resolution. Although MPK6 is topologically similar to ERK2 and p38, the structures of the glycine-rich loop, MAPK insert, substrate binding sites, and L16 loop in MPK6 show notable differences from those of ERK2 and p38. Based on the structural comparison, we constructed MPK6 mutants and analyzed their kinase activity both in vitro and in planta. MPK6(F364L) and MPK6(F368L) mutants, in which Phe364 and Phe368 in the L16 loop were changed to Leu, respectively, acquired higher intrinsic kinase activity and retained the normal MAPKK activation property. The expression of MPK6 mutants with basal activity is sufficient to induce camalexin biosynthesis; however, to induce ethylene and leaf senescence, the expression of MPK6 mutants with higher activity is required. The results suggest that these mutants can be used to analyze the specific biological functions of MPK6.

  6. Active integration of glutamatergic input to the inferior olive generates bidirectional postsynaptic potentials

    PubMed Central

    Garden, Derek L. F.; Rinaldi, Arianna

    2016-01-01

    Key points We establish experimental preparations for optogenetic investigation of glutamatergic input to the inferior olive.Neurones in the principal olivary nucleus receive monosynaptic extra‐somatic glutamatergic input from the neocortex.Glutamatergic inputs to neurones in the inferior olive generate bidirectional postsynaptic potentials (PSPs), with a fast excitatory component followed by a slower inhibitory component.Small conductance calcium‐activated potassium (SK) channels are required for the slow inhibitory component of glutamatergic PSPs and oppose temporal summation of inputs at intervals ≤ 20 ms.Active integration of synaptic input within the inferior olive may play a central role in control of olivo‐cerebellar climbing fibre signals. Abstract The inferior olive plays a critical role in motor coordination and learning by integrating diverse afferent signals to generate climbing fibre inputs to the cerebellar cortex. While it is well established that climbing fibre signals are important for motor coordination, the mechanisms by which neurones in the inferior olive integrate synaptic inputs and the roles of particular ion channels are unclear. Here, we test the hypothesis that neurones in the inferior olive actively integrate glutamatergic synaptic inputs. We demonstrate that optogenetically activated long‐range synaptic inputs to the inferior olive, including projections from the motor cortex, generate rapid excitatory potentials followed by slower inhibitory potentials. Synaptic projections from the motor cortex preferentially target the principal olivary nucleus. We show that inhibitory and excitatory components of the bidirectional synaptic potentials are dependent upon AMPA (GluA) receptors, are GABAA independent, and originate from the same presynaptic axons. Consistent with models that predict active integration of synaptic inputs by inferior olive neurones, we find that the inhibitory component is reduced by blocking large conductance

  7. Active control of highly efficient third-harmonic generation in ultrathin nonlinear metasurfaces

    NASA Astrophysics Data System (ADS)

    Gong, Zibo; Li, Chong; Hu, Xiaoyong; Yang, Hong; Gong, Qihuang

    2016-10-01

    Active electric control of highly efficient third harmonic generation was realized in an ultrathin nonlinear metasurface by using a nanocomposite consisting of gold nanoparticles dispersed in polycrystalline strontium titanate as the electro-optic material. Owing to the nonlinearity enhancement associated with the slow light effect, quantum confinement effect, and field-reinforcement, a high conversion efficiency of 3 × 10-5 was obtained, which is two orders of magnitude larger than previously reported efficiencies at comparable pump intensities. A modulation of 12% in the intensity of the third harmonic generation and a 30-nm shift in the transparency window center were achieved by varying the applied voltage from -30 V to zero. Our results pave the way toward the realization of multi-functional integrated photonic devices and chips based on metasurfaces.

  8. Three-dimensional analysis of optical forces generated by an active tractor beam using radial polarization.

    PubMed

    Carretero, Luis; Acebal, Pablo; Blaya, Salvador

    2014-02-10

    We theoretically study the three-dimensional behavior of nanoparticles in an active optical conveyor. To do this, we solved the Langevin equation when the forces are generated by a focusing system at the near field. Analytical expressions for the optical forces generated by the optical conveyor were obtained by solving the Richards and Wolf vectorial diffraction integrals in an approximated form when a mask of two annular pupils is illuminated by a radially polarized Hermite-Gauss beam. Trajectories, in both the transverse plane and the longitudinal direction, are analyzed showing that the behavior of the optical conveyor can be optimized by conveniently choosing the configuration of the mask of the two annular pupils (inner and outer radius of the two rings) in order to trap and transport all particles at the focal plane.

  9. [Kinetic modeling of energy metabolism and generation of active forms of oxygen in hepatocyte mitochondria].

    PubMed

    Demin, O V; Gorianin, I I; Kholodenko, B N; Westerhoff, H V

    2001-01-01

    Direct nonenzymatic oxidation of semiquinone by oxygen is one of the main sources of superoxide radicals (O2.-) in mitochondria. By using all the known data on hepatocyte mitochondria, we have revealed the correlation between the rate of superoxide generation by the bc1 complex and the transmembrane potential (delta psi). If the main electrogenic stage of the Q cycle is suggested to be the electron transfer between the cytochrome b hemes, then the rate of superoxide generation sharply increases when delta psi grows from 150 mV to 180 mV. However, this interrelation is ambiguous. Indeed, the increase of the generation rate with the growth of the potential can occur faster when succinate dehydrogenase is inhibited by malonate than when external ADP is exhausted. When the potential is changed by adding phosphate or potassium (K+), the rate of O2.- production remains constant, although the comparison of the rate values at the same delta psi reveals the effect of phosphate or potassium. It turned out that the rate of O2.- generation is a function of delta mu H rather than any of its components. Phosphate and K+ have practically no influence on delta mu H, since the change in delta psi is compensated by delta pH. The rate of superoxide generation by the bc1 complex is a multiple function of the electron-transfer activity of enzymes, the processes determining the membrane potential (e.g., loading), and of the oxygen concentration. The kinetic model proposed in this work may serve a tool to understand how the superoxide production is regulated.

  10. The Second-Generation Exportin-1 Inhibitor KPT-8602 Demonstrates Potent Activity against Acute Lymphoblastic Leukemia.

    PubMed

    Vercruysse, Thomas; De Bie, Jolien; Neggers, Jasper E; Jacquemyn, Maarten; Vanstreels, Els; Schmid-Burgk, Jonathan L; Hornung, Veit; Baloglu, Erkan; Landesman, Yosef; Senapedis, William; Shacham, Sharon; Dagklis, Antonis; Cools, Jan; Daelemans, Dirk

    2016-10-25

    Purpose: Human exportin-1 (XPO1) is the key nuclear-cytoplasmic transport protein that exports different cargo proteins out of the nucleus. Inducing nuclear accumulation of these proteins by inhibiting XPO1 causes cancer cell death. First clinical validation of pharmacological inhibition of XPO1 was obtained with the Selective Inhibitor of Nuclear Export (SINE) compound selinexor (KPT-330) demonstrating activity in phase-II/IIb clinical trials when dosed 1 to 3 times weekly. The second-generation SINE compound KPT-8602 shows improved tolerability and can be dosed daily. Here, we investigate and validate the drug-target interaction of KPT-8602 and explore its activity against acute lymphoblastic leukemia (ALL).Experimental Design: We examined the effect of KPT-8602 on XPO1 function and XPO1-cargo as well as on a panel of leukemia cell lines. Mutant XPO1 leukemia cells were designed to validate KPT-8602's drug-target interaction. In vivo, anti-ALL activity was measured in a mouse ALL model and patient-derived ALL xenograft models.Results: KPT-8602 induced caspase-dependent apoptosis in a panel of leukemic cell lines in vitro Using CRISPR/Cas9 genome editing, we demonstrated the specificity of KPT-8602 for cysteine 528 in the cargo-binding groove of XPO1 and validated the drug target interaction. In vivo, KPT-8602 showed potent anti-leukemia activity in a mouse ALL model as well as in patient-derived T- and B-ALL xenograft models without affecting normal hematopoiesis.Conclusions: KPT-8602 is highly specific for XPO1 inhibition and demonstrates potent anti-leukemic activity supporting clinical application of the second-generation SINE compound for the treatment of ALL. Clin Cancer Res; 1-14. ©2016 AACR.

  11. Development of high flux thermal neutron generator for neutron activation analysis

    NASA Astrophysics Data System (ADS)

    Vainionpaa, Jaakko H.; Chen, Allan X.; Piestrup, Melvin A.; Gary, Charles K.; Jones, Glenn; Pantell, Richard H.

    2015-05-01

    The new model DD110MB neutron generator from Adelphi Technology produces thermal (<0.5 eV) neutron flux that is normally achieved in a nuclear reactor or larger accelerator based systems. Thermal neutron fluxes of 3-5 · 107 n/cm2/s are measured. This flux is achieved using four ion beams arranged concentrically around a target chamber containing a compact moderator with a central sample cylinder. Fast neutron yield of ∼2 · 1010 n/s is created at the titanium surface of the target chamber. The thickness and material of the moderator is selected to maximize the thermal neutron flux at the center. The 2.5 MeV neutrons are quickly thermalized to energies below 0.5 eV and concentrated at the sample cylinder. The maximum flux of thermal neutrons at the target is achieved when approximately half of the neutrons at the sample area are thermalized. In this paper we present simulation results used to characterize performance of the neutron generator. The neutron flux can be used for neutron activation analysis (NAA) prompt gamma neutron activation analysis (PGNAA) for determining the concentrations of elements in many materials. Another envisioned use of the generator is production of radioactive isotopes. DD110MB is small enough for modest-sized laboratories and universities. Compared to nuclear reactors the DD110MB produces comparable thermal flux but provides reduced administrative and safety requirements and it can be run in pulsed mode, which is beneficial in many neutron activation techniques.

  12. Motor Neurons Tune Premotor Activity in a Vertebrate Central Pattern Generator.

    PubMed

    Lawton, Kristy J; Perry, Wick M; Yamaguchi, Ayako; Zornik, Erik

    2017-03-22

    Central patterns generators (CPGs) are neural circuits that drive rhythmic motor output without sensory feedback. Vertebrate CPGs are generally believed to operate in a top-down manner in which premotor interneurons activate motor neurons that in turn drive muscles. In contrast, the frog (Xenopus laevis) vocal CPG contains a functionally unexplored neuronal projection from the motor nucleus to the premotor nucleus, indicating a recurrent pathway that may contribute to rhythm generation. In this study, we characterized the function of this bottom-up connection. The X. laevis vocal CPG produces a 50-60 Hz "fast trill" song used by males during courtship. We recorded "fictive vocalizations" in the in vitro CPG from the laryngeal nerve while simultaneously recording premotor activity at the population and single-cell level. We show that transecting the motor-to-premotor projection eliminated the characteristic firing rate of premotor neurons. Silencing motor neurons with the intracellular sodium channel blocker QX-314 also disrupted premotor rhythms, as did blockade of nicotinic synapses in the motor nucleus (the putative location of motor neuron-to-interneuron connections). Electrically stimulating the laryngeal nerve elicited primarily IPSPs in premotor neurons that could be blocked by a nicotinic receptor antagonist. Our results indicate that an inhibitory signal, activated by motor neurons, is required for proper CPG function. To our knowledge, these findings represent the first example of a CPG in which precise premotor rhythms are tuned by motor neuron activity.SIGNIFICANCE STATEMENT Central pattern generators (CPGs) are neural circuits that produce rhythmic behaviors. In vertebrates, motor neurons are not commonly known to contribute to CPG function, with the exception of a few spinal circuits where the functional significance of motor neuron feedback is still poorly understood. The frog hindbrain vocal circuit contains a previously unexplored connection from

  13. Characterization of sacral interneurons that mediate activation of locomotor pattern generators by sacrocaudal afferent input.

    PubMed

    Etlin, Alex; Finkel, Eran; Mor, Yoav; O'Donovan, Michael J; Anglister, Lili; Lev-Tov, Aharon

    2013-01-09

    Identification of the neural pathways involved in retraining the spinal central pattern generators (CPGs) by afferent input in the absence of descending supraspinal control is feasible in isolated rodent spinal cords where the locomotor CPGs are potently activated by sacrocaudal afferent (SCA) input. Here we study the involvement of sacral neurons projecting rostrally through the ventral funiculi (VF) in activation of the CPGs by sensory stimulation. Fluorescent labeling and immunostaining showed that VF neurons are innervated by primary afferents immunoreactive for vesicular glutamate transporters 1 and 2 and by intraspinal neurons. Calcium imaging revealed that 55% of the VF neurons were activated by SCA stimulation. The activity of VF neurons and the sacral and lumbar CPGs was abolished when non-NMDA receptors in the sacral segments were blocked by the antagonist CNQX. When sacral NMDA receptors were blocked by APV, the sacral CPGs were suppressed, VF neurons with nonrhythmic activity were recruited and a moderate-drive locomotor rhythm developed during SCA stimulation. In contrast, when the sacral CPGs were activated by SCA stimulation, rhythmic and nonrhythmic VF neurons were recruited and the locomotor rhythm was most powerful. The activity of 73 and 27% of the rhythmic VF neurons was in-phase with the ipsilateral and contralateral motor output, respectively. Collectively, our studies indicate that sacral VF neurons serve as a major link between SCA and the hindlimb CPGs and that the ability of SCA to induce stepping can be enhanced by the sacral CPGs. The nature of the ascending drive to lumbar CPGs, the identity of subpopulations of VF neurons, and their potential role in activating the locomotor rhythm are discussed.

  14. Transform-limited pulses generated by an actively Q-switched distributed fiber laser.

    PubMed

    Cuadrado-Laborde, C; Pérez-Millán, P; Andrés, M V; Díez, A; Cruz, J L; Barmenkov, Yu O

    2008-11-15

    A single-mode, transform-limited, actively Q-switched distributed-feedback fiber laser is presented, based on a new in-line acoustic pulse generator. Our technique permits a continuous adjustment of the repetition rate that modulates the Q factor of the cavity. Optical pulses of 800 mW peak power, 32 ns temporal width, and up to 20 kHz repetition rates were obtained. The measured linewidth demonstrates that these pulses are transform limited: 6 MHz for a train of pulses of 10 kHz repetition rate, 80 ns temporal width, and 60 mW peak power. Efficient excitation of spontaneous Brillouin scattering is demonstrated.

  15. Testing and Oxygen Assessment Results for a Next Generation Extravehicular Activity Portable Life Support System Fan

    NASA Technical Reports Server (NTRS)

    Paul, Heather L.; Jennings, Mallory A.; Rivera, Fatonia L.; Martin, Devin

    2011-01-01

    NASA is designing a next generation Extravehicular Activity (EVA) Portable Life Support System (PLSS) for use in future surface exploration endeavors. To meet the new requirements for ventilation flow at nominal and buddy modes, a fan has been developed and tested. This paper summarizes the results of the performance and life cycle testing efforts conducted at the NASA Johnson Space Center. Additionally, oxygen compatibility assessment results from an evaluation conducted at White Sands Test Facility (WSTF) are provided, and lessons learned and future recommendations are outlined.

  16. Automation of labelling of Lipiodol with high-activity generator-produced 188Re.

    PubMed

    Lepareur, Nicolas; Ardisson, Valérie; Noiret, Nicolas; Boucher, Eveline; Raoul, Jean-Luc; Clément, Bruno; Garin, Etienne

    2011-02-01

    This work describes optimisation of the kit formulation for labelling of Lipiodol with high-activity generator-produced rhenium-188. Radiochemical purity (RCP) was 92.52±2.3% and extraction yield was 98.56±1.2%. The synthesis has been automated with a TADDEO module (Comecer) giving a mean final yield of 52.68±9.6%, and reducing radiation burden to the radiochemist by 80%. Radiolabelled Lipiodol ((188)Re-SSS/Lipiodol) is stable for at least 7 days (RCP=91.07±0.9%).

  17. Active force generation in cross-linked filament bundles without motor proteins.

    PubMed

    Walcott, Sam; Sun, Sean X

    2010-11-01

    Cytoskeletal filaments often interact laterally through cross-linking proteins, contributing to passive cellular viscoelasticity and, perhaps surprisingly, active force generation. We present a theory, based on the formation and rupture of cross-linker bonds, that relates molecular properties of those interactions to the macroscale mechanics of filament bundles. Computing the force-velocity relation for such a bundle, we demonstrate significant contractile forces in the absence of molecular motors. This theory provides insight into cytokinesis, cytoskeletal mechanics, and stress-fiber contraction.

  18. Alliinase from Ensifer adhaerens and Its Use for Generation of Fungicidal Activity

    PubMed Central

    2011-01-01

    A bacterium Ensifer adhaerens FERM P-19486 with the ability of alliinase production was isolated from a soil sample. The enzyme was purified for characterization of its general properties and evaluation of its application in on-site production of allicin-dependent fungicidal activity. The bacterial alliinase was purified 300-fold from a cell-free extract, giving rise to a homogenous protein band on polyacrylamide gel electrophoresis. The bacterial alliinase (96 kDa) consisted of two identical subunits (48 kDa), and was most active at 60°C and at pH 8.0. The enzyme stoichiometrically converted (-)-alliin ((-)-S-allyl-L-cysteine sulfoxide) to form allicin, pyruvic acid, and ammonia more selectively than (+)-alliin, a naturally occurring substrate for plant alliinase ever known. The C-S lyase activity was also detected with this bacterial enzyme when S-alkyl-L-cysteine was used as a substrate, though such a lyase activity is absolutely absent in alliinase of plant origin. The enzyme generated a fungicidal activity against Saccharomyces cerevisiae in a time- and a dose-dependent fashion using alliin as a stable precursor. Alliinase of Ensifer adhaerens FERM P-19486 is the enzyme with a novel type of substrate specificity, and thus considered to be beneficial when used in combination with garlic enzyme with respect to absolute conversion of (±)-alliin to allicin. PMID:21906328

  19. Efficient peroxydisulfate activation process not relying on sulfate radical generation for water pollutant degradation.

    PubMed

    Zhang, Tao; Chen, Yin; Wang, Yuru; Le Roux, Julien; Yang, Yang; Croué, Jean-Philippe

    2014-05-20

    Peroxydisulfate (PDS) is an appealing oxidant for contaminated groundwater and toxic industrial wastewaters. Activation of PDS is necessary for application because of its low reactivity. Present activation processes always generate sulfate radicals as actual oxidants which unselectively oxidize organics and halide anions reducing oxidation capacity of PDS and producing toxic halogenated products. Here we report that copper oxide (CuO) can efficiently activate PDS under mild conditions without producing sulfate radicals. The PDS/CuO coupled process is most efficient at neutral pH for decomposing a model compound, 2,4-dichlorophenol (2,4-DCP). In a continuous-flow reaction with an empty-bed contact time of 0.55 min, over 90% of 2,4-DCP (initially 20 μM) and 90% of adsorbable organic chlorine (AOCl) can be removed at the PDS/2,4-DCP molar ratio of 1 and 4, respectively. Based on kinetic study and surface characterization, PDS is proposed to be first activated by CuO through outer-sphere interaction, the rate-limiting step, followed by a rapid reaction with 2,4-DCP present in the solution. In the presence of ubiquitous chloride ions in groundwater/industrial wastewater, the PDS/CuO oxidation shows significant advantages over sulfate radical oxidation by achieving much higher 2,4-DCP degradation capacity and avoiding the formation of highly chlorinated degradation products. This work provides a new way of PDS activation for contaminant removal.

  20. Optimizing for generalization in the decoding of internally generated activity in the hippocampus.

    PubMed

    van der Meer, Matthijs A A; Carey, Alyssa A; Tanaka, Youki

    2017-02-08

    The decoding of a sensory or motor variable from neural activity benefits from a known ground truth against which decoding performance can be compared. In contrast, the decoding of covert, cognitive neural activity, such as occurs in memory recall or planning, typically cannot be compared to a known ground truth. As a result, it is unclear how decoders of such internally generated activity should be configured in practice. We suggest that if the true code for covert activity is unknown, decoders should be optimized for generalization performance using cross-validation. Using ensemble recording data from hippocampal place cells, we show that this cross-validation approach results in different decoding error, different optimal decoding parameters, and different distributions of error across the decoded variable space. In addition, we show that a minor modification to the commonly used Bayesian decoding procedure, which enables the use of spike density functions, results in substantially lower decoding errors. These results have implications for the interpretation of covert neural activity, and suggest easy-to-implement changes to commonly used procedures across domains, with applications to hippocampal place cells in particular. © 2017 Wiley Periodicals, Inc.

  1. Generation of knockout rabbits using transcription activator-like effector nucleases.

    PubMed

    Wang, Yu; Fan, Nana; Song, Jun; Zhong, Juan; Guo, Xiaogang; Tian, Weihua; Zhang, Quanjun; Cui, Fenggong; Li, Li; Newsome, Philip N; Frampton, Jon; Esteban, Miguel A; Lai, Liangxue

    2014-01-01

    Zinc-finger nucleases and transcription activator-like effector nucleases are novel gene-editing platforms contributing to redefine the boundaries of modern biological research. They are composed of a non-specific cleavage domain and a tailor made DNA-binding module, which enables a broad range of genetic modifications by inducing efficient DNA double-strand breaks at desired loci. Among other remarkable uses, these nucleases have been employed to produce gene knockouts in mid-size and large animals, such as rabbits and pigs, respectively. This approach is cost effective, relatively quick, and can produce invaluable models for human disease studies, biotechnology or agricultural purposes. Here we describe a protocol for the efficient generation of knockout rabbits using transcription activator-like effector nucleases, and a perspective of the field.

  2. Experiments and modelling of active quasi-single helicity regime generation in a reversed field pinch

    NASA Astrophysics Data System (ADS)

    Frassinetti, L.; Brunsell, P. R.; Drake, J. R.

    2009-07-01

    The interaction of a static resonant magnetic perturbation (RMP) with a tearing mode (TM) is becoming a relevant topic in fusion plasma physics. RMPs can be generated by active coils and then used to affect the properties of TMs and of the corresponding magnetic islands. This paper shows how the feedback system of the EXTRAP T2R reversed field pinch (RFP) can produce a RMP that affects a rotating TM and stimulate the transition to the so-called quasi-single helicity (QSH) regime, a RFP plasma state characterized by a magnetic island surrounded by low magnetic chaos. The application of the RMP can increase the QSH probability up to 10% and enlarge the size of the corresponding island. Part of the experimental results are supported by a theoretical study that models the effect of the active coils on the magnetic island.

  3. Multicomponent, Mannich-type assembly process for generating novel, biologically-active 2-arylpiperidines and derivatives

    PubMed Central

    Hardy, Simon; Martin, Stephen F.

    2014-01-01

    A multicomponent, Mannich-type assembly process commencing with commercially available bromobenzaldehydes was sequenced with [3+2] dipolar cycloaddition reactions involving nitrones and azomethine ylides to generate collections of fused, bicyclic scaffolds based on the 2-arylpiperidine subunit. Use of the 4-pentenoyl group, which served both as an activator in the Mannich-type reaction and a readily-cleaved amine protecting group, allowed sub-libraries to be prepared through piperidine N-functionalization and cross-coupling of the aryl bromide. A number of these derivatives displayed biological activities that had not previously been associated with this substructure. Methods were also developed that allowed rapid conversion of these scaffolds to novel, polycyclic dihydroquinazolin-2-ones, 2-imino-1,3-benzothiazinanes, dihydroisoquinolin-3-ones and bridged tetrahydroquinolines. PMID:25267860

  4. Thermal design and development of actively cooled brushes for compact homopolar generators

    NASA Astrophysics Data System (ADS)

    Makel, D. B.

    1986-11-01

    The thermal and hydraulic design of actively cooled current transfer brushes for compact homopolar generators (HPG) is described. The development of high-energy-density HPG's at the Center for Electromechanics at the University of Texas at Austin requires brushgear capable of handling high current densities and large thermal loads. Platelet technology has been applied in the design of actively cooled brushes with coolant injection directly from the brush contact face into the brush-rotor interface. Coolant channels a few thousandths of an inch in diameter have been designed in brushes constructed of photoetched platelets of copper (0.020 to 0.005 in. thick) and then bonded to form the cooled brush. Platelet construction of brushes also permits the incorporation of internal instrumentation for temperature measurement. The brushes have been designed to provide data on the effects of rotor speed, current level, coolant flow rate, and coolant injection pattern.

  5. Compound control method of neutral point voltage of three level NPC inverter based on the vector method of virtual space and hysteresis control

    NASA Astrophysics Data System (ADS)

    Yang, Fengping; Xiao, Fangfei

    2017-03-01

    Current control methods include hardware control and software control corresponding to the inherent unbalance problem of neutral point voltage in three level NPC inverter. The hardware control is rarely used due to its high cost. In this paper, a new compound control method has been presented based on the vector method of virtual space and traditional hysteresis control of neutral point voltage, which can make up the shortcoming of the virtual control without the feedback control system of neutral point voltage and the blind area of hysteresis control and control the deviation and wave of neutral point voltage. The accuracy of this method has been demonstrated by simulation.

  6. Using Electronic Neutron Generators in Active Interrogation to Detect Shielded Fissionable Material

    SciTech Connect

    D. L. Chichester; E. H. Seabury

    2008-10-01

    Experiments have been performed at Idaho National Laboratory to study methodology and instrumentation for performing neutron active interrogation die-away analyses for the purpose of detecting shielded fissionable material. Here we report initial work using a portable DT electronic neutron generator with a He-3 fast neutron detector to detect shielded fissionable material including >2 kg quantities of enriched uranium and plutonium. Measurements have been taken of bare material as well as of material hidden within a large plywood cube. Results from this work have demonstrated the efficacy of the die-away neutron measurement technique for quickly detecting the presence of special nuclear material hidden within plywood shields by analyzing the time dependent neutron signals in-between neutron generator pulses. Using a DT electronic neutron generator operating at 300 Hz with a yield of approximately 0.36 x 10**8 neutrons per second, 2.2 kg of enriched uranium hidden within a 0.60 m x 0.60 m x 0.70 m volume of plywood was positively detected with a measurement signal 2-sigma above the passive background within 1 second. Similarly, for a 500 second measurement period a lower detection limit of approaching the gram level could be expected with the same simple set-up.

  7. Putative role of border cells in generating spontaneous morphological activity within Kölliker's organ.

    PubMed

    Dayaratne, M W Nishani; Vlajkovic, Srdjan M; Lipski, Janusz; Thorne, Peter R

    2015-12-01

    Kölliker's organ is a transient epithelial structure, comprising a major part of the organ of Corti during pre-hearing stages of development. The auditory system is spontaneously active during development, which serves to retain and refine neural connections. Kölliker's organ is considered a key candidate for generating such spontaneous activity, most likely through purinergic (P2 receptor) signalling and inner hair cell (IHC) activation. Associated with the spontaneous neural activity, ATP released locally by epithelial cells induces rhythmic morphological changes within Kölliker's organ, the purpose of which is not understood. These changes are accompanied by a shift in cellular refractive index, allowing optical detection of this activity in real-time. Using this principle, we investigated the origin of spontaneous morphological activity within Kölliker's organ. Apical turns of Wistar rat cochleae (P9-11) were dissected, and the purinergic involvement was studied following acute tissue exposure to a P2 receptor agonist (ATPγS) and antagonist (suramin). ATPγS induced a sustained darkening throughout Kölliker's organ, reversed by suramin. This effect was most pronounced in the region closest to the inner hair cells, which also displayed the highest frequency of intrinsic morphological events. Additionally, suramin alone induced swelling of this region, suggesting a tight regulation of cell volume by ATP-mediated mechanisms. Histological analysis of cochlear tissues demonstrates the most profound volume changes in the border cell region immediately adjacent to the IHCs. Together, these results underline the role of purinergic signalling in initiating morphological events within Kölliker's organ, and suggest a key involvement of border cells surrounding IHCs in regulating this spontaneous activity.

  8. A muscarinic cholinergic mechanism underlies activation of the central pattern generator for locust flight.

    PubMed

    Buhl, Edgar; Schildberger, Klaus; Stevenson, Paul A

    2008-07-01

    A central question in behavioural control is how central pattern generators (CPGs) for locomotion are activated. This paper disputes the key role generally accredited to octopamine in activating the CPG for insect flight. In deafferented locusts, fictive flight was initiated by bath application of the muscarinic agonist pilocarpine, the acetylcholine analogue carbachol, and the acetylcholinesterase blocker eserine, but not by nicotine. Furthermore, in addition to octopamine, various other amines including dopamine, tyramine and histamine all induced fictive flight, but not serotonin or the amine-precursor amino acid tyrosine. However, flight initiation was not reversibly blocked by aminergic antagonists, and was still readily elicited by both natural stimulation (wind) and pilocarpine in reserpinized, amine-depleted locusts. By contrast, the muscarinic antagonists atropine and scopolamine reversibly blocked flight initiated by wind, cholinergic agonists, octopamine, and by selective stimulation of a flight-initiating interneurone (TCG). The short delay from TCG stimulation to flight onset suggests that TCG acts directly on the flight CPG, and accordingly that TCG, or its follower cell within the flight generating circuit, is cholinergic. We conclude that acetylcholine acting via muscarinic receptors is the key neurotransmitter in the mechanism underlying the natural activation of the locust flight CPG. Amines are not essential for this, but must be considered as potential neuromodulators for facilitating flight release and tuning the motor pattern. We speculate that muscarinic activation coupled to aminergic facilitation may be a general feature of behavioural control in insects for ensuring conditional recruitment of individual motor programs in accordance with momentary adaptive requirements.

  9. Diosgenin inhibits superoxide generation in FMLP-activated mouse neutrophils via multiple pathways.

    PubMed

    Lin, Y; Jia, R; Liu, Y; Gao, Y; Zeng, X; Kou, J; Yu, B

    2014-12-01

    Diosgenin possesses anti-inflammatory and anticancer properties. Activated neutrophils produce high concentrations of the superoxide anion which is involved in the pathophysiology of inflammation-related diseases and cancer. In the present study, the inhibitory effect and possible mechanisms of diosgenin on superoxide generation were investigated in mouse bone marrow neutrophils. Diosgenin potently and concentration-dependently inhibited the extracellular and intracellular superoxide anion generation in Formyl-Met-Leu-Phe (FMLP)- activated neutrophils, with IC50 values of 0.50 ± 0.08 μM and 0.66 ± 0.13 μM, respectively. Such inhibition was not mediated by scavenging the superoxide anion or by a cytotoxic effect. Diosgenin inhibited the phosphorylation of p47phox and membrane translocation of p47phox and p67phox, and thus blocking the assembly of nicotinamide adenine dinucleotide phosphate oxidase. Moreover, cellular cyclic adenosine monophosphate (cAMP) levels and protein kinase A (PKA) expression were also effectively increased by diosgenin. It attenuated FMLP-induced increase of phosphorylation of cytosolic phospholipase A (cPLA2), p21-activated kinase (PAK), Akt, p38 mitogen-activated protein kinase (p38MAPK), extracellular signal-regulated kinase (ERK1/2), and c-Jun N-terminal kinase (JNK). Our data indicate that diosgenin exhibits inhibitory effects on superoxide anion production through the blockade of cAMP, PKA, cPLA2, PAK, Akt and MAPKs signaling pathways. The results may explain the clinical implications of diosgenin in the treatment of inflammation-related disorders.

  10. Ground-based acoustic parametric generator impact on the atmosphere and ionosphere in an active experiment

    NASA Astrophysics Data System (ADS)

    Rapoport, Yuriy G.; Cheremnykh, Oleg K.; Koshovy, Volodymyr V.; Melnik, Mykola O.; Ivantyshyn, Oleh L.; Nogach, Roman T.; Selivanov, Yuriy A.; Grimalsky, Vladimir V.; Mezentsev, Valentyn P.; Karataeva, Larysa M.; Ivchenko, Vasyl. M.; Milinevsky, Gennadi P.; Fedun, Viktor N.; Tkachenko, Eugen N.

    2017-01-01

    We develop theoretical basics of active experiments with two beams of acoustic waves, radiated by a ground-based sound generator. These beams are transformed into atmospheric acoustic gravity waves (AGWs), which have parameters that enable them to penetrate to the altitudes of the ionospheric E and F regions where they influence the electron concentration of the ionosphere. Acoustic waves are generated by the ground-based parametric sound generator (PSG) at the two close frequencies. The main idea of the experiment is to design the output parameters of the PSG to build a cascade scheme of nonlinear wave frequency downshift transformations to provide the necessary conditions for their vertical propagation and to enable penetration to ionospheric altitudes. The PSG generates sound waves (SWs) with frequencies f1 = 600 and f2 = 625 Hz and large amplitudes (100-420 m s-1). Each of these waves is modulated with the frequency of 0.016 Hz. The novelty of the proposed analytical-numerical model is due to simultaneous accounting for nonlinearity, diffraction, losses, and dispersion and inclusion of the two-stage transformation (1) of the initial acoustic waves to the acoustic wave with the difference frequency Δf = f2 - f1 in the altitude ranges 0-0.1 km, in the strongly nonlinear regime, and (2) of the acoustic wave with the difference frequency to atmospheric acoustic gravity waves with the modulational frequency in the altitude ranges 0.1-20 km, which then reach the altitudes of the ionospheric E and F regions, in a practically linear regime. AGWs, nonlinearly transformed from the sound waves, launched by the two-frequency ground-based sound generator can increase the transparency of the ionosphere for the electromagnetic waves in HF (MHz) and VLF (kHz) ranges. The developed theoretical model can be used for interpreting an active experiment that includes the PSG impact on the atmosphere-ionosphere system, measurements of electromagnetic and acoustic fields, study of

  11. Human cervical spinal cord circuitry activated by tonic input can generate rhythmic arm movements.

    PubMed

    Solopova, I A; Selionov, V A; Zhvansky, D S; Gurfinkel, V S; Ivanenko, Y

    2016-02-01

    The coordination between arms and legs during human locomotion shares many features with that in quadrupeds, yet there is limited evidence for the central pattern generator for the upper limbs in humans. Here we investigated whether different types of tonic stimulation, previously used for eliciting stepping-like leg movements, may evoke nonvoluntary rhythmic arm movements. Twenty healthy subjects participated in this study. The subject was lying on the side, the trunk was fixed, and all four limbs were suspended in a gravity neutral position, allowing unrestricted low-friction limb movements in the horizontal plane. The results showed that peripheral sensory stimulation (continuous muscle vibration) and central tonic activation (postcontraction state of neuronal networks following a long-lasting isometric voluntary effort, Kohnstamm phenomenon) could evoke nonvoluntary rhythmic arm movements in most subjects. In ∼40% of subjects, tonic stimulation elicited nonvoluntary rhythmic arm movements together with rhythmic movements of suspended legs. The fact that not all participants exhibited nonvoluntary limb oscillations may reflect interindividual differences in responsiveness of spinal pattern generation circuitry to its activation. The occurrence and the characteristics of induced movements highlight the rhythmogenesis capacity of cervical neuronal circuitries, complementing the growing body of work on the quadrupedal nature of human gait.

  12. Thrombin-activated human platelets acutely generate oxidized docosahexaenoic-acid-containing phospholipids via 12-lipoxygenase.

    PubMed

    Morgan, Lloyd T; Thomas, Christopher P; Kühn, Hartmut; O'Donnell, Valerie B

    2010-10-01

    Arachidonate-containing oxidized phospholipids are acutely generated by 12-LOX (12-lipoxygenase) in agonist-activated platelets. In the present study, formation of structurally related lipids by oxidation of DHA (docosahexaenoic acid)-containing phospholipids is demonstrated using lipidomic approaches. Precursor scanning reverse-phase LC (liquid chromatography)-MS/MS (tandem MS) identified a new family of lipids that comprise phospholipid-esterified HDOHE (hydroxydocosahexaenoic acid). Two diacyl and two plasmalogen PEs (phosphatidylethanolamines) containing predominantly the 14-HDOHE positional isomer (18:0p/14-HDOHE-PE, 18:0a/14-HDOHE-PE, 16:0a/14-HDOHE-PE and 16:0p/14-HDOHE-PE) were structurally characterized using MS/MS and by comparison with biogenic standards. An involvement of 12-LOX was indicated as purified recombinant human 12-LOX also generated the 14-HDOHE isomer from DHA. Pharmacological studies using inhibitors and recombinant platelet 12-LOX indicate that they form via esterification of newly formed non-esterified HDOHE. HDOHE-PEs formed at significant rates (2-4 ng/4×10(7) cells) within 2-180 min of thrombin stimulation, and their formation was blocked by calcium chelation. In summary, a new family of oxidized phospholipid was identified in thrombin-activated human platelets.

  13. Procyclic Trypanosoma brucei do not use Krebs cycle activity for energy generation.

    PubMed

    van Weelden, Susanne W H; Fast, Beate; Vogt, Achim; van der Meer, Pieter; Saas, Joachim; van Hellemond, Jaap J; Tielens, Aloysius G M; Boshart, Michael

    2003-04-11

    The importance of a functional Krebs cycle for energy generation in the procyclic stage of Trypanosoma brucei was investigated under physiological conditions during logarithmic phase growth of a pleomorphic parasite strain. Wild type procyclic cells and mutants with targeted deletion of the gene coding for aconitase were derived by synchronous in vitro differentiation from wild type and mutant (Delta aco::NEO/Delta aco::HYG) bloodstream stage parasites, respectively, where aconitase is not expressed and is dispensable. No differences in intracellular levels of glycolytic and Krebs cycle intermediates were found in procyclic wild type and mutant cells, except for citrate that accumulated up to 90-fold in the mutants, confirming the absence of aconitase activity. Surprisingly, deletion of aconitase did not change differentiation nor the growth rate or the intracellular ATP/ADP ratio in those cells. Metabolic studies using radioactively labeled substrates and NMR analysis demonstrated that glucose and proline were not degraded via the Krebs cycle to CO(2). Instead, glucose was degraded to acetate, succinate, and alanine, whereas proline was degraded to succinate. Importantly, there was absolutely no difference in the metabolic products released by wild type and aconitase knockout parasites, and both were for survival strictly dependent on respiration via the mitochondrial electron transport chain. Hence, although the Krebs cycle enzymes are present, procyclic T. brucei do not use Krebs cycle activity for energy generation, but the mitochondrial respiratory chain is essential for survival and growth. We therefore propose a revised model of the energy metabolism of procyclic T. brucei.

  14. Activity-Dependent p25 Generation Regulates Synaptic Plasticity and Aβ-Induced Cognitive Impairment

    PubMed Central

    Seo, Jinsoo; Giusti-Rodríguez, Paola; Zhou, Ying; Rudenko, Andrii; Cho, Sukhee; Ota, Kristie T.; Park, Christine; Patzke, Holger; Madabhushi, Ram; Pan, Ling; Mungenast, Alison E.; Guan, Ji-Song; Delalle, Ivana; Tsai, Li-Huei

    2015-01-01

    SUMMARY Cyclin-dependent kinase 5 regulates numerous neuronal functions with its activator, p35. Under neurotoxic conditions, p35 undergoes proteolytic cleavage to liberate p25, which has been implicated in various neurodegenerative diseases. Here, we show that p25 is generated following neuronal activity under physiological conditions in a GluN2B- and CaMKIIα-dependent manner. Moreover, we developed a knockin mouse model in which endogenous p35 is replaced with a calpain-resistant mutant p35 (Δp35KI) to prevent p25 generation. The Δp35KI mice exhibit impaired long-term depression and defective memory extinction, likely mediated through persistent GluA1 phosphorylation at Ser845. Finally, crossing the Δp35KI mice with the 5XFAD mouse model of Alzheimer's disease (AD) resulted in an amelioration of β-amyloid (Aβ)-induced synaptic depression and cognitive impairment. Together, these results reveal a physiological role of p25 production in synaptic plasticity and memory and provide new insights into the function of p25 in Aβ-associated neurotoxicity and AD-like pathology. PMID:24725413

  15. A NOVEL MITHRAMYCIN ANALOGUE WITH HIGH ANTITUMOR ACTIVITY AND LESS TOXICITY GENERATED BY COMBINATORIAL BIOSYNTHESIS

    PubMed Central

    Núñez, Luz E.; Nybo, Stephen E.; González-Sabín, Javier; Pérez, María; Menéndez, Nuria; Braña, Alfredo F.; He, Min; Morís, Francisco; Salas, José A.; Rohr, Jürgen; Méndez, Carmen

    2012-01-01

    Mithramycin is an antitumor compound produced by Streptomyces argillaceus that has been used for the treatment of several types of tumors and hypercalcaemia processes. However, its use in humans has been limited because its side effects. Using combinatorial biosynthesis approaches, we have generated seven new mithramycin derivatives, which either differ from the parental compound in the sugar profile or in both the sugar profile and the 3-side chain. From these studies three novel derivatives were identified, demycarosyl-3D-β-d-digitoxosyl-mithramycin SK, demycarosyl-mithramycin SDK and demycarosyl-3D-β-d-digitoxosyl-mithramycin SDK, which show high antitumor activity. The first one, which combines two structural features previously found to improve pharmacological behavior, was generated following two different strategies, and it showed less toxicity than mithramycin. Preliminary in vivo evaluation of its antitumor activity through hollow fiber assays, and in subcutaneous colon and melanoma cancers xenografts models, suggests that demycarosyl-3D-β-d-digitoxosyl-mithramycin SK could be a promising antitumor agent, worthy of further investigation. PMID:22578073

  16. Acid sphingomyelinase is activated in sickle cell erythrocytes and contributes to inflammatory microparticle generation in SCD.

    PubMed

    Awojoodu, Anthony O; Keegan, Philip M; Lane, Alicia R; Zhang, Yuying; Lynch, Kevin R; Platt, Manu O; Botchwey, Edward A

    2014-09-18

    Sphingolipids are a class of lipids containing a backbone of sphingoid bases that can be produced de novo through the reaction of palmitate and serine and further metabolized through the activity of various enzymes to produce intermediates with diverse roles in cellular processes and signal transduction. One of these intermediates, sphingosine 1-phosphate (S1P), is stored at high concentrations (1 μM) in red blood cells (RBCs) and directs a wide array of cellular processes mediated by 5 known G-protein coupled receptors (S1P1-S1P5). In this study, we show that RBC membrane alterations in sickle cell disease enhance the activation acid sphingomyelinase by 13%, resulting in increased production and storage of sphingosine (2.6-fold) and S1P (3.5-fold). We also show that acid sphingomyelinase enhances RBC-derived microparticle (MP) generation. These MPs are internalized by myeloid cells and promote proinflammatory cytokine secretion and endothelial cell adhesion, suggesting that potential crosstalk between circulating inflammatory cells and MPs may contribute to the inflammation-rooted pathogenesis of the disease. Treatment with amitriptyline reduces MP generation in vitro and in vivo and might be used to mitigate inflammatory processes in sickle cell disease.

  17. Increased AICD generation does not result in increased nuclear translocation or activation of target gene transcription

    SciTech Connect

    Waldron, Elaine; Isbert, Simone; Kern, Andreas; Jaeger, Sebastian; Martin, Anne M.; Hebert, Sebastien S.; Behl, Christian; Weggen, Sascha; De Strooper, Bart; Pietrzik, Claus U.

    2008-08-01

    A sequence of amyloid precursor protein (APP) cleavages culminates in the sequential release of the APP intracellular domain (AICD) and the amyloid {beta} peptide (A{beta}) and/or p3 fragment. One of the environmental factors favouring the accumulation of AICD appears to be a rise in intracellular pH. Here we further identified the metabolism and subcellular localization of artificially expressed constructs under such conditions. We also co-examined the mechanistic lead up to the AICD accumulation and explored possible significances for its increased expression. We found that most of the AICD generated under pH neutralized conditions is likely cleaved from C83. While the AICD surplus was unable to further activate transcription of a luciferase reporter via a Gal4-DNA-binding domain, it failed entirely via the endogenous promoter regions of proposed target genes, APP and KAI1. The lack of a specific transactivation potential was also demonstrated by the unchanged levels of target gene mRNA. However, rather than translocating to the nucleus, the AICD surplus remains membrane tethered or free in the cytosol where it interacts with Fe65. Therefore we provide strong evidence that an increase in AICD generation does not directly promote gene activation of previously proposed target 0011gen.

  18. Loss of contractile activity of endothelin-1 induced by electrical field stimulation-generated free radicals.

    PubMed

    Yasuda, N; Kasuya, Y; Yamada, G; Hama, H; Masaki, T; Goto, K

    1994-09-01

    1. Electrical field stimulation (EFS; 10 V, 10 Hz, 2 ms) of porcine coronary artery strips precontracted with 10 nM endothelin-1 (ET-1) for 5 min caused a biphasic response, consisting of a slight contraction during EFS and a marked and irreversible relaxation just after EFS. This irreversible relaxation after EFS has never been investigated. In the present study, we have investigated the mechanism of the relaxation after EFS. 2. The EFS-induced response was not affected by the presence or absence of endothelium and was insensitive to 10 microM tetrodotoxin (TTX). 3. In the presence of free radical scavengers (40 u ml-1 superoxide dismutase (SOD), 1200 u ml-1 catalase or 80 mM D-mannitol), the relaxation after EFS was significantly inhibited. Moreover, relaxation after EFS was not observed in porcine coronary artery strips precontracted with 20 mM KCl. 4. In a cascade experiment, EFS of Krebs-Ringer solution containing 10 nM ET-1 induced marked suppression of the contractile activity of ET-1 in porcine coronary artery strips, which was in accord with the observed decrease in release of immunoreactive ET-1 (ir-ET-1). This effect of EFS was significantly inhibited by each of the free radical scavengers, 3 mM vitamin C, 40 u ml-1 SOD, 1200 u ml-1 catalase and 80 mM D-mannitol. 5. The exchange of 95% O2/5% CO2 gas for 95% N2/5% CO2 gas significantly inhibited the EFS-induced decrease in release of ir-ET-1. 6. Neither superoxide anions generated by xanthine (10 JM) plus xanthine oxidase (0.1 micro ml-1) nor hydrogen peroxide (10 microM) exogenously added to Krebs-Ringer solution containing 10 nM ET-1 affected the level of ir-ET-1.7. Generation of hydroxyl radicals was detected in the EFS-applied Krebs-Ringer solution. The EFS-induced generation of hydroxyl radicals was dependent on the period of stimulation and 02-bubbling, and significant generation of hydroxyl radicals was detectable with stimulation of over 5 min.Moreover, hydroxyl radicals generated in 50 mM Na

  19. Hydrogenase activity and proton-motive force generation by Escherichia coli during glycerol fermentation.

    PubMed

    Trchounian, Karen; Blbulyan, Syuzanna; Trchounian, Armen

    2013-06-01

    Proton motive force (Δp) generation by Escherichia coli wild type cells during glycerol fermentation was first studied. Its two components, electrical-the membrane potential (∆φ) and chemical-the pH transmembrane gradient (ΔpH), were established and the effects of external pH (pHex) were determined. Intracellular pH was 7.0 and 6.0 and lower than pHex at pH 7.5 and 6.5, respectively; and it was higher than pHex at pH 5.5. At high pHex, the increase of ∆φ (-130 mV) was only partially compensated by a reversed ΔpH, resulting in a low Δp. At low pHex ∆φ and consequently Δp were decreased. The generation of Δp during glycerol fermentation was compared with glucose fermentation, and the difference in Δp might be due to distinguished mechanisms for H(+) transport through the membrane, especially to hydrogenase (Hyd) enzymes besides the F0F1-ATPase. H(+) efflux was determined to depend on pHex; overall and N,N'-dicyclohexylcarbodiimide (DCCD)-inhibitory H(+) efflux was maximal at pH 6.5. Moreover, ΔpH was changed at pH 6.5 and Δp was different at pH 6.5 and 5.5 with the hypF mutant lacking all Hyd enzymes. DCCD-inhibited ATPase activity of membrane vesicles was maximal at pH 7.5 and decreased with the hypF mutant. Thus, Δp generation by E. coli during glycerol fermentation is different than that during glucose fermentation. Δp is dependent on pHex, and a role of Hyd enzymes in its generation is suggested.

  20. Advanced Glycation End Products Activate a Chymase-Dependent Angiotensin II Generating Pathway in Diabetic Complications

    PubMed Central

    Koka, Vijay; Wang, Wansheng; Huang, Xiao Ru; Kim-Mitsuyama, Shokei; Truong, Luan D.; Lan, Hui Y

    2006-01-01

    Background: Angiotensin II is a key mediator of diabetes-related vascular disease. It is now recognized that in addition to angiotensin converting enzyme (ACE), chymase is an important alternative angiotensin II generating enzyme in hypertension and diabetes. However, the mechanism of induction of chymase in diabetes remains unknown. Methods and Results: Here we report that chymase is upregulated in coronary and renal arteries in patients with diabetes by immunohistochemistry. Upregulation of vascular chymase is associated with deposition of advanced glycation end products (AGEs), increase in expression of the receptor for AGEs (RAGE), and activation of ERK1/2 MAP kinase. In vitro, AGEs can induce chymase expression and chymase-dependent angiotensin II generation in human vascular smooth muscle cells via the RAGE-ERK1/2 MAP kinase-dependent mechanism. This is confirmed by blockade of AGE-induced vascular chymase expression with a neutralizing RAGE antibody and an inhibitor to ERK1/2, and by overexpression of the dominant negative-ERK1/2. Compared to ACE, chymase contributes to the majority of angiotensin II production (more than 70%, p<0.01) in response to AGEs. Further more, AGE-induced Angiotensin II production is blocked by the anti-RAGE antibody and by inhibition of ERK1/2 MAP kinase activities. Conclusions: Advanced glycation end products, a hallmark of diabetes, induce chymase via the RAGE-ERK1/2 MAP kinase pathway. Chymase initiates an important alternative angiotensin II generating pathway in diabetes and may play a critical role in diabetic vascular disease. PMID:16520412

  1. A new generation active arrays for optical flexibility in astronomical instrumentation

    NASA Astrophysics Data System (ADS)

    Kroes, G.; Jaskó, A.; Pragt, J. H.; Venema, L.; De Haan, M.

    2012-09-01

    Throughout the history of telescopes and astronomical instrumentation, new ways were found to open up unexplored possibilities in fundamental astronomical research by increasing the telescope size and instrumentation complexity. The ever demanding requirements on instrument performance pushes instrument complexity to the edge. In order to take the next leap forward in instrument development the optical design freedom needs to be increased drastically. The use of more complex and more accurate optics allows for shorter optical trains with smaller sizes, smaller number of components and reduced fabrication and alignment verification time and costs. Current optics fabrication is limited in surface form complexity and/or accuracy. Traditional active and adaptive optics lack the needed intrinsic long term stability and simplicity in design, manufacturing, verification and control. This paper explains how and why active arrays literally provide a flexible but stable basis for the next generation optical instruments. Combing active arrays with optically high quality face sheets more complex and accurate optical surface forms can be provided including extreme a-spherical (freeform) surfaces and thus allow for optical train optimization and even instrument reconfiguration. A zero based design strategy is adopted for the development of the active arrays addressing fundamental issues in opto-mechanical engineering. The various choices are investigated by prototypes and Finite Element Analysis. Finally an engineering concept will be presented following a highly stable adjustment strategy allowing simple verification and control. The Optimization metrology is described in an additional paper for this conference by T. Agócs et al.

  2. Generation of reproducible turbulent inflows for wind tunnel applications using active grids

    NASA Astrophysics Data System (ADS)

    Kroeger, Lars; Guelker, Gerd; Peinke, Joachim

    2016-11-01

    Turbulent flows are omnipresent in nature. In the case of wind energy applications, reproducible measurements in situ are quite difficult, therefore research in turbulence demands for experimental setups with reproducible turbulent flow fields. To simulate the situation from the outside in a wind tunnel an active grid can be used. It consists of horizontal and vertical rotating axes with attached square flaps which could be moved individually. This dynamically driven setup and the possibility to repeat the motions of the active grid axes permits to generate reproducible, statistically well defined turbulence with a wide range of statistical behavior. The objective of this work is to create turbulence with two active grids of different dimensions, to establish comparable setups in our available wind tunnel facilities. In this study the wake of the active grids was investigated by high speed PIV and hotwire measurements. To determine the similarities and limitations between the setups of different dimensions the hotwire data is compared using higher order statistics, increment analysis and the power spectra. The PIV data is used to observe spatial correlations and the prevailing length scales in the turbulent wakes. First results regarding this comparison are shown.

  3. Generation of structurally novel short carotenoids and study of their biological activity

    PubMed Central

    Kim, Se H.; Kim, Moon S.; Lee, Bun Y.; Lee, Pyung C.

    2016-01-01

    Recent research interest in phytochemicals has consistently driven the efforts in the metabolic engineering field toward microbial production of various carotenoids. In spite of systematic studies, the possibility of using C30 carotenoids as biologically functional compounds has not been explored thus far. Here, we generated 13 novel structures of C30 carotenoids and one C35 carotenoid, including acyclic, monocyclic, and bicyclic structures, through directed evolution and combinatorial biosynthesis, in Escherichia coli. Measurement of radical scavenging activity of various C30 carotenoid structures revealed that acyclic C30 carotenoids showed higher radical scavenging activity than did DL-α-tocopherol. We could assume high potential biological activity of the novel structures of C30 carotenoids as well, based on the neuronal differentiation activity observed for the monocyclic C30 carotenoid 4,4′-diapotorulene on rat bone marrow mesenchymal stem cells. Our results demonstrate that a series of structurally novel carotenoids possessing biologically beneficial properties can be synthesized in E. coli. PMID:26902326

  4. We are what we do: Examining learner-generated content in the anatomy laboratory through the lens of activity theory.

    PubMed

    Doubleday, Alison F; Wille, Sarah J

    2014-01-01

    Video and photography are often used for delivering content within the anatomical sciences. However, instructors typically produce these resources to provide instructional or procedural information. Although the benefits of learner-generated content have been explored within educational research, virtually no studies have investigated the use of learner-generated video and photograph content within anatomy dissection laboratories. This study outlines an activity involving learner-generated video diaries and learner-generated photograph assignments produced during anatomy laboratory sessions. The learner-generated photographs and videos provided instructors with a means of formative assessment and allowed instructors to identify evidence of collaborative behavior in the laboratory. Student questionnaires (n = 21) and interviews (n = 5), as well as in-class observations, were conducted to examine student perspectives on the laboratory activities. The quantitative and qualitative data were examined using the framework of activity theory to identify contradictions between student expectations of, and engagement with, the activity and the actual experiences of the students. Results indicate that learner-generated photograph and video content can act as a rich source of data on student learning processes and can be used for formative assessment, for observing collaborative behavior, and as a starting point for class discussions. This study stresses the idea that technology choice for activities must align with instructional goals. This research also highlights the utility of activity theory as a framework for assessing classroom and laboratory activities, demonstrating that this approach can guide the development of laboratory activities.

  5. ROS-generating/ARE-activating capacity of metals in roadway particulate matter deposited in urban environment.

    PubMed

    Shuster-Meiseles, Timor; Shafer, Martin M; Heo, Jongbae; Pardo, Michal; Antkiewicz, Dagmara S; Schauer, James J; Rudich, Assaf; Rudich, Yinon

    2016-04-01

    In this study we investigated the possible causal role for soluble metal species extracted from roadway traffic emissions in promoting particulate matter (PM)-induced reactive oxygen species (ROS) production and antioxidant response element (ARE) promoter activation. To this end, these responses have been evaluated in alveolar macrophage and epithelial lung cells that have been exposed to 'Unfiltered', 'Filtered' and 'Filtered+Chelexed' water extracts of PM samples collected from the roadway urban environments of Thessaloniki, Milan and London. Except for Thessaloniki, our results demonstrate that filtration resulted in a minor decrease in ROS activity of the fine PM fraction, suggesting that ROS activity is attributed mainly to water-soluble PM species. In contrast to ROS, ARE activity was mediated predominantly by the water-soluble component of PM present in both the fine and coarse extracts. Further removal of metals by Chelex treatment from filtered water extracts showed that soluble metal species are the major factors mediating ROS and ARE activities of the soluble fraction, especially in the London PM extracts. Finally, utilizing step-wise multiple-regression analysis, we show that 87% and 78% of the total variance observed in ROS and ARE assays, respectively, is accounted for by changes in soluble metal concentration. Using a statistical analysis we find that As, Zn and Fe best predict the ROS-generating/ARE-activating capacity of the near roadway particulate matter in the pulmonary cells studied. Collectively, our findings imply that soluble metals present in roadside PM are potential drivers of both pro- and anti-oxidative effects of PM in respiratory tract.

  6. Anesthetic Isoflurane Increases Phosphorylated Tau Levels Mediated by Caspase Activation and Aβ Generation

    PubMed Central

    Dong, Yuanlin; Wu, Xu; Xu, Zhipeng; Zhang, Yiying; Xie, Zhongcong

    2012-01-01

    Anesthetic isoflurane has been shown to promote Alzheimer’s disease (AD) neuropathogenesis by inducing caspase activation and accumulation of β-amyloid (Aβ). Phosphorylation of tau protein is another important feature of AD neuropathogenesis. However, the effects of isoflurane on phosphorylated tau levels remain largely to be determined. We therefore set out to determine whether isoflurane can increase phosphorylated tau levels. 5 to 8 month-old wild-type and AD transgenic mice [B6.Cg-Tg (APPswe, PSEN1dE9)85Dbo/J] were treated with 1.4% isoflurane for two hours. The mice brain tissues were harvested at six, 12 and 24 hours after the anesthesia. For the in vitro studies, primary neurons from wild-type and the AD transgenic mice were exposed to 2% isoflurane for six hours, and were harvested at the end of anesthesia. The harvested brain tissues and neurons were subjected to Western blot analysis by which the levels of phosphorylated tau protein at Serine 262 (Tau-PS262) were determined. Here we show that the isoflurane anesthesia increased Tau-PS262 levels in brain tissues and primary neurons from the wild-type and AD transgenic mice. Moreover, the isoflurane anesthesia may induce a greater increase in Tau-PS262 levels in primary neurons and brain tissues from the AD transgenic mice. Finally, caspase activation inhibitor Z-VAD and Aβ generation inhibitor L-685,458 attenuated the isoflurane-induced increases in Tau-PS262 levels. In conclusion, clinically relevant isoflurane anesthesia increases phosphorylated tau levels, which may result from the isoflurane-induced caspase activation and Aβ generation. These findings will promote more studies to determine the effects of anesthetics on tau phosphorylation. PMID:22745746

  7. Efficient generation of gene-modified pigs via injection of zygote with Cas9/sgRNA.

    PubMed

    Wang, Yong; Du, Yinan; Shen, Bin; Zhou, Xiaoyang; Li, Jian; Liu, Yu; Wang, Jianying; Zhou, Jiankui; Hu, Bian; Kang, Nannan; Gao, Jimin; Yu, Liqing; Huang, Xingxu; Wei, Hong

    2015-02-05

    Co-injection of zygotes with Cas9 mRNA and sgRNA has been proven to be an efficient gene-editing strategy for genome modification of different species. Genetic engineering in pigs holds a great promise in biomedical research. By co-injection of one-cell stage embryos with Cas9 mRNA and Npc1l1 sgRNA, we achieved precise Npc1l1 targeting in Chinese Bama miniature pigs at the efficiency as high as 100%. Meanwhile, we carefully analyzed the Npc1l1 sgRNA:Cas9-mediated on- and off-target mutations in various somatic tissues and ovaries, and demonstrated that injection of zygotes with Cas9 mRNA and sgRNA is an efficient and reliable approach for generation of gene-modified pigs.

  8. 40 CFR 761.64 - Disposal of wastes generated as a result of research and development activities authorized under...

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 40 Protection of Environment 32 2012-07-01 2012-07-01 false Disposal of wastes generated as a result of research and development activities authorized under § 761.30(j) and chemical analysis of PCBs..., AND USE PROHIBITIONS Storage and Disposal § 761.64 Disposal of wastes generated as a result...

  9. 40 CFR 761.64 - Disposal of wastes generated as a result of research and development activities authorized under...

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 40 Protection of Environment 32 2013-07-01 2013-07-01 false Disposal of wastes generated as a result of research and development activities authorized under § 761.30(j) and chemical analysis of PCBs..., AND USE PROHIBITIONS Storage and Disposal § 761.64 Disposal of wastes generated as a result...

  10. The Resveratrol Trimer Miyabenol C Inhibits β-Secretase Activity and β-Amyloid Generation

    PubMed Central

    Hu, Jin; Lin, Ting; Gao, Yuehong; Xu, Junyue; Jiang, Chao; Wang, Guanghui; Bu, Guojun; Xu, Huaxi; Chen, Haifeng; Zhang, Yun-wu

    2015-01-01

    Accumulation and deposition of amyloid-β peptide (Aβ) in the brain is a primary cause of the pathogenesis of Alzheimer’s disease (AD). Aβ is generated from amyloid-β precursor protein (APP) through sequential cleavages first by β-secretase and then by γ-secretase. Inhibiting β-secretase activity is believed to be one of the most promising strategies for AD treatment. In the present study, we found that a resveratrol trimer, miyabenol C, isolated from stems and leaves of the small-leaf grape (Vitisthunbergii var. taiwaniana), can markedly reduce Aβ and sAPPβ levels in both cell cultures and the brain of AD model mice. Mechanistic studies revealed that miyabenol C affects neither protein levels of APP, the two major α-secretases ADAM10 and TACE, and the γ-secretase component Presenilin 1, nor γ-secretase-mediated Notch processing and TACE activity. In contrast, although miyabenol C has no effect on altering protein levels of the β-secretase BACE1, it can inhibit both in vitro and in vivo β-secretase activity. Together, our results indicate that miyabenol C is a prominent β-secretase inhibitor and lead compound for AD drug development. PMID:25629409

  11. Gene Replacement for the Generation of Designed Novel Avermectin Derivatives with Enhanced Acaricidal and Nematicidal Activities

    PubMed Central

    Huang, Jun; Chen, An-Liang; Zhang, Hui; Yu, Zhen; Li, Mei-Hong; Li, Na; Lin, Jia-Tan; Bai, Hua

    2015-01-01

    Avermectin (AVM) and ivermectin (IVM) are potent pesticides and acaricides which have been widely used during the past 30 years. As insect resistance to AVM and IVM is greatly increasing, alternatives are urgently needed. Here, we report two novel AVM derivatives, tenvermectin A (TVM A) and TVM B, which are considered a potential new generation of agricultural and veterinary drugs. The molecules of the TVMs were designed based on structure and pharmacological property comparisons among AVM, IVM, and milbemycin (MBM). To produce TVMs, a genetically engineered strain, MHJ1011, was constructed from Streptomyces avermitilis G8-17, an AVM industrial strain. In MHJ1011, the native aveA1 gene was seamlessly replaced with milA1 from Streptomyces hygroscopicus. The total titer of the two TVMs produced by MHJ1011 reached 3,400 mg/liter. Insecticidal tests proved that TVM had enhanced activities against Tetranychus cinnabarinus and Bursaphelenchus xylophilus, as desired. This study provides a typical example of exploration for novel active compounds through a new method of polyketide synthase (PKS) reassembly for gene replacement. The results of the insecticidal tests may be of use in elucidating the structure-activity relationship of AVMs and MBMs. PMID:26025902

  12. Phenol degradation in heterogeneous system generating singlet oxygen employing light activated electropolymerized phenothiazines

    NASA Astrophysics Data System (ADS)

    Piwowar, Katarzyna; Blacha-Grzechnik, Agata; Bernas, Paulina; Zak, Jerzy

    2015-12-01

    Five selected amine-derivatives of phenothiazine were electropolymerized on an ITO/glass substrate and then used in the daylight-activated process to produce in situ singlet oxygen which degrades phenol in a solution. The phenothiazines were immobilized in a simple electrochemical procedure in an acidic solution which led to the formation of an ultrathin transparent polymeric film. All films obtained on the ITO substrate including azure A (AA), azure C (AC), methylene blue (MB), toluidine blue (TBO), and thionine (Th) had a comparable surface coverage at the level of picomoles/cm2. The activity of these materials was then compared and presented in terms of an efficiency of the phenol degradation process in an aqueous solution by photogenerated singlet oxygen. That efficiency was determined by the UV-vis spectroscopy employing a phenol/4-aminoantipyrine complex. All the phenothiazine ultrathin polymeric films were capable of generating the singlet oxygen in the aqueous solution under daylight activation, which was used in the consecutive process of phenol degradation. The highest efficiency at a level of 51.4% and 45.4% was found for the AC/ITO and MB/ITO layers, respectively.

  13. Simplified 2D Bidomain Model of Whole Heart Electrical Activity and ECG Generation

    NASA Astrophysics Data System (ADS)

    Sovilj, Siniša; Magjarević, Ratko; Abed, Amr Al; Lovell, Nigel H.; Dokos, Socrates

    2014-06-01

    The aim of this study was the development of a geometrically simple and highly computationally-efficient two dimensional (2D) biophysical model of whole heart electrical activity, incorporating spontaneous activation of the sinoatrial node (SAN), the specialized conduction system, and realistic surface ECG morphology computed on the torso. The FitzHugh-Nagumo (FHN) equations were incorporated into a bidomain finite element model of cardiac electrical activity, which was comprised of a simplified geometry of the whole heart with the blood cavities, the lungs and the torso as an extracellular volume conductor. To model the ECG, we placed four electrodes on the surface of the torso to simulate three Einthoven leads VI, VII and VIII from the standard 12-lead system. The 2D model was able to reconstruct ECG morphology on the torso from action potentials generated at various regions of the heart, including the sinoatrial node, atria, atrioventricular node, His bundle, bundle branches, Purkinje fibers, and ventricles. Our 2D cardiac model offers a good compromise between computational load and model complexity, and can be used as a first step towards three dimensional (3D) ECG models with more complex, precise and accurate geometry of anatomical structures, to investigate the effect of various cardiac electrophysiological parameters on ECG morphology.

  14. Gene Replacement for the Generation of Designed Novel Avermectin Derivatives with Enhanced Acaricidal and Nematicidal Activities.

    PubMed

    Huang, Jun; Chen, An-Liang; Zhang, Hui; Yu, Zhen; Li, Mei-Hong; Li, Na; Lin, Jia-Tan; Bai, Hua; Wang, Ji-Dong; Zheng, Yu-Guo

    2015-08-15

    Avermectin (AVM) and ivermectin (IVM) are potent pesticides and acaricides which have been widely used during the past 30 years. As insect resistance to AVM and IVM is greatly increasing, alternatives are urgently needed. Here, we report two novel AVM derivatives, tenvermectin A (TVM A) and TVM B, which are considered a potential new generation of agricultural and veterinary drugs. The molecules of the TVMs were designed based on structure and pharmacological property comparisons among AVM, IVM, and milbemycin (MBM). To produce TVMs, a genetically engineered strain, MHJ1011, was constructed from Streptomyces avermitilis G8-17, an AVM industrial strain. In MHJ1011, the native aveA1 gene was seamlessly replaced with milA1 from Streptomyces hygroscopicus. The total titer of the two TVMs produced by MHJ1011 reached 3,400 mg/liter. Insecticidal tests proved that TVM had enhanced activities against Tetranychus cinnabarinus and Bursaphelenchus xylophilus, as desired. This study provides a typical example of exploration for novel active compounds through a new method of polyketide synthase (PKS) reassembly for gene replacement. The results of the insecticidal tests may be of use in elucidating the structure-activity relationship of AVMs and MBMs.

  15. Active Mediation of Plasmon Enhanced Localized Exciton Generation, Carrier Diffusion and Enhanced Photon Emission.

    PubMed

    Haq, Sharmin; Addamane, Sadhvikas; Kafle, Bijesh; Huang, Danhong; Balakrishnan, Ganesh; Habteyes, Terefe G

    2017-04-13

    Understanding the enhancement of charge carrier generation and their diffusion is imperative for improving the efficiency of optoelectronic devices particularly infrared photodetectors that are less developed than their visible counterpart. Here, using gold nanorods as model plasmonic systems, InAs quantum dots (QDs) embedded in an InGaAs quantum well as an emitter, and GaAs as an active mediator of surface plasmons for enhancing carrier generation and photon emission, the distance dependence of energy transfer and carrier diffusion have been investigated both experimentally and theoretically. Analysis of the QD emission enhancement as a function of distance reveals a Förster radius of 3.85 ± 0.15 nm, a near-field decay length of 4.8 ± 0.1 nm and an effective carrier diffusion length of 64.0 ± 3.0 nm. Theoretical study of the temporal-evolution of the electron-hole occupation number of the excited states of the QDs indicates that the emission enhancement trend is determined by the carrier diffusion and capture rates.

  16. GABAergic synchronization in the limbic system and its role in the generation of epileptiform activity

    PubMed Central

    Avoli, Massimo; de Curtis, Marco

    2016-01-01

    GABA is the main inhibitory neurotransmitter in the adult forebrain, where it activates ionotropic type A and metabotropic type B receptors. Early studies have shown that GABAA receptor-mediated inhibition controls neuronal excitability and thus the occurrence of seizures. However, more complex, and at times unexpected, mechanisms of GABAergic signaling have been identified during epileptiform discharges over the last few years. Here, we will review experimental data that point at the paradoxical role played by GABAA receptor-mediated mechanisms in synchronizing neuronal networks, and in particular those of limbic structures such as the hippocampus, the entorhinal and perirhinal cortices, or the amygdala. After having summarized the fundamental characteristics of GABAA receptor-mediated mechanisms, we will analyze their role in the generation of network oscillations and their contribution to epileptiform synchronization. Whether and how GABAA receptors influence the interaction between limbic networks leading to ictogenesis will be also reviewed. Finally, we will consider the role of altered inhibition in the human epileptic brain along with the ability of GABAA receptor-mediated conductances to generate synchronous depolarizing events that may lead to ictogenesis in human epileptic disorders as well. PMID:21802488

  17. Acoustic manipulation of active spherical carriers: Generation of negative radiation force

    NASA Astrophysics Data System (ADS)

    Rajabi, Majid; Mojahed, Alireza

    2016-09-01

    This paper examines theoretically a novel mechanism of generating negative (pulling) radiation force for acoustic manipulation of spherical carriers equipped with piezoelectric actuators in its inner surface. In this mechanism, the spherical particle is handled by common plane progressive monochromatic acoustic waves instead of zero-/higher- order Bessel beams or standing waves field. The handling strategy is based on applying a spatially uniform harmonic electrical voltage at the piezoelectric actuator with the same frequency of handling acoustic waves, in order to change the radiation force effect from repulsive (away from source) to attractive (toward source). This study may be considered as a start point for development of contact-free precise handling and entrapment technology of active carriers which are essential in many engineering and medicine applications.

  18. Application of active quenching of second generation wire for current limiting

    SciTech Connect

    Solovyov, Vyacheslav F.; Li, Qiang

    2015-10-19

    Superconducting fault current limiters (SFCL's) are increasingly implemented in the power grid as a protection of substation equipment from fault currents. Resistive SFCL's are compact and light, however they are passively triggered and thus may not be sufficiently sensitive to respond to faults in the distribution grid. Here, we explore the prospect of adding an active management feature to a traditional resistive SFCL. A flexible radio-frequency coil, which is an integral part of the switching structure, acts as a triggering device. We show that the application of a short, 10 ms, burst of ac magnetic field during the fault triggers a uniform quench of the wire and significantly reduces the reaction time of the wire at low currents. The ac field burst generates a high density of normal zones, which merge into a continuous resistive region at a rate much faster than that of sparse normal zones created by the transport current alone.

  19. Application of active quenching of second generation wire for current limiting

    DOE PAGES

    Solovyov, Vyacheslav F.; Li, Qiang

    2015-10-19

    Superconducting fault current limiters (SFCL's) are increasingly implemented in the power grid as a protection of substation equipment from fault currents. Resistive SFCL's are compact and light, however they are passively triggered and thus may not be sufficiently sensitive to respond to faults in the distribution grid. Here, we explore the prospect of adding an active management feature to a traditional resistive SFCL. A flexible radio-frequency coil, which is an integral part of the switching structure, acts as a triggering device. We show that the application of a short, 10 ms, burst of ac magnetic field during the fault triggersmore » a uniform quench of the wire and significantly reduces the reaction time of the wire at low currents. The ac field burst generates a high density of normal zones, which merge into a continuous resistive region at a rate much faster than that of sparse normal zones created by the transport current alone.« less

  20. Application of active quenching of second generation wire for current limiting

    NASA Astrophysics Data System (ADS)

    Solovyov, Vyacheslav F.; Li, Qiang

    2015-12-01

    Superconducting fault current limiters (SFCLs) are increasingly implemented in the power grid as a protection of substation equipment from fault currents. Resistive SFCLs are compact and light, however they are passively triggered and thus may not be sufficiently sensitive to respond to faults in the distribution grid. Here, we explore the prospect of adding an active management feature to a traditional resistive SFCL. A flexible radio-frequency coil, which is an integral part of the switching structure, acts as a triggering device. We show that the application of a short, 10 ms, burst of ac magnetic field during the fault triggers a uniform quench of the wire and significantly reduces the reaction time of the wire at low currents. The ac field burst generates a high density of normal zones, which merge into a continuous resistive region at a rate much faster than that of sparse normal zones created by the transport current alone.

  1. Acoustic waves in the atmosphere and ground generated by volcanic activity

    SciTech Connect

    Ichihara, Mie; Lyons, John; Oikawa, Jun; Takeo, Minoru

    2012-09-04

    This paper reports an interesting sequence of harmonic tremor observed in the 2011 eruption of Shinmoe-dake volcano, southern Japan. The main eruptive activity started with ashcloud forming explosive eruptions, followed by lava effusion. Harmonic tremor was transmitted into the ground and observed as seismic waves at the last stage of the effusive eruption. The tremor observed at this stage had unclear and fluctuating harmonic modes. In the atmosphere, on the other hand, many impulsive acoustic waves indicating small surface explosions were observed. When the effusion stopped and the erupted lava began explosive degassing, harmonic tremor started to be transmitted also to the atmosphere and observed as acoustic waves. Then the harmonic modes became clearer and more stable. This sequence of harmonic tremor is interpreted as a process in which volcanic degassing generates an open connection between the volcanic conduit and the atmosphere. In order to test this hypothesis, a laboratory experiment was performed and the essential features were successfully reproduced.

  2. Acoustic waves in the atmosphere and ground generated by volcanic activity

    NASA Astrophysics Data System (ADS)

    Ichihara, Mie; Lyons, John; Oikawa, Jun; Takeo, Minoru

    2012-09-01

    This paper reports an interesting sequence of harmonic tremor observed in the 2011 eruption of Shinmoe-dake volcano, southern Japan. The main eruptive activity started with ashcloud forming explosive eruptions, followed by lava effusion. Harmonic tremor was transmitted into the ground and observed as seismic waves at the last stage of the effusive eruption. The tremor observed at this stage had unclear and fluctuating harmonic modes. In the atmosphere, on the other hand, many impulsive acoustic waves indicating small surface explosions were observed. When the effusion stopped and the erupted lava began explosive degassing, harmonic tremor started to be transmitted also to the atmosphere and observed as acoustic waves. Then the harmonic modes became clearer and more stable. This sequence of harmonic tremor is interpreted as a process in which volcanic degassing generates an open connection between the volcanic conduit and the atmosphere. In order to test this hypothesis, a laboratory experiment was performed and the essential features were successfully reproduced.

  3. Optics At The Arctic Circle, An Example Of Application-Oriented Research Generating New Industrial Activities

    NASA Astrophysics Data System (ADS)

    Lammasniemi, Jorma; Myllyla, Risto; Hannula, Tapio

    1989-04-01

    This paper discusses research/industry interaction in application-oriented research groups specializing in the development of optoelectronic instruments and measurement methods. The research groups are working in Oulu, a city in Northern Finland, in an industrial environment consisting originally of pulp and paper industries together with metalworking and engineering industries. These established industrial areas are active in adopting new technologies for automation and process renewal. Furthermore, new emerging businesses are being generated through pilot installations and new product ideas created by research groups. The technologies considered are optical and infrared process analyzers, semiconductor laser-based dimension measurements and optoelectronic hybrid module fabrication. Examples of new products and enterprises employing these technologies are given. Additional skills and education especially in miniature optics and related constructions, are considered important for the future.

  4. generation of picosecond pulses in solid-state lasers using new active media

    SciTech Connect

    Lisitsyn, V.N.; Matrosov, V.N.; Pestryakov, E.V.; Trunov, V.I.

    1986-07-01

    Results are reported of investigations aimed at generating nanosecond radiation pulses in solid-state lasers using new active media having broad gain lines. Passive mode locking is accomplished for the first time in a BeLa:Nd/sup 3/ laser at a wavelength 1.354 microm, and in a YAG:Nd/sup 3/ laser on a 1.32-microm transition. The free lasing and mode-locking regimes were investigated in an alexandrite (BeA1/sub 2/O/sub 4/:Cr/sup 3/) laser in the 0.72-0.78-microm range and in a synchronously pumped laser on F/sub 2//sup -/ centers in LiF in the 1.12-1.24-microm region. The features of nonlinear perception of IR radiation by the eye, using a developed picosecond laser on F/sub 2//sup -/ centers, are investigated for the first time.

  5. Generation of femtosecond optical vortices by molecular modulation in a Raman-active crystal.

    PubMed

    Zhi, Miaochan; Wang, Kai; Hua, Xia; Schuessler, Hans; Strohaber, James; Sokolov, Alexei V

    2013-11-18

    We have generated multi-color optical vortices in a Raman-active crystal PbWO4 using two-color Fourier-transform limited femtosecond laser pulses. This setup overcomes some of the limitation of our previous research by allowing for the production of subcycle femtosecond optical vortices without the need for compensating for added chirp. In addition, the use of an OPA allows for greater flexibility in exciting different Raman modes. We verified the topological charges using two different methods. These diagnostic experiments verify not only theoretically predicted OAM algebra but demonstrated instabilities in high-order OVs. We have also studied factors which affect the high-order vortex sidebands such as the diameter and intensity of the input beams.

  6. Demand generation activities and modern contraceptive use in urban areas of four countries: a longitudinal evaluation.

    PubMed

    Speizer, Ilene S; Corroon, Meghan; Calhoun, Lisa; Lance, Peter; Montana, Livia; Nanda, Priya; Guilkey, David

    2014-11-06

    Family planning is crucial for preventing unintended pregnancies and for improving maternal and child health and well-being. In urban areas where there are large inequities in family planning use, particularly among the urban poor, programs are needed to increase access to and use of contraception among those most in need. This paper presents the midterm evaluation findings of the Urban Reproductive Health Initiative (Urban RH Initiative) programs, funded by the Bill & Melinda Gates Foundation, that are being implemented in 4 countries: India (Uttar Pradesh), Kenya, Nigeria, and Senegal. Between 2010 and 2013, the Measurement, Learning & Evaluation (MLE) project collected baseline and 2-year longitudinal follow-up data from women in target study cities to examine the role of demand generation activities undertaken as part of the Urban RH Initiative programs. Evaluation results demonstrate that, in each country where it was measured, outreach by community health or family planning workers as well as local radio programs were significantly associated with increased use of modern contraceptive methods. In addition, in India and Nigeria, television programs had a significant effect on modern contraceptive use, and in Kenya and Nigeria, the program slogans and materials that were blanketed across the cities (eg, leaflets/brochures distributed at health clinics and the program logo placed on all forms of materials, from market umbrellas to health facility signs and television programs) were also significantly associated with modern method use. Our results show that targeted, multilevel demand generation activities can make an important contribution to increasing modern contraceptive use in urban areas and could impact Millennium Development Goals for improved maternal and child health and access to reproductive health for all.

  7. Demand generation activities and modern contraceptive use in urban areas of four countries: a longitudinal evaluation

    PubMed Central

    Speizer, Ilene S; Corroon, Meghan; Calhoun, Lisa; Lance, Peter; Montana, Livia; Nanda, Priya; Guilkey, David

    2014-01-01

    ABSTRACT Family planning is crucial for preventing unintended pregnancies and for improving maternal and child health and well-being. In urban areas where there are large inequities in family planning use, particularly among the urban poor, programs are needed to increase access to and use of contraception among those most in need. This paper presents the midterm evaluation findings of the Urban Reproductive Health Initiative (Urban RH Initiative) programs, funded by the Bill & Melinda Gates Foundation, that are being implemented in 4 countries: India (Uttar Pradesh), Kenya, Nigeria, and Senegal. Between 2010 and 2013, the Measurement, Learning & Evaluation (MLE) project collected baseline and 2-year longitudinal follow-up data from women in target study cities to examine the role of demand generation activities undertaken as part of the Urban RH Initiative programs. Evaluation results demonstrate that, in each country where it was measured, outreach by community health or family planning workers as well as local radio programs were significantly associated with increased use of modern contraceptive methods. In addition, in India and Nigeria, television programs had a significant effect on modern contraceptive use, and in Kenya and Nigeria, the program slogans and materials that were blanketed across the cities (eg, leaflets/brochures distributed at health clinics and the program logo placed on all forms of materials, from market umbrellas to health facility signs and television programs) were also significantly associated with modern method use. Our results show that targeted, multilevel demand generation activities can make an important contribution to increasing modern contraceptive use in urban areas and could impact Millennium Development Goals for improved maternal and child health and access to reproductive health for all. PMID:25611476

  8. Properties of rhythmic activity generated by the isolated spinal cord of the neonatal mouse.

    PubMed

    Whelan, P; Bonnot, A; O'Donovan, M J

    2000-12-01

    We examined the ability of the isolated lumbosacral spinal cord of the neonatal mouse (P0-7) to generate rhythmic motor activity under several different conditions. In the absence of electrical or pharmacological stimulation, we recorded several patterns of spontaneous ventral root depolarization and discharge. Spontaneous, alternating discharge between contralateral ventral roots could occur two to three times over a 10-min interval. We also observed other patterns, including left-right synchrony and rhythmic activity restricted to one side of the cord. Trains of stimuli delivered to the lumbar/coccygeal dorsal roots or the sural nerve reliably evoked episodes of rhythmic activity. During these evoked episodes, rhythmic ventral root discharges could occur on one side of the cord or could alternate from side to side. Bath application of a combination of N-methyl-D,L-aspartate (NMA), serotonin, and dopamine produced rhythmic activity that could last for several hours. Under these conditions, the discharge recorded from the left and right L(1)-L(3) ventral roots alternated. In the L(4)-L(5) segments, the discharge had two peaks in each cycle, coincident with discharge of the ipsilateral and contralateral L(1)-L(3) roots. The L(6) ventral root discharge alternated with that recorded from the ipsilateral L(1)-L(3) roots. We established that the drug-induced rhythm was locomotor-like by recording an alternating pattern of discharge between ipsilateral flexor and extensor hindlimb muscle nerves. In addition, by recording simultaneously from ventral roots and muscle nerves, we established that ankle flexor discharge was in phase with ipsilateral L(1)/L(2) ventral root discharge, while extensor discharge was in phase with ipsilateral L(6) ventral root discharge. Rhythmic patterns of ventral root discharge were preserved following mid-sagittal section of the spinal cord, demonstrating that reciprocal inhibitory connections between the left and right sides of the cord are

  9. PPARδ agonism inhibits skeletal muscle PDC activity, mitochondrial ATP production and force generation during prolonged contraction

    PubMed Central

    Constantin-Teodosiu, Dumitru; Baker, David J; Constantin, Despina; Greenhaff, Paul L

    2009-01-01

    We have recently shown that PPARδ agonism, used clinically to treat insulin resistance, increases fat oxidation and up-regulates mitochondrial PDK4 mRNA and protein expression in resting skeletal muscle. We hypothesized that PDK4 up-regulation, which inhibits pyruvate dehydrogenase complex (PDC)-dependent carbohydrate (CHO) oxidation, would negatively affect muscle function during sustained contraction where the demand on CHO is markedly increased. Three groups of eight male Wistar rats each received either vehicle or a PPARδ agonist (GW610742X) at two doses (5 and 100 mg (kg body mass (bm))−1 orally for 6 days. On the seventh day, the gastrocnemius–soleus–plantaris muscle group was isolated and snap frozen, or underwent 30 min of electrically evoked submaximal intensity isometric contraction using a perfused hindlimb model. During contraction, the rate of muscle PDC activation was significantly lower at 100 mg (kg bm)−1 compared with control (P < 0.01). Furthermore, the rates of muscle PCr hydrolysis and lactate accumulation were significantly increased at 100 mg (kg bm)−1 compared with control, reflecting lower mitochondrial ATP generation. Muscle tension development during contraction was significantly lower at 100 mg (kg bm)−1 compared with control (25%; P < 0.05). The present data demonstrate that PPARδ agonism inhibits muscle CHO oxidation at the level of PDC during prolonged contraction, and is paralleled by the activation of anaerobic metabolism, which collectively impair contractile function. PMID:19001043

  10. Generation 9 polyamidoamine dendrimer encapsulated platinum nanoparticle mimics catalase size, shape, and catalytic activity.

    PubMed

    Wang, Xinyu; Zhang, Yincong; Li, Tianfu; Tian, Wende; Zhang, Qiang; Cheng, Yiyun

    2013-04-30

    Poly(amidoamine) (PAMAM) encapsulated platinum nanoparticles were synthesized and used as catalase mimics. Acetylated generation 9 (Ac-G9) PAMAM dendrimer with a molecular size around 10 nm was used as a template to synthesize platinum nanoparticles. The feeding molar ratio of Pt(4+) and Ac-G9 is 2048, and the synthesized platinum nanoparticle (Ac-G9/Pt NP) has an average size of 3.3 nm. Ac-G9/Pt NP has a similar molecular size and globular shape with catalase (~11 nm). The catalytic activity of Ac-G9/Pt NP on the decomposition of H2O2 is approaching that of catalase at 37 °C. Ac-G9/Pt NP shows differential response to the changes of pH and temperature compared with catalase, which can be explained by different catalytic mechanisms of Ac-G9/Pt NP and catalase. Ac-G9/Pt NP also shows horseradish peroxidase activity and is able to scavenge free radicals such as di(phenyl)-(2,4,6-trinitrophenyl)iminoazanium (DPPH). Furthermore, Ac-G9/Pt NP shows excellent biocompatibility on different cell lines and can down-regulate H2O2-induced intracellular reactive oxygen species (ROS) in these cells. These results suggest that dendrimers are promising mimics of proteins with different sizes and Ac-G9/Pt NP can be used as an alternative candidate of catalase to decrease oxidation stress in cells.

  11. Recent Data Generation Activities at the Atomic and Molecular Data Unit of the IAEA

    SciTech Connect

    Clark, R.E.H.; Humbert, D.

    2005-05-27

    The main data generation mechanism of the Atomic and Molecular (A+M) Data Unit of the IAEA is the Co-ordinated Research Project (CRP). The International Fusion Research Council Subcommittee on Atomic and Molecular Data for Fusion recommends topics for new CRPs to be initiated by the A+M Unit. A typical CRP has a lifetime of three to five years. At the start of the CRP a Research Co-ordination Meeting (RCM) is held with the purpose of formulating a detailed work plan. At later RCMs progress on these work plans is reported and the studies debated and expanded. At the conclusion of the CRP the results are compiled in a volume of the journal Atomic and Plasma-Material Interaction Data for Fusion. Numerical results are also added to the electronic database as appropriate. Normally the Unit has three to four active CRPs, and also holds Technical Meetings and invites individual Consultants to IAEA Headquarters, Vienna for specific tasks. Such activities can result in providing advice on a particular topic, on data for a particular process, or a new capability to be made widely available. Recently, consultants to the Unit have provided extensive additions to the Unit databases, as well as interfaces to run several calculational tools through the Internet. Specific examples will be presented.

  12. Inflammatory cytokines presented from polymer matrices differentially generate and activate DCs in situ

    PubMed Central

    Ali, Omar A.; Tayalia, Prakriti; Shvartsman, Dmitry; Lewin, Sarah; Mooney, David J.

    2014-01-01

    During infection, inflammatory cytokines mobilize and activate dendritic cells (DCs), which are essential for efficacious T cell priming and immune responses that clear the infection. Here we designed macroporous poly(lactide-co-glycolide) (PLG) matrices to release the inflammatory cytokines GM-CSF, Flt3L and CCL20, in order to mimic infection-induced DC recruitment. We then tested the ability of these infection mimics to function as cancer vaccines via induction of specific, anti-tumor T cell responses. All vaccine systems tested were able to confer specific anti-tumor T cell responses and longterm survival in a therapeutic, B16-F10 melanoma model. However, GM-CSF and Flt3L vaccines resulted in similar survival rates, and outperformed CCL20 loaded scaffolds, even though they had differential effects on DC recruitment and generation. GM-CSF signaling was identified as the most potent chemotactic factor for conventional DCs and significantly enhanced surface expression of MHC(II) and CD86(+), which are utilized for priming T cell immunity. In contrast, Flt3L vaccines led to greater numbers of plasmacytoid DCs (pDCs), correlating with increased levels of T cell priming cytokines that amplify T cell responses. These results demonstrate that 3D polymer matrices modified to present inflammatory cytokines may be utilized to effectively mobilize and activate different DC subsets in vivo for immunotherapy. PMID:24688455

  13. NASA's Advanced Propulsion Technology Activities for Third Generation Fully Reusable Launch Vehicle Applications

    NASA Technical Reports Server (NTRS)

    Hueter, Uwe

    2000-01-01

    NASA's Office of Aeronautics and Space Transportation Technology (OASTT) established the following three major goals, referred to as "The Three Pillars for Success": Global Civil Aviation, Revolutionary Technology Leaps, and Access to Space. The Advanced Space Transportation Program Office (ASTP) at the NASA's Marshall Space Flight Center in Huntsville, Ala. focuses on future space transportation technologies under the "Access to Space" pillar. The Propulsion Projects within ASTP under the investment area of Spaceliner100, focus on the earth-to-orbit (ETO) third generation reusable launch vehicle technologies. The goals of Spaceliner 100 is to reduce cost by a factor of 100 and improve safety by a factor of 10,000 over current conditions. The ETO Propulsion Projects in ASTP, are actively developing combination/combined-cycle propulsion technologies that utilized airbreathing propulsion during a major portion of the trajectory. System integration, components, materials and advanced rocket technologies are also being pursued. Over the last several years, one of the main thrusts has been to develop rocket-based combined cycle (RBCC) technologies. The focus has been on conducting ground tests of several engine designs to establish the RBCC flowpaths performance. Flowpath testing of three different RBCC engine designs is progressing. Additionally, vehicle system studies are being conducted to assess potential operational space access vehicles utilizing combined-cycle propulsion systems. The design, manufacturing, and ground testing of a scale flight-type engine are planned. The first flight demonstration of an airbreathing combined cycle propulsion system is envisioned around 2005. The paper will describe the advanced propulsion technologies that are being being developed under the ETO activities in the ASTP program. Progress, findings, and future activities for the propulsion technologies will be discussed.

  14. Novel keto-phospholipids are generated by monocytes and macrophages, detected in cystic fibrosis, and activate peroxisome proliferator-activated receptor-γ.

    PubMed

    Hammond, Victoria J; Morgan, Alwena H; Lauder, Sarah; Thomas, Christopher P; Brown, Sarah; Freeman, Bruce A; Lloyd, Clare M; Davies, Jane; Bush, Andrew; Levonen, Anna-Liisa; Kansanen, Emilia; Villacorta, Luis; Chen, Y Eugene; Porter, Ned; Garcia-Diaz, Yoel M; Schopfer, Francisco J; O'Donnell, Valerie B

    2012-12-07

    12/15-Lipoxygenases (LOXs) in monocytes and macrophages generate novel phospholipid-esterified eicosanoids. Here, we report the generation of two additional families of related lipids comprising 15-ketoeicosatetraenoic acid (KETE) attached to four phosphatidylethanolamines (PEs). The lipids are generated basally by 15-LOX in IL-4-stimulated monocytes, are elevated on calcium mobilization, and are detected at increased levels in bronchoalveolar lavage fluid from cystic fibrosis patients (3.6 ng/ml of lavage). Murine peritoneal macrophages generate 12-KETE-PEs, which are absent in 12/15-LOX-deficient mice. Inhibition of 15-prostaglandin dehydrogenase prevents their formation from exogenous 15-hydroxyeicosatetraenoic acid-PE in human monocytes. Both human and murine cells also generated analogous hydroperoxyeicosatetraenoic acid-PEs. The electrophilic reactivity of KETE-PEs is shown by their Michael addition to glutathione and cysteine. Lastly, both 15-hydroxyeicosatetraenoic acid-PE and 15-KETE-PE activated peroxisome proliferator-activated receptor-γ reporter activity in macrophages in a dose-dependent manner. In summary, we demonstrate novel peroxisome proliferator-activated receptor-γ-activating oxidized phospholipids generated enzymatically by LOX and 15-prostaglandin dehydrogenase in primary monocytic cells and in a human Th2-related lung disease. The lipids are a new family of bioactive mediators from the 12/15-LOX pathway that may contribute to its known anti-inflammatory actions in vivo.

  15. Methylglyoxal-induced modification of arginine residues decreases the activity of NADPH-generating enzymes.

    PubMed

    Morgan, Philip E; Sheahan, Pamela J; Pattison, David I; Davies, Michael J

    2013-08-01

    Inadequate control of plasma and cellular glucose and ketone levels in diabetes is associated with increased generation of reactive aldehydes, including methylglyoxal (MGO). These aldehydes react with protein side chains to form advanced glycation end-products (AGEs). Arg residues are particularly susceptible to MGO glycation and are essential for binding NADP(+) in several enzymes that generate NADPH, a coenzyme for many critical metabolic and antioxidant enzymes. In most animal cells, NADPH is produced predominantly by glucose-6-phosphate dehydrogenase (G6PD) in the oxidative phase of the pentose phosphate pathway and, to a lesser extent, by isocitrate dehydrogenase (IDH) and malic enzyme (ME). In this study, the activities of isolated G6PD, IDH, and ME were inhibited by MGO (0-2.5mM, 2-3h, 37°C), in a dose- and time-dependent manner, with G6PD and IDH more sensitive to modification than ME. Significant inhibition of these two enzymes occurred with MGO levels ≥500μM. Incubation with radiolabeled MGO (0-500µM, 0-3h, 37°C) demonstrated dose- and time-dependent adduction to G6PD and IDH. HPLC analysis provided evidence for AGE formation and particularly the hydroimidazolones MG-H1 and MG-H2 from Arg residues, with corresponding loss of parent Arg residues. Peptide mass mapping studies confirmed hydroimidazolone formation on multiple peptides in G6PD and IDH, including those critical for NADP(+) binding, and substrate binding, in the case of IDH. These results suggest that modification of NADPH-producing enzymes by reactive aldehydes may result in alterations to the cellular redox environment, potentially predisposing cells to further damage by oxidants and reactive aldehydes.

  16. Next-generation active and passive heatsink design for diode lasers

    NASA Astrophysics Data System (ADS)

    Srinivasan, Raman; Miller, Robert; Kuppuswamy, Kiran; Nguyen, Touyen; Hu, Yongdan; Li, Hanxuan; Brown, Dennis; Towe, Terry; Crum, Trevor; Morris, Bob; Wolak, Ed; Harrison, Jim

    2007-02-01

    Successful thermal and stress management of edge-emitting GaAs-based diode lasers is key to their performance and reliability in high-power operation. Complementary to advanced epitaxial structures and die-fabrication processes, next-generation heatsink designs are required to meet the requirements of emerging applications. In this paper, we detail the development of both active and passive heatsinks designed to match the coefficient of thermal expansion (CTE) of the laser die. These CTE-matched heatsinks also offer low thermal resistance, compatibility with AuSn bonding and improved manufacturability. Early data representing the performance of high-power devices on the new heatsinks are included in the presentation. Among the designs are a water-cooled, mini-channel heatsink with a CTE of 6.8 ppm/°C (near to the nominal 6.5 ppm/°C CTE of GaAs) and a thermal resistance of 0.43 °C/W (assuming a 27%-fill-factor diode-laser bar with a cavity length of 2 mm). The water flow in the heatsink is isolated from the electrical potential, eliminating the possibility of electrolytic corrosion. An additional feature of the integrated design is the reduction in required assembly steps. Our next-generation, passive, CTE-matched heatsink employs a novel design to achieve a reduction of 16% in thermal resistance (compared to the predecessor commercial product). CTE's can be engineered to fall in the range of 6.2-7.2 ppm/°C on the bar mounting surface. Comparisons between simulated performance and experimental data (both in CW and long-pulse operation) will be presented for several new heat-sink designs.

  17. G-EVER Activities and the Next-generation Volcanic Hazard Assessment System

    NASA Astrophysics Data System (ADS)

    Takarada, S.

    2013-12-01

    The Asia-Pacific Region Global Earthquake and Volcanic Eruption Risk Management (G-EVER) is a consortium of Asia-Pacific geohazard research institutes that was established in 2012. G-EVER aims to formulate strategies to reduce the risks of disasters worldwide caused by the occurrence of earthquakes, tsunamis and volcanic eruptions. G-EVER is working on enhancing collaboration, sharing of resources, and making information on the risks of earthquakes and volcanic eruptions freely available and understandable. The 1st G-EVER International Symposium was held in Tsukuba, Japan in March 11, 2013. The 2nd Symposium is scheduled in Sendai, Tohoku Japan, in Oct. 19-20, 2013. Currently, 4 working groups were proposed in the G-EVER Consortium. The next-generation volcano hazard assessment WG is developing a useful system for volcanic eruption prediction, risk assessment, and evacuation at various eruption stages. The assessment system is based on volcanic eruption history datasets, volcanic eruption database, and numerical simulations. Volcanic eruption histories including precursor phenomena leading to major eruptions of active volcanoes are very important for future prediction of volcanic eruptions. A high quality volcanic eruption database, which contains compilations of eruption dates, volumes, and types, is important for the next-generation volcano hazard assessment system. Proposing international standards on how to estimate the volume of volcanic products is important to make a high quality volcanic eruption database. Spatial distribution database of volcanic products (e.g. tephra and pyroclastic flow distributions), encoded into a GIS based database is necessary for more precise area and volume estimation and risk assessments. The volcanic eruption database is developed based on past eruption results, which only represents a subset of possible future scenarios. Therefore, numerical simulations with controlled parameters are needed for more precise volcanic eruption

  18. Analysis of the contribution of FTO, NPC1, ENPP1, NEGR1, GNPDA2 and MC4R genes to obesity in Mexican children

    PubMed Central

    2013-01-01

    Background Recent genome wide association studies (GWAS) and previous positional linkage studies have identified more than 50 single nucleotide polymorphisms (SNPs) associated with obesity, mostly in Europeans. We aimed to assess the contribution of some of these SNPs to obesity risk and to the variation of related metabolic traits, in Mexican children. Methods The association of six European obesity-related SNPs in or near FTO, NPC1, ENPP1, NEGR1, GNPDA2 and MC4R genes with risk of obesity was tested in 1,463 school-aged Mexican children (Ncases = 514; Ncontrols = 949). We also assessed effects of these SNPs on the variation of body mass index (BMI), fasting serum insulin levels, fasting plasma glucose levels, total cholesterol and triglyceride levels, in a subset of 1,171 nonobese Mexican children. Results We found a significant effect of GNPDA2 rs10938397 on risk of obesity (odds ratio [OR] = 1.30; P = 1.34 × 10-3). Furthermore, we found nominal associations between obesity risk or BMI variation and the following SNPs: ENPP1 rs7754561, MC4R rs17782313 and NEGR1 rs2815752. Importantly, the at-risk alleles of both MC4R rs17782313 and NPC1 rs1805081 showed significant effect on increased fasting glucose levels (β = 0.36 mmol/L; P = 1.47 × 10-3) and decreased fasting serum insulin levels (β = −0.10 μU/mL; P = 1.21 × 10-3), respectively. Conclusion Our present results suggest that some obesity-associated SNPs previously reported in Europeans also associate with risk of obesity, or metabolic quantitative traits, in Mexican children. Importantly, we found new associations between MC4R and fasting glucose levels, and between NPC1 and fasting insulin levels. PMID:23375129

  19. Lysophosphatidic acid induces reactive oxygen species generation by activating protein kinase C in PC-3 human prostate cancer cells

    SciTech Connect

    Lin, Chu-Cheng; Lin, Chuan-En; Lin, Yueh-Chien; Ju, Tsai-Kai; Huang, Yuan-Li; Lee, Ming-Shyue; Chen, Jiun-Hong; Lee, Hsinyu

    2013-11-01

    Highlights: •LPA induces ROS generation through LPA{sub 1} and LPA{sub 3}. •LPA induces ROS generation by activating PLC. •PKCζ mediates LPA-induced ROS generation. -- Abstract: Prostate cancer is one of the most frequently diagnosed cancers in males, and PC-3 is a cell model popularly used for investigating the behavior of late stage prostate cancer. Lysophosphatidic acid (LPA) is a lysophospholipid that mediates multiple behaviors in cancer cells, such as proliferation, migration and adhesion. We have previously demonstrated that LPA enhances vascular endothelial growth factor (VEGF)-C expression in PC-3 cells by activating the generation of reactive oxygen species (ROS), which is known to be an important mediator in cancer progression. Using flow cytometry, we showed that LPA triggers ROS generation within 10 min and that the generated ROS can be suppressed by pretreatment with the NADPH oxidase (Nox) inhibitor diphenylene iodonium. In addition, transfection with LPA{sub 1} and LPA{sub 3} siRNA efficiently blocked LPA-induced ROS production, suggesting that both receptors are involved in this pathway. Using specific inhibitors and siRNA, phospholipase C (PLC) and protein kinase C (PKC) were also suggested to participate in LPA-induced ROS generation. Overall, we demonstrated that LPA induces ROS generation in PC-3 prostate cancer cells and this is mediated through the PLC/PKC/Nox pathway.

  20. Recovery of strontium activity from a strontium-82/rubidium-82 generator

    DOEpatents

    Taylor, Wayne A.; Phillips, Dennis R.; Sosnowski, Kenneth M.

    1999-10-12

    Strontium-82 is recovered from spent strontium-82/rubidium-82 generators to provide a source of strontium-82 for additional strontium-82/rubidium-82 generators. The process involves stripping of the strontium-82 from used strontium-82/rubidium-82 generators followed by purification of the strontium-82 material to remove additional metal contaminants to desired levels.

  1. The protein level and transcription activity of activating transcription factor 1 is regulated by prolyl isomerase Pin1 in nasopharyngeal carcinoma progression.

    PubMed

    Huang, Guo-Liang; Liao, Dan; Chen, Hua; Lu, Yan; Chen, Liyong; Li, Huahui; Li, Binbin; Liu, Weilong; Ye, Caiguo; Li, Tong; Zhu, Zhu; Wang, Jian; Uchida, Takafumi; Zou, Ying; Dong, Zigang; He, Zhiwei

    2016-12-29

    The function of activating transcription factor 1 (ATF1) and the mechanism about why ATF1 was over-phosphorylated in nasopharyngeal carcinoma (NPC) progression is completely undiscovered. In this study, a series of experiments both in vitro and in vivo were used to characterize a promotive function of ATF1 in NPC tumorigenesis and identify prolyl isomerase Pin1 as a novel regulator of ATF1 at post-transcription. First, we found that overexpression of ATF1 promoted colony formation in NPC. However, the high protein level of ATF1 in NPC was not resulted from high mRNA level. Then, a direct interaction between Pin1 and ATF1 at Thr184 was demonstrated using mammalian two-hybrid assay and coimmunoprecipitation. Cycloheximide (CHX) treatment indicated Pin1 stabilized the expression of ATF1 at post-transcription level. We confirmed that Pin1 upregulated ATF1 transcriptional activity of Bcl-2 using luciferase reporter assay, quantitative RT-PCR and western blot. Furthermore, the newly identified phosphorylation of ATF1 at Thr184 was suggested to have an important role in ATF1 function of transcription and tumor promotion. Finally, high expression of Pin1 in NPC tissue was found to be positively correlated with ATF1. The ATF1 promoted NPC tumorigenesis was regulated by Pin1 both in vitro and in vivo. All these findings clearly state that Pin1 is a novel regulator of ATF1 at Thr184 and thereby enhances ATF1 transcription activity and tumorigenesis promotive function in NPC.

  2. The protein level and transcription activity of activating transcription factor 1 is regulated by prolyl isomerase Pin1 in nasopharyngeal carcinoma progression

    PubMed Central

    Huang, Guo-Liang; Liao, Dan; Chen, Hua; Lu, Yan; Chen, Liyong; Li, Huahui; Li, Binbin; Liu, Weilong; Ye, Caiguo; Li, Tong; Zhu, Zhu; Wang, Jian; Uchida, Takafumi; Zou, Ying; Dong, Zigang; He, Zhiwei

    2016-01-01

    The function of activating transcription factor 1 (ATF1) and the mechanism about why ATF1 was over-phosphorylated in nasopharyngeal carcinoma (NPC) progression is completely undiscovered. In this study, a series of experiments both in vitro and in vivo were used to characterize a promotive function of ATF1 in NPC tumorigenesis and identify prolyl isomerase Pin1 as a novel regulator of ATF1 at post-transcription. First, we found that overexpression of ATF1 promoted colony formation in NPC. However, the high protein level of ATF1 in NPC was not resulted from high mRNA level. Then, a direct interaction between Pin1 and ATF1 at Thr184 was demonstrated using mammalian two-hybrid assay and coimmunoprecipitation. Cycloheximide (CHX) treatment indicated Pin1 stabilized the expression of ATF1 at post-transcription level. We confirmed that Pin1 upregulated ATF1 transcriptional activity of Bcl-2 using luciferase reporter assay, quantitative RT-PCR and western blot. Furthermore, the newly identified phosphorylation of ATF1 at Thr184 was suggested to have an important role in ATF1 function of transcription and tumor promotion. Finally, high expression of Pin1 in NPC tissue was found to be positively correlated with ATF1. The ATF1 promoted NPC tumorigenesis was regulated by Pin1 both in vitro and in vivo. All these findings clearly state that Pin1 is a novel regulator of ATF1 at Thr184 and thereby enhances ATF1 transcription activity and tumorigenesis promotive function in NPC. PMID:28032861

  3. Derivation of rat embryonic stem cells and generation of protease-activated receptor-2 knockout rats.

    PubMed

    Yamamoto, Satoshi; Nakata, Mitsugu; Sasada, Reiko; Ooshima, Yuki; Yano, Takashi; Shinozawa, Tadahiro; Tsukimi, Yasuhiro; Takeyama, Michiyasu; Matsumoto, Yoshio; Hashimoto, Tadatoshi

    2012-08-01

    One of the remarkable achievements in knockout (KO) rat production reported during the period 2008-2010 is the derivation of authentic embryonic stem (ES) cells from rat blastocysts using a novel culture medium containing glycogen synthase kinase 3 and mitogen-activated protein kinase kinase inhibitors (2i medium). Here, we report gene-targeting technology via homologous recombination in rat ES cells, demonstrating its use through production of a protease-activated receptor-2 gene (Par-2) KO rat. We began by generating germline-competent ES cells from Dark Agouti rats using 2i medium. These ES cells, which differentiate into cardiomyocytes in vitro, can produce chimeras with high ES cell contribution when injected into blastocysts. We then introduced a targeting vector with a neomycin-resistant gene driven by the CAG promoter to disrupt Par-2. After a 7-day drug selection, 489 neomycin-resistant colonies were obtained. Following screening by polymerase chain reaction (PCR) genotyping and quantitative PCR analysis, we confirmed three homologous recombinant clones, resulting in chimeras that transmitted the Par-2 targeted allele to offspring. Par-2 KO rats showed a loss of Par-2 messenger RNA expression in their stomach cells and a lack of PAR-2 mediated smooth muscle relaxation in the aorta as indicated by pharmacological testing. Compared with mice, rats offer many advantages in biomedical research, including a larger body size; consequently, they are widely used in scientific investigation. Thus, the establishment of a gene-targeting technology using rat ES cells will be a valuable tool in human disease model production and drug discovery.

  4. A dosimetry study of deuterium-deuterium neutron generator-based in vivo neutron activation analysis

    NASA Astrophysics Data System (ADS)

    Sowers, Daniel A.

    A neutron irradiation cavity for in vivo Neutron Activation Analysis (IVNAA) to detect manganese, aluminum, and other potentially toxic elements in human hand bone has been designed and its dosimetric specifications measured. The neutron source is a customized deuterium-deuterium neutron generator which produces neutrons at 2.45 MeV by the fusion reaction 2H(d, n)3He at a calculated flux of 7 x 108 +/-30% s-1. A moderator/reflector/shielding (5 cm high density polyethylene (HDPE), 5.3 cm graphite & 5.7 cm borated HDPE) assembly has been designed and built to maximize the thermal neutron flux inside the hand irradiation cavity and to reduce the extremity dose and effective dose to the human subject. Lead sheets are used to attenuate bremsstrahlung x rays and activation gammas. A Monte Carlo simulation (MCNP6) was used to model the system and calculate extremity dose. The extremity dose was measured with neutron and photon sensitive film badges and Fuji electronic pocket dosimeter (EPD). The neutron ambient dose outside the shielding was measured by Fuji NSN3, and photon dose by a Bicron MicroREM scintillator. Neutron extremity dose was calculated to be 32.3 mSv using MCNP6 simulations given a 10 min IVNAA measurement of manganese. Measurements by EPD and film badge indicate hand dose to be 31.7 +/- 0.8 mSv for neutron and 4.2 +/- 0.2 mSv for photon for 10 mins; whole body effective dose was calculated conservatively to be 0.052 mSv. Experimental values closely match values obtained from MCNP6 simulations. These are acceptable doses to apply the technology for a manganese toxicity study in a human population.

  5. A Dosimetry Study of Deuterium-Deuterium Neutron Generator-based In Vivo Neutron Activation Analysis.

    PubMed

    Sowers, Daniel; Liu, Yingzi; Mostafaei, Farshad; Blake, Scott; Nie, Linda H

    2015-12-01

    A neutron irradiation cavity for in vivo neutron activation analysis (IVNAA) to detect manganese, aluminum, and other potentially toxic elements in human hand bone has been designed and its dosimetric specifications measured. The neutron source is a customized deuterium-deuterium neutron generator that produces neutrons at 2.45 MeV by the fusion reaction 2H(d, n)3He at a calculated flux of 7 × 10(8) ± 30% s(-1). A moderator/reflector/shielding [5 cm high density polyethylene (HDPE), 5.3 cm graphite and 5.7 cm borated (HDPE)] assembly has been designed and built to maximize the thermal neutron flux inside the hand irradiation cavity and to reduce the extremity dose and effective dose to the human subject. Lead sheets are used to attenuate bremsstrahlung x rays and activation gammas. A Monte Carlo simulation (MCNP6) was used to model the system and calculate extremity dose. The extremity dose was measured with neutron and photon sensitive film badges and Fuji electronic pocket dosimeters (EPD). The neutron ambient dose outside the shielding was measured by Fuji NSN3, and the photon dose was measured by a Bicron MicroREM scintillator. Neutron extremity dose was calculated to be 32.3 mSv using MCNP6 simulations given a 10-min IVNAA measurement of manganese. Measurements by EPD and film badge indicate hand dose to be 31.7 ± 0.8 mSv for neutrons and 4.2 ± 0.2 mSv for photons for 10 min; whole body effective dose was calculated conservatively to be 0.052 mSv. Experimental values closely match values obtained from MCNP6 simulations. These are acceptable doses to apply the technology for a manganese toxicity study in a human population.

  6. Are Older Adults Physically Active Enough – A Matter of Assessment Method? The Generation 100 Study

    PubMed Central

    Zisko, Nina; Ingebrigtsen, Jan Erik; Wisløff, Ulrik; Stensvold, Dorthe

    2016-01-01

    Introduction Physical activity (PA) is beneficial for general health. As a result, adults around the world are recommended to undertake regular PA of either absolute or relative intensity. Traditionally, adherence to PA recommendation is assessed by accelerometers that record absolute intensity thresholds. Since ageing often results in a decrease in cardiorespiratory fitness (CRF), older adults (aged > 65 years) might be more susceptible to not meeting the PA recommendation when measured in absolute terms. The aim of the present study was to compare the adherence to the PA recommendation using both absolute and relative thresholds. Additionally, we aimed to report the reference values for overall PA in a large sample of Norwegian older adults. Methods PA was assessed for 7 days using the Actigraph GT3X+ accelerometer in 1219 older adults (624 females) aged 70–77 years. Overall PA was measured as counts per minute (CPM) and steps. Absolute and relative moderate-to-vigorous PA (MVPA) thresholds were applied to quantify adherence to PA recommendation. The relative MVPA thresholds were developed specifically for the Generation 100 population sample. CRF was directly measured as peak oxygen uptake (VO2peak). Results Proportions meeting PA recommendation were 29% and 71% when utilizing absolute and relative MVPA, respectively. More females met the relative PA recommendation compared to males. Overall PA was higher among the youngest age group. Older adults with medium- and high levels of CRF were more physically active, compared to those with the lowest levels of CRF. Conclusion This is the first study to compare adherence to PA recommendation, using absolute and relative intensity thresholds among older adults. The present study clearly illustrates the consequences of using different methodological approaches to surveillance of PA across age, gender and CRF in a population of older adults. PMID:27893785

  7. Social studies of volcanology: knowledge generation and expert advice on active volcanoes

    NASA Astrophysics Data System (ADS)

    Donovan, Amy; Oppenheimer, Clive; Bravo, Michael

    2012-04-01

    This paper examines the philosophy and evolution of volcanological science in recent years, particularly in relation to the growth of volcanic hazard and risk science. It uses the lens of Science and Technology Studies to examine the ways in which knowledge generation is controlled and directed by social forces, particularly during eruptions, which constitute landmarks in the development of new technologies and models. It also presents data from a survey of volcanologists carried out during late 2008 and early 2009. These data concern the felt purpose of the science according to the volcanologists who participated and their impressions of the most important eruptions in historical time. It demonstrates that volcanologists are motivated both by the academic science environment and by a social concern for managing the impact of volcanic hazards on populations. Also discussed are the eruptions that have most influenced the discipline and the role of scientists in policymaking on active volcanoes. Expertise in volcanology can become the primary driver of public policy very suddenly when a volcano erupts, placing immense pressure on volcanologists. In response, the epistemological foundations of volcanology are on the move, with an increasing volume of research into risk assessment and management. This requires new, integrated methodologies for knowledge collection that transcend scientific disciplinary boundaries.

  8. Ameloblasts require active RhoA to generate normal dental enamel.

    PubMed

    Xue, Hui; Li, Yong; Everett, Eric T; Ryan, Kathleen; Peng, Li; Porecha, Rakhee; Yan, Yan; Lucchese, Anna M; Kuehl, Melissa A; Pugach, Megan K; Bouchard, Jessica; Gibson, Carolyn W

    2013-08-01

    RhoA plays a fundamental role in regulation of the actin cytoskeleton, intercellular attachment, and cell proliferation. During amelogenesis, ameloblasts (which produce the enamel proteins) undergo dramatic cytoskeletal changes and the RhoA protein level is up-regulated. Transgenic mice were generated that express a dominant-negative RhoA transgene in ameloblasts using amelogenin gene-regulatory sequences. Transgenic and wild-type (WT) molar tooth germs were incubated with sodium fluoride (NaF) or sodium chloride (NaCl) in organ culture. Filamentous actin (F-actin) stained with phalloidin was elevated significantly in WT ameloblasts treated with NaF compared with WT ameloblasts treated with NaCl or with transgenic ameloblasts treated with NaF, thereby confirming a block in the RhoA/Rho-associated protein kinase (ROCK) pathway in the transgenic mice. Little difference in quantitative fluorescence (an estimation of fluorosis) was observed between WT and transgenic incisors from mice provided with drinking water containing NaF. We subsequently found reduced transgene expression in incisors compared with molars. Transgenic molar teeth had reduced amelogenin, E-cadherin, and Ki67 compared with WT molar teeth. Hypoplastic enamel in transgenic mice correlates with reduced expression of the enamel protein, amelogenin, and E-cadherin and cell proliferation are regulated by RhoA in other tissues. Together these findings reveal deficits in molar ameloblast function when RhoA activity is inhibited.

  9. Ameloblasts require active RhoA to generate normal dental enamel

    PubMed Central

    Li, Yong; Xue, Hui; Everett, Eric T.; Ryan, Kathleen; Peng, Li; Porecha, Rakhee; Yan, Yan; Lucchese, Anna M.; Kuehl, Melissa A.; Pugach, Megan K.; Bouchard, Jessica; Gibson, Carolyn W.

    2013-01-01

    RhoA plays a fundamental role in regulation of the actin cytoskeleton, intercellular attachment and cell proliferation. During amelogenesis, ameloblasts which produce the enamel proteins undergo dramatic cytoskeletal changes and RhoA protein level is upregulated. Transgenic mice were generated that express a dominant-negative RhoA transgene in ameloblasts using amelogenin gene regulatory sequences. Transgenic and WT molar tooth germs were incubated with NaF or NaCl in organ culture. F-actin stained with phalloidin was elevated significantly in WT ameloblasts treated with NaF compared to WT ameloblasts treated with NaCl or compared to transgenic ameloblasts treated with NaF, thereby confirming a block in the RhoA/ROCK pathway in the transgenic mice. Little difference in quantitative fluorescence (estimation of fluorosis) was observed between WT and transgenic incisors from mice provided NaF in their drinking water. We subsequently found reduced transgene expression in incisors compared to molars. Transgenic molar teeth had reduced amelogenin, E-cadherin and Ki67 compared to WT. Hypoplastic enamel in transgenic mice correlates with reduced expression of the enamel protein amelogenin, and E-cadherin and cell proliferation are regulated by RhoA in other tissues. Together these findings reveal deficits in molar ameloblast function when RhoA activity is inhibited. PMID:23841780

  10. Experiments on the motion of gas bubbles in turbulence generated by an active grid

    NASA Astrophysics Data System (ADS)

    Poorte, R. E. G.; Biesheuvel, A.

    2002-06-01

    The random motion of nearly spherical bubbles in the turbulent flow behind a grid is studied experimentally. In quiescent water these bubbles rise at high Reynolds number. The turbulence is generated by an active grid of the design of Makita (1991), and can have turbulence Reynolds number R[lambda] of up to 200. Minor changes in the geometry of the grid and in its mode of operation improves the isotropy of the turbulence, compared with that reported by Makita (1991) and Mydlarski & Warhaft (1996). The trajectory of each bubble is measured with high spatial and temporal resolution with a specially developed technique that makes use of a position-sensitive detector. Bubble statistics such as the mean rise velocity and the root-mean-square velocity fluctuations are obtained by ensemble averaging over many identical bubbles. The resulting bubble mean rise velocity is significantly reduced (up to 35%) compared with the quiescent conditions. The vertical bubble velocity fluctuations are found to be non-Gaussian, whereas the horizontal displacements are Gaussian for all times. The diffusivity of bubbles is considerably less than that of fluid particles. These findings are qualitatively consistent with results obtained through theoretical analysis and numerical simulations by Spelt & Biesheuvel (1997).

  11. The new generation drug candidate molecules: Spectral, electrochemical, DNA-binding and anticancer activity properties

    NASA Astrophysics Data System (ADS)

    Gölcü, Ayşegül; Muslu, Harun; Kılıçaslan, Derya; Çeşme, Mustafa; Eren, Özge; Ataş, Fatma; Demirtaş, İbrahim

    2016-09-01

    The new generation drug candidate molecules [Cu(5-Fu)2Cl2H2O] (NGDCM1) and [Zn(5-Fu)2(CH3COO)2] (NGDCM2) were obtained from the reaction of copper(II) and zinc(II) salts with the anticancer drug 5-fluoracil (5-Fu). These compounds have been characterized by spectroscopic and analytical techniques. Thermal behavior of the compounds were also investigated. The electrochemical properties of the compounds have been investigated by cyclic voltammetry (CV) using glassy carbon electrode. The biological activity of the NGDCM1 and NGDCM2 has been evaluated by examining their ability to bind to fish sperm double strand DNA (FSdsDNA) with UV spectroscopy. UV studies of the interaction of the 5-Fu and metal derivatives with FSdsDNA have shown that these compounds can bind to FSdsDNA. The binding constants of the compounds with FSdsDNA have also been calculated. Thermal decomposition of the compounds lead to the formation of CuO and ZnO as final products. The effect of proliferation 5-Fu, NGDCM1 and NGDCM2 were examined on the HeLa cells using real-time cell analyzer with three different concentrations.

  12. Development of new generation reduced activation ferritic-martensitic steels for advanced fusion reactors

    NASA Astrophysics Data System (ADS)

    Tan, L.; Snead, L. L.; Katoh, Y.

    2016-09-01

    International development of reduced activation ferritic-martensitic (RAFM) steels has focused on 9 wt percentage Cr, which primarily contain M23C6 (M = Cr-rich) and small amounts of MX (M = Ta/V, X = C/N) precipitates, not adequate to maintain strength and creep resistance above ∼500 °C. To enable applications at higher temperatures for better thermal efficiency of fusion reactors, computational alloy thermodynamics coupled with strength modeling have been employed to explore a new generation RAFM steels. The new alloys are designed to significantly increase the amount of MX nanoprecipitates, which are manufacturable through standard and scalable industrial steelmaking methods. Preliminary experimental results of the developed new alloys demonstrated noticeably increased amount of MX, favoring significantly improved strength, creep resistance, and Charpy impact toughness as compared to current RAFM steels. The strength and creep resistance were comparable or approaching to the lower bound of, but impact toughness was noticeably superior to 9-20Cr oxide dispersion-strengthened ferritic alloys.

  13. Entropy generation in a parallel-plate active magnetic regenerator with insulator layers

    NASA Astrophysics Data System (ADS)

    Mugica Guerrero, Ibai; Poncet, Sébastien; Bouchard, Jonathan

    2017-02-01

    This paper proposes a feasible solution to diminish conduction losses in active magnetic regenerators. Higher performances of these machines are linked to a lower thermal conductivity of the Magneto-Caloric Material (MCM) in the streamwise direction. The concept presented here involves the insertion of insulator layers along the length of a parallel-plate magnetic regenerator in order to reduce the heat conduction within the MCM. This idea is investigated by means of a 1D numerical model. This model solves not only the energy equations for the fluid and solid domains but also the magnetic circuit that conforms the experimental setup of reference. In conclusion, the addition of insulator layers within the MCM increases the temperature span, cooling load, and coefficient of performance by a combination of lower heat conduction losses and an increment of the global Magneto-Caloric Effect. The generated entropy by solid conduction, fluid convection, and conduction and viscous losses are calculated to help understand the implications of introducing insulator layers in magnetic regenerators. Finally, the optimal number of insulator layers is studied.

  14. Spatiotemporal imaging of cortical activation during verb generation and picture naming.

    PubMed

    Edwards, Erik; Nagarajan, Srikantan S; Dalal, Sarang S; Canolty, Ryan T; Kirsch, Heidi E; Barbaro, Nicholas M; Knight, Robert T

    2010-03-01

    One hundred and fifty years of neurolinguistic research has identified the key structures in the human brain that support language. However, neither the classic neuropsychological approaches introduced by Broca (1861) and Wernicke (1874), nor modern neuroimaging employing PET and fMRI has been able to delineate the temporal flow of language processing in the human brain. We recorded the electrocorticogram (ECoG) from indwelling electrodes over left hemisphere language cortices during two common language tasks, verb generation and picture naming. We observed that the very high frequencies of the ECoG (high-gamma, 70-160 Hz) track language processing with spatial and temporal precision. Serial progression of activations is seen at a larger timescale, showing distinct stages of perception, semantic association/selection, and speech production. Within the areas supporting each of these larger processing stages, parallel (or "incremental") processing is observed. In addition to the traditional posterior vs. anterior localization for speech perception vs. production, we provide novel evidence for the role of premotor cortex in speech perception and of Wernicke's and surrounding cortex in speech production. The data are discussed with regards to current leading models of speech perception and production, and a "dual ventral stream" hybrid of leading speech perception models is given.

  15. Development of new generation reduced activation ferritic-martenstic steels for advanced fusion reactors

    SciTech Connect

    Tan, Lizhen; Snead, Lance Lewis; Katoh, Yutai

    2016-05-26

    International development of reduced activation ferritic-martensitic (RAFM) steels has focused on 9 wt percentage Cr, which primarily contain M23C6 (M = Cr-rich) and small amounts of MX (M = Ta/V, X = C/N) precipitates, not adequate to maintain strength and creep resistance above ~500 °C. To enable applications at higher temperatures for better thermal efficiency of fusion reactors, computational alloy thermodynamics coupled with strength modeling have been employed to explore a new generation RAFM steels. The new alloys are designed to significantly increase the amount of MX nanoprecipitates, which are manufacturable through standard and scalable industrial steelmaking methods. Preliminary experimental results of the developed new alloys demonstrated noticeably increased amount of MX, favoring significantly improved strength, creep resistance, and Charpy impact toughness as compared to current RAFM steels. Furthermore, the strength and creep resistance were comparable or approaching to the lower bound of, but impact toughness was noticeably superior to 9–20Cr oxide dispersion-strengthened ferritic alloys.

  16. Development of new generation reduced activation ferritic-martenstic steels for advanced fusion reactors

    DOE PAGES

    Tan, Lizhen; Snead, Lance Lewis; Katoh, Yutai

    2016-05-26

    International development of reduced activation ferritic-martensitic (RAFM) steels has focused on 9 wt percentage Cr, which primarily contain M23C6 (M = Cr-rich) and small amounts of MX (M = Ta/V, X = C/N) precipitates, not adequate to maintain strength and creep resistance above ~500 °C. To enable applications at higher temperatures for better thermal efficiency of fusion reactors, computational alloy thermodynamics coupled with strength modeling have been employed to explore a new generation RAFM steels. The new alloys are designed to significantly increase the amount of MX nanoprecipitates, which are manufacturable through standard and scalable industrial steelmaking methods. Preliminary experimentalmore » results of the developed new alloys demonstrated noticeably increased amount of MX, favoring significantly improved strength, creep resistance, and Charpy impact toughness as compared to current RAFM steels. Furthermore, the strength and creep resistance were comparable or approaching to the lower bound of, but impact toughness was noticeably superior to 9–20Cr oxide dispersion-strengthened ferritic alloys.« less

  17. Meprin Metalloproteases Generate Biologically Active Soluble Interleukin-6 Receptor to Induce Trans-Signaling

    PubMed Central

    Arnold, Philipp; Boll, Inga; Rothaug, Michelle; Schumacher, Neele; Schmidt, Frederike; Wichert, Rielana; Schneppenheim, Janna; Lokau, Juliane; Pickhinke, Ute; Koudelka, Tomas; Tholey, Andreas; Rabe, Björn; Scheller, Jürgen; Lucius, Ralph; Garbers, Christoph; Rose-John, Stefan; Becker-Pauly, Christoph

    2017-01-01

    Soluble Interleukin-6 receptor (sIL-6R) mediated trans-signaling is an important pro-inflammatory stimulus associated with pathological conditions, such as arthritis, neurodegeneration and inflammatory bowel disease. The sIL-6R is generated proteolytically from its membrane bound form and A Disintegrin And Metalloprotease (ADAM) 10 and 17 were shown to perform ectodomain shedding of the receptor in vitro and in vivo. However, under certain conditions not all sIL-6R could be assigned to ADAM10/17 activity. Here, we demonstrate that the IL-6R is a shedding substrate of soluble meprin α and membrane bound meprin β, resulting in bioactive sIL-6R that is capable of inducing IL-6 trans-signaling. We determined cleavage within the N-terminal part of the IL-6R stalk region, distinct from the cleavage site reported for ADAM10/17. Interestingly, meprin β can be shed from the cell surface by ADAM10/17 and the observation that soluble meprin β is not capable of shedding the IL-6R suggests a regulatory mechanism towards trans-signaling. Additionally, we observed a significant negative correlation of meprin β expression and IL-6R levels on human granulocytes, providing evidence for in vivo function of this proteolytic interaction. PMID:28276471

  18. C1 Metabolism Inhibition and Nitrogen Deprivation Trigger Triacylglycerol Accumulation in Arabidopsis thaliana Cell Cultures and Highlight a Role of NPC in Phosphatidylcholine-to-Triacylglycerol Pathway

    PubMed Central

    Meï, Coline E.; Cussac, Mathilde; Haslam, Richard P.; Beaudoin, Frédéric; Wong, Yung-Sing; Maréchal, Eric; Rébeillé, Fabrice

    2017-01-01

    Triacylglycerol (TAG) accumulation often occurs in growth limiting conditions such as nutrient deprivations. We analyzed and compared the lipid contents of Arabidopsis cells grown under two conditions that inhibited growth as a way to study interactions between membrane and storage lipids. In order to inhibit C1 metabolism, the first condition utilized methotrexate (MTX), a drug that inhibits methyl transfer reactions and potentially reduces Pi-choline synthesis, the polar head of phosphatidylcholine (PC). MTX-treated cells displayed a 10- to 15-fold increase in TAG compared to that found in control cells. This corresponded to a net increase of lipids as the total amount of membrane glycerolipids was minimally affected. Under this condition, PC homeostasis appeared tightly regulated and not strictly dependent on the rate of Pi-choline synthesis. The second condition we investigated involved nitrogen deprivation. Here, we observed a 40-fold increase of TAG. In these cells, the overall lipid content remained unchanged, but membrane lipids decreased by a factor of two suggesting a reduction of the membrane network and a rerouting of membrane lipids to storage lipids. Under all conditions, fatty acid (FA) analyses showed that the FA composition of TAG was comparable to that in PC, but different from that in acyl-CoA, suggesting that TAG accumulation involved PC-derived DAG moieties. In agreement, analyses by qPCR of genes coding for TAG synthesis showed a strong increase of non-specific phospholipase C (NPC) expressions, and experiments using labeled (fluorescent) PC indicated higher rates of PC-to-TAG conversion under both situations. These results highlight a role for NPC in plant cell oil production. PMID:28101097

  19. Active vortex generator deployed on demand by size independent actuation of shape memory alloy wires integrated in fiber reinforced polymers

    NASA Astrophysics Data System (ADS)

    Hübler, M.; Nissle, S.; Gurka, M.; Wassenaar, J.

    2016-04-01

    Static vortex generators (VGs) are installed on different aircraft types. They generate vortices and interfuse the slow boundary layer with the fast moving air above. Due to this energizing, a flow separation of the boundary layer can be suppressed at high angles of attack. However the VGs cause a permanently increased drag over the whole flight cycle reducing the cruise efficiency. This drawback is currently limiting the use of VGs. New active VGs, deployed only on demand at low speed, can help to overcome this contradiction. Active hybrid structures, combining the actuation of shape memory alloys (SMA) with fiber reinforced polymers (FRP) on the materials level, provide an actuation principle with high lightweight potential and minimum space requirements. Being one of the first applications of active hybrid structures from SMA and FRP, these active vortex generators help to demonstrate the advantages of this new technology. A new design approach and experimental results of active VGs are presented based on the application of unique design tools and advanced manufacturing approaches for these active hybrid structures. The experimental investigation of the actuation focuses on the deflection potential and the dynamic response. Benchmark performance data such as a weight of 1.5g and a maximum thickness of only 1.8mm per vortex generator finally ensure a simple integration in the wing structure.

  20. Synaptic generation of an intracellular retrograde signal requires activation of the tyrosine kinase and mitogen-activated protein kinase signaling cascades in Aplysia.

    PubMed

    Stough, Shara; Kopec, Ashley M; Carew, Thomas J

    2015-11-01

    Cellular changes underlying memory formation can be generated in an activity-dependent manner at specific synapses. Thus an important question concerns the mechanisms by which synaptic signals communicate with the cell body to mediate these cellular changes. A monosynaptic circuit that is enhanced by sensitization in Aplysia is well-suited to study this question because three different subcellular compartments: (i) the sensorimotor SN-MN synapses, (ii) the SN projections to MNs via axonal connections, (iii) the SN cell bodies, can all be manipulated and studied independently. Here, we report that activity-dependent (AD) training in either the entire SN-MN circuit or in only the synaptic compartment, activates MAPK in a temporally and spatially specific pattern. Specifically, we find (i) MAPK activation is first transiently generated at SN-MN synapses during training, (ii) immediately after training MAPK is transiently activated in SN-MN axonal connections and persistently activated in SN cell bodies, and finally, (iii) MAPK is activated in SN cell bodies and SN-MN synapses 1h after training. These data suggest that there is an intracellularly transported retrograde signal generated at the synapse which is later responsible for delayed MAPK activation at SN somata. Finally, we find that this retrograde signal requires activation of tyrosine kinase (TK) and MEK signaling cascades at the synapses.

  1. Induction of ROS generation by fluconazole in Candida glabrata: activation of antioxidant enzymes and oxidative DNA damage.

    PubMed

    Mahl, Camila Donato; Behling, Camile Saul; Hackenhaar, Fernanda S; de Carvalho e Silva, Mélany Natuane; Putti, Jordana; Salomon, Tiago B; Alves, Sydney Hartz; Fuentefria, Alexandre; Benfato, Mara S

    2015-07-01

    In this study, we assessed the generation of reactive oxygen species (ROS) induced by subinhibitory concentration of fluconazole in susceptible and resistant Candida glabrata strains at stationary growth phase and measured their oxidative responses parameters: glutathione peroxidase (GPx), superoxide dismutase (SOD), glutathione-S-transferase (GST), consumption of hydrogen peroxide, and total glutathione, as well as oxidative damage in lipids, proteins, and DNA. Data showed that fluconazole increased generation of ROS and GPx and SOD enzymatic activity in treated cells; however, these enzymatic activities did not differ between resistant and susceptible strains. Susceptible strains exhibited higher GST activity than resistant, and when susceptible cells were treated with fluconazole, GST activity decreased. Fluconazole treatment cause oxidative damage only in DNA. There are a possible participation of ROS, as organic peroxides and O2(•-), in antifungal mechanism of fluconazole, which results in higher GPx and SOD enzymatic activities and oxidative DNA damage in C. glabrata.

  2. Coupling template nanocasting and self-activation for fabrication of nanoporous carbon

    PubMed Central

    Kong, Lingjun; Liu, Mingxiang; Diao, Zenghui; Chen, Diyun; Chang, Xiangyang; Xiong, Ya

    2016-01-01

    Hierarchical nanoporous carbon (NPC) with great surface area and developed pore size distribution has been intently concerned. Herein, we report a facile method coupling template nanocasting and self-activation to fabricate nanoporous carbon with continuous micro, meso and macro pores, in which CaCO3 acted as template and activation reagent while the flour was the carbon precursor. Effects of mass ratio of CaCO3 to flour and carbonized temperature on the pore structures of NPC were investigated by nitrogen adsorption-desorption isotherms and SEM analysis. Another kind of carbon was prepared by directly mixed powder CaCO3 with flour carbonized at 800 °C (NPC-p) to comparatively investigate the pore fabricating mechanism. Results shown that carbonized at 800 °C was favorable to fabricate the continuous macro, meso and micro pores. The resulted NPC in a mass ratio of 1 to 2 had the considerable SBET and VT of 575.4 m2/g and 0.704 cm3/g, respectively. Only surface activation was observed for NPC-p. Nanocasting of the powder CaCO3 contributed to fabricate macropores and the CO2 activation contributed to meso- and micropores. Coupling activation and nanocasting effect due to the decomposition of CaCO3 template into CO2 and CaO was ascribed to synthesize the nanoporous carbon. PMID:27901109

  3. Coupling template nanocasting and self-activation for fabrication of nanoporous carbon

    NASA Astrophysics Data System (ADS)

    Kong, Lingjun; Liu, Mingxiang; Diao, Zenghui; Chen, Diyun; Chang, Xiangyang; Xiong, Ya

    2016-11-01

    Hierarchical nanoporous carbon (NPC) with great surface area and developed pore size distribution has been intently concerned. Herein, we report a facile method coupling template nanocasting and self-activation to fabricate nanoporous carbon with continuous micro, meso and macro pores, in which CaCO3 acted as template and activation reagent while the flour was the carbon precursor. Effects of mass ratio of CaCO3 to flour and carbonized temperature on the pore structures of NPC were investigated by nitrogen adsorption-desorption isotherms and SEM analysis. Another kind of carbon was prepared by directly mixed powder CaCO3 with flour carbonized at 800 °C (NPC-p) to comparatively investigate the pore fabricating mechanism. Results shown that carbonized at 800 °C was favorable to fabricate the continuous macro, meso and micro pores. The resulted NPC in a mass ratio of 1 to 2 had the considerable SBET and VT of 575.4 m2/g and 0.704 cm3/g, respectively. Only surface activation was observed for NPC-p. Nanocasting of the powder CaCO3 contributed to fabricate macropores and the CO2 activation contributed to meso- and micropores. Coupling activation and nanocasting effect due to the decomposition of CaCO3 template into CO2 and CaO was ascribed to synthesize the nanoporous carbon.

  4. Octopamine-containing (OC) interneurons enhance central pattern generator activity in sucrose-induced feeding in the snail Lymnaea.

    PubMed

    Vehovszky, Agnes; Szabó, Henriette; Elliott, Christopher J H

    2004-10-01

    In the pond snail Lymnaea stagnalis octopamine-containing (OC) interneurons trigger and reconfigure the feeding pattern in isolated CNS by excitation of the central pattern generator. In semi-intact (lip-mouth-CNS) preparations, this central pattern generator is activated by chemosensory inputs. We now test if sucrose application to the lips activates the OC neurons independently of the rest of the feeding central pattern generator, or if the OC interneuron is activated by inputs from the feeding network. In 66% of experiments, sucrose stimulated feeding rhythms and OC interneurons received regular synaptic inputs. Only rarely (14%) did the OC interneuron fire action potentials, proving that firing of OC interneurons is not necessary for the sucrose-induced feeding. Prestimulation of OC neurons increased the intensity and duration of the feeding rhythm evoked by subsequent sucrose presentations. One micromolar octopamine in the CNS bath mimicked the effect of OC interneuron stimulation, enhancing the feeding response when sucrose is applied to the lips. We conclude that the modulatory OC neurons are not independently excited by chemosensory inputs to the lips, but rather from the buccal central pattern generator network. However, when OC neurons fire, they release modulatory octopamine, which provides a positive feedback to the network to enhance the sucrose-activated central pattern generator rhythm.

  5. Actual operation and regulatory activities on steam generator replacement in Japan

    SciTech Connect

    Saeki, Hitoshi

    1997-02-01

    This paper summarizes the operating reactors in Japan, and the status of the steam generators in these plants. It reviews plans for replacement of existing steam generators, and then goes into more detail on the planning and regulatory steps which must be addressed in the process of accomplishing this maintenance. The paper also reviews the typical steps involved in the process of removal and replacement of steam generators.

  6. Identification of serum component involved in generation of neo-lectin with agglutinating and phenoloxidase activities in human serum.

    PubMed

    Manikandan, Beulaja; Ramar, Manikandan; Munusamy, Arumugam

    2014-01-01

    Human serum albumin (HSA) was identified as the component involved in generation of neo-lectin molecules with both lectin and phenoloxidase activities. Pronase treated HSA was able to agglutinate hen RBC and oxidize hydroquinone. Sodium dodecyl sulphate (SDS) treated HSA agglutinated both hen and sheep RBC as well as oxidized dopamine. The hemagglutinating activities of pronase/SDS treated HSA observed against hen RBC were dosimetric. The oxidation of pronase/SDS treated HSA with hydroquinone/dopamine, respectively, was inhibitable by inhibitors of phenoloxidase, namely, phenylthiourea and tropolone. Very low concentrations of HSA could generate these humoral neo-lectin molecules.

  7. Association Between User-Generated Commuting Data and Population-Representative Active Commuting Surveillance Data - Four Cities, 2014-2015.

    PubMed

    Whitfield, Geoffrey P; Ussery, Emily N; Riordan, Brian; Wendel, Arthur M

    2016-09-16

    Creating environments that support all types of physical activity, including active transportation, is a public health priority (1). Public health surveillance that identifies the locations where community members walk and bicycle (i.e., engage in active transportation) can inform such efforts. Traditional population-representative active transportation surveillance incurs a considerable time lag between data collection and dissemination, and often lacks geographic specificity (2). Conversely, user-generated active transportation data from Global Positioning System (GPS)-based activity tracking devices and mobile applications can provide near real-time information, but might be subject to self-selection bias among users. CDC analyzed the association between GPS-based commuting data from a company that allows tracking of activity with a mobile application (Strava, Inc., San Francisco, California) and population-representative commuting data from the U.S. Census Bureau's American Community Survey (ACS) (3) for four U.S. cities. The level of analysis was the Census block group. The number of GPS-tracked commuters in Strava was associated with the number of ACS active commuters (Spearman's rho = 0.60), suggesting block groups were ranked similarly based on these distinct but related measurements. The correlation was higher in high population density areas. User-generated active transportation data might complement traditional surveillance systems by providing near real-time, location-specific information on where active transportation occurs.

  8. High-fidelity MCNP modeling of a D-T neutron generator for active interrogation of special nuclear material

    NASA Astrophysics Data System (ADS)

    Katalenich, Jeff; Flaska, Marek; Pozzi, Sara A.; Hartman, Michael R.

    2011-10-01

    Fast and robust methods for interrogation of special nuclear material (SNM) are of interest to many agencies and institutions in the United States. It is well known that passive interrogation methods are typically sufficient for plutonium identification because of a relatively high neutron production rate from 240Pu [1]. On the other hand, identification of shielded uranium requires active methods using neutron or photon sources [2]. Deuterium-deuterium (2.45 MeV) and deuterium-tritium (14.1 MeV) neutron-generator sources have been previously tested and proven to be relatively reliable instruments for active interrogation of nuclear materials [3,4]. In addition, the newest generators of this type are small enough for applications requiring portable interrogation systems. Active interrogation techniques using high-energy neutrons are being investigated as a method to detect hidden SNM in shielded containers [4,5]. Due to the thickness of some containers, penetrating radiation such as high-energy neutrons can provide a potential means of probing shielded SNM. In an effort to develop the capability to assess the signal seen from various forms of shielded nuclear materials, the University of Michigan Neutron Science Laboratory's D-T neutron generator and its shielding were accurately modeled in MCNP. The generator, while operating at nominal power, produces approximately 1×10 10 neutrons/s, a source intensity which requires a large amount of shielding to minimize the dose rates around the generator. For this reason, the existing shielding completely encompasses the generator and does not include beam ports. Therefore, several MCNP simulations were performed to estimate the yield of uncollided 14.1-MeV neutrons from the generator for active interrogation experiments. Beam port diameters of 5, 10, 15, 20, and 25 cm were modeled to assess the resulting neutron fluxes. The neutron flux outside the beam ports was estimated to be approximately 2×10 4 n/cm 2 s.

  9. Nitric Oxide (NO) Generation from Heme/Copper Assembly Mediated Nitrite Reductase Activity

    PubMed Central

    Hematian, Shabnam; Siegler, Maxime A.

    2014-01-01

    Nitric oxide (NO) as a cellular signaling molecule and vasodilator regulates a range of physiological and pathological processes. Nitrite (NO2−) is recycled in vivo to generate nitric oxide, particularly in physiologic hypoxia and ischemia. The cytochrome c oxidase (CcO) binuclear hemea3/CuB active site is one entity known to be responsible for cellular nitrite conversion to nitric oxide. We recently reported that a partially reduced heme/Cu assembly reduces nitrite ion, producing NO; the heme serves as the reductant and cupric ion provides a Lewis Acid interaction with nitrite, facilitating nitrite (N−O) bond cleavage (Hematian et al., J Am Chem Soc 134:18912–18915, 2012). To further investigate this nitrite reductase (NIR) chemistry, copper(II)-nitrito complexes with tri-and tetra-dentate ligands were used in this study, where either O,O'-bidentate or O-unidentate modes of nitrite binding to the cupric center are present. To study the role of the reducing ability of the ferrous heme center, two different tetraarylporphyrinate-iron(II) complexes, one with electron donating para-methoxy peripheral substituents, (TMPP)FeII, and the other with electron withdrawing 2,6-difluorophenyl substituents, (F8)FeII, were employed. The results show that differing nitrite coordination modes to copper(II) ion leads to varying kinetic behavior. Here, also, the ferrous heme is in all cases the source of the reducing equivalent required to take nitrite to nitric oxide, but the reduction ability of the heme center does not play a key role in the observed overall reaction rate. Based on our observations, reaction mechanisms are proposed and discussed in terms of heme/Cu heterobinuclear structures. PMID:24430198

  10. Nitric oxide generation from heme/copper assembly mediated nitrite reductase activity.

    PubMed

    Hematian, Shabnam; Siegler, Maxime A; Karlin, Kenneth D

    2014-06-01

    Nitric oxide (NO) as a cellular signaling molecule and vasodilator regulates a range of physiological and pathological processes. Nitrite (NO2 (-)) is recycled in vivo to generate nitric oxide, particularly in physiologic hypoxia and ischemia. The cytochrome c oxidase binuclear heme a 3/CuB active site is one entity known to be responsible for conversion of cellular nitrite to nitric oxide. We recently reported that a partially reduced heme/copper assembly reduces nitrite ion, producing nitric oxide; the heme serves as the reductant and the cupric ion provides a Lewis acid interaction with nitrite, facilitating nitrite (N-O) bond cleavage (Hematian et al., J. Am. Chem. Soc. 134:18912-18915, 2012). To further investigate this nitrite reductase chemistry, copper(II)-nitrito complexes with tridentate and tetradentate ligands were used in this study, where either O,O'-bidentate or O-unidentate modes of nitrite binding to the cupric center are present. To study the role of the reducing ability of the ferrous heme center, two different tetraarylporphyrinate-iron(II) complexes, one with electron-donating para-methoxy peripheral substituents and the other with electron-withdrawing 2,6-difluorophenyl substituents, were used. The results show that differing modes of nitrite coordination to the copper(II) ion lead to differing kinetic behavior. Here, also, the ferrous heme is in all cases the source of the reducing equivalent required to convert nitrite to nitric oxide, but the reduction ability of the heme center does not play a key role in the observed overall reaction rate. On the basis of our observations, reaction mechanisms are proposed and discussed in terms of heme/copper heterobinuclear structures.

  11. Cloud condensation nucleus activity comparison of dry- and wet-generated mineral dust aerosol: the significance of soluble material

    NASA Astrophysics Data System (ADS)

    Garimella, S.; Huang, Y.-w.; Seewald, J. S.; Cziczo, D. J.

    2013-11-01

    This study examines the interaction of clay mineral particles and water vapor to determine the conditions required for cloud droplet formation. Droplet formation conditions are investigated for three clay minerals: illite, sodium-rich montmorillonite, and Arizona Test Dust. Using wet and dry particle generation coupled to a differential mobility analyzer (DMA) and cloud condensation nuclei counter, the critical activation of the clay mineral particles as cloud condensation nuclei is characterized. Electron microscopy (EM) is used to determine non-sphericity in particle shape. EM is also used to determine particle surface area and account for transmission of multiply charged particles by the DMA. Single particle mass spectrometry and ion chromatography are used to investigate soluble material in wet-generated samples and demonstrate that wet and dry generation yield compositionally different particles. Activation results are analyzed in the context of both κ-Köhler theory and Frenkel, Halsey, and Hill (FHH) adsorption activation theory. This study has two main results: (1) κ-Köhler is a suitable framework, less complex than FHH theory, to describe clay mineral nucleation activity despite apparent differences in κ with respect to size. For dry-generated particles the size dependence is likely an artifact of the shape of the size distribution: there is a sharp drop-off in particle concentration at ~300 nm, and a large fraction of particles classified with a mobility diameter less than ~300 nm are actually multiply charged, resulting in a much lower critical supersaturation for droplet activation than expected. For wet-generated particles, deviation from κ-Köhler theory is likely a result of the dissolution and redistribution of soluble material. (2) Wet-generation is found to be unsuitable for simulating the lofting of fresh dry dust because it changes the size-dependent critical supersaturations by fractionating and re-partitioning soluble material.

  12. Combustion-derived flame generated ultrafine soot generates reactive oxygen species and activates Nrf2 antioxidants differently in neonatal and adult rat lungs

    PubMed Central

    2013-01-01

    Background Urban particulate matter (PM) has been epidemiologically correlated with multiple cardiopulmonary morbidities and mortalities, in sensitive populations. Children exposed to PM are more likely to develop respiratory infections and asthma. Although PM originates from natural and anthropogenic sources, vehicle exhaust rich in polycyclic aromatic hydrocarbons (PAH) can be a dominant contributor to the PM2.5 and PM0.1 fractions and has been implicated in the generation of reactive oxygen species (ROS). Objectives Current studies of ambient PM are confounded by the variable nature of PM, so we utilized a previously characterized ethylene-combusted premixed flame particles (PFP) with consistent and reproducible physiochemical properties and 1) measured the oxidative potential of PFP compared to ambient PM, 2) determined the ability of PFPs to generate oxidative stress and activate the transcription factor using in vitro and ex vivo models, and 3) we correlated these responses with antioxidant enzyme expression in vivo. Methods We compared oxidative stress response (HMOX1) and antioxidant enzyme (SOD1, SOD2, CAT, and PRDX6) expression in vivo by performing a time-course study in 7-day old neonatal and young adult rats exposed to a single 6-hour exposure to 22.4 μg/m3 PFPs. Results We showed that PFP is a potent ROS generator that induces oxidative stress and activates Nrf2. Induction of the oxidative stress responsive enzyme HMOX1 in vitro was mediated through Nrf2 activation and was variably upregulated in both ages. Furthermore, antioxidant enzyme expression had age and lung compartment variations post exposure. Of particular interest was SOD1, which had mRNA and protein upregulation in adult parenchyma, but lacked a similar response in neonates. Conclusions We conclude that PFPs are effective ROS generators, comparable to urban ambient PM2.5, that induce oxidative stress in neonatal and adult rat lungs. PFPs upregulate a select set of antioxidant enzymes in

  13. Inorganic Polyphosphates Regulate Hexokinase Activity and Reactive Oxygen Species Generation in Mitochondria of Rhipicephalus (Boophilus) microplus Embryo

    PubMed Central

    Fraga, Amanda; Moraes, Jorge; da Silva, José Roberto; Costa, Evenilton P.; Menezes, Jackson; da Silva Vaz Jr, Itabajara; Logullo, Carlos; da Fonseca, Rodrigo Nunes; Campos, Eldo

    2013-01-01

    The physiological roles of polyphosphates (poly P) recently found in arthropod mitochondria remain obscure. Here, the possible involvement of poly P with reactive oxygen species generation in mitochondria of Rhipicephalus microplus embryos was investigated. Mitochondrial hexokinase and scavenger antioxidant enzymes, such as superoxide dismutase, catalase, and glutathione reductase were assayed during embryogenesis of R. microplus. The influence of poly P3 and poly P15 were analyzed during the period of higher enzymatic activity during embryogenesis. Both poly Ps inhibited hexokinase activity by up to 90% and, interestingly, the mitochondrial membrane exopolyphosphatase activity was stimulated by the hexokinase reaction product, glucose-6-phosphate. Poly P increased hydrogen peroxide generation in mitochondria in a situation where mitochondrial hexokinase is also active. The superoxide dismutase, catalase and glutathione reductase activities were higher during embryo cellularization, at the end of embryogenesis and during embryo segmentation, respectively. All of the enzymes were stimulated by poly P3. However, superoxide dismutase was not affected by poly P15, catalase activity was stimulated only at high concentrations and glutathione reductase was the only enzyme that was stimulated in the same way by both poly Ps. Altogether, our results indicate that inorganic polyphosphate and mitochondrial membrane exopolyphosphatase regulation can be correlated with the generation of reactive oxygen species in the mitochondria of R. microplus embryos. PMID:23983617

  14. Preparation of magnetic Co/graphene sheets composites and investigation on its catalytic activity for H2 generation

    NASA Astrophysics Data System (ADS)

    Zhao, Dongcui; Nan, Zhaodong

    2016-12-01

    A cobalt (Co)/graphene sheets (GRs) composite was synthesized via a one-pot chemical method. The composite shows high saturation magnetizations (Ms), which leads it to be conveniently separated from aqueous solution by an external magnetic field. Compared to the pure Co and some references, the catalytic activity of the as-obtained composite was significantly enhanced for the generation of H2 gas by hydrolysis of NaBH4 solution. Effects of NaBH4 initial concentration, the composite and reaction temperature on the H2 generation rate were investigated. The H2 generation rate is independent with the initial NaBH4 concentration, increased with the reaction temperature increasing. The composite can be continuously used several times with about the same catalytic activity.

  15. Exploring Shifts in Middle School Learners' Modeling Activity While Generating Drawings, Animations, and Computational Simulations of Molecular Diffusion

    ERIC Educational Resources Information Center

    Wilkerson-Jerde, Michelle H.; Gravel, Brian E.; Macrander, Christopher A.

    2015-01-01

    Modeling and using technology are two practices of particular interest to K-12 science educators. These practices are inextricably linked among professionals, who engage in modeling activity with and across a variety of representational technologies. In this paper, we explore the practices of five sixth-grade girls as they generated models of…

  16. Targeting Breast Cancers Featuring Activating Mutations in PIK3CA by Generating a Lethal Dose of PIP3

    DTIC Science & Technology

    2009-02-01

    AD_________________ AWARD NUMBER: W81XWH-06-1-0341 TITLE: Targeting Breast Cancers Featuring...ORGANIZATION: Dana-Farber Cancer Institute Boston, MA 02115 REPORT DATE...2006 – 31 Jan 2009 4. TITLE AND SUBTITLE 5a. CONTRACT NUMBER Targeting Breast Cancers Featuring Activating Mutations in PIK3CA by Generating a

  17. EGCG Inhibits Proliferation, Invasiveness and Tumor Growth by Up-Regulation of Adhesion Molecules, Suppression of Gelatinases Activity, and Induction of Apoptosis in Nasopharyngeal Carcinoma Cells

    PubMed Central

    Fang, Chih-Yeu; Wu, Chung-Chun; Hsu, Hui-Yu; Chuang, Hsin-Ying; Huang, Sheng-Yen; Tsai, Ching-Hwa; Chang, Yao; Tsao, George Sai-Wah; Chen, Chi-Long; Chen, Jen-Yang

    2015-01-01

    (−)-Epigallocatechin-3-gallate (EGCG), a major green tea polyphenol, has been shown to inhibit the proliferation of a variety of tumor cells. Epidemiological studies have shown that drinking green tea can reduce the incidence of nasopharyngeal carcinoma (NPC), yet the underlying mechanism is not well understood. In this study, the inhibitory effect of EGCG was tested on a set of Epstein Barr virus-negative and -positive NPC cell lines. Treatment with EGCG inhibited the proliferation of NPC cells but did not affect the growth of a non-malignant nasopharyngeal cell line, NP460hTert. Moreover, EGCG treated cells had reduced migration and invasive properties. The expression of the cell adhesion molecules E-cadherin and β-catenin was found to be up-regulated by EGCG treatment, while the down-regulation of matrix metalloproteinases (MMP)-2 and MMP-9 were found to be mediated by suppression of extracellular signal-regulated kinase (ERK) phosphorylation and AP-1 and Sp1 transactivation. Spheroid formation by NPC cells in suspension was significantly inhibited by EGCG. Oral administration of EGCG was capable of suppressing tumor growth in xenografted mice bearing NPC tumors. Treatment with EGCG was found to elevate the expression of p53 and p21, and eventually led to apoptosis of NPC cells via caspase 3 activation. The nuclear translocation of NF-κB and β-catenin was also suppressed by EGCG treatment. These results indicate that EGCG can inhibit the proliferation and invasiveness, and induce apoptosis, of NPC cells, making it a promising agent for chemoprevention or adjuvant therapy of NPC. PMID:25625511

  18. SUMF1 mutations affecting stability and activity of formylglycine generating enzyme predict clinical outcome in multiple sulfatase deficiency.

    PubMed

    Schlotawa, Lars; Ennemann, Eva Charlotte; Radhakrishnan, Karthikeyan; Schmidt, Bernhard; Chakrapani, Anupam; Christen, Hans-Jürgen; Moser, Hugo; Steinmann, Beat; Dierks, Thomas; Gärtner, Jutta

    2011-03-01

    Multiple Sulfatase Deficiency (MSD) is caused by mutations in the sulfatase-modifying factor 1 gene encoding the formylglycine-generating enzyme (FGE). FGE post translationally activates all newly synthesized sulfatases by generating the catalytic residue formylglycine. Impaired FGE function leads to reduced sulfatase activities. Patients display combined clinical symptoms of single sulfatase deficiencies. For ten MSD patients, we determined the clinical phenotype, FGE expression, localization and stability, as well as residual FGE and sulfatase activities. A neonatal, very severe clinical phenotype resulted from a combination of two nonsense mutations leading to almost fully abrogated FGE activity, highly unstable FGE protein and nearly undetectable sulfatase activities. A late infantile mild phenotype resulted from FGE G263V leading to unstable protein but high residual FGE activity. Other missense mutations resulted in a late infantile severe phenotype because of unstable protein with low residual FGE activity. Patients with identical mutations displayed comparable clinical phenotypes. These data confirm the hypothesis that the phenotypic outcome in MSD depends on both residual FGE activity as well as protein stability. Predicting the clinical course in case of molecularly characterized mutations seems feasible, which will be helpful for genetic counseling and developing therapeutic strategies aiming at enhancement of FGE.

  19. Simulated vibrational sum frequency generation from a multilayer thin film system with two active interfaces.

    PubMed

    O'Brien, Daniel B; Massari, Aaron M

    2013-04-21

    In the field of surface-specific vibrational sum frequency generation spectroscopy (VSFG) on organic thin films, optical interferences combined with the two-interface problem presents a challenge in terms of qualitative assessment of the data and quantitative modeling. The difficulty is amplified when considering systems comprised of more than a single material thin film layer. Recently, in our lab we have developed a generalized model that describes thin film interference in interface-specific nonlinear optical spectroscopies from arbitrary multilayer systems. Here, we apply the model to simulate VSFG spectra from the simplest multilayer: a system of two thin films, one of which is an organic small molecule and the other is a dielectric layer on a semiconductor substrate system where we idealize that the organic interfaces are equally VSFG active. Specifically, we consider the molecule N,N'-dioctyl-3,4,9,10-perylenedicarboximide (PTCDI-C8) deposited on a silicon wafer with a thermally grown oxide dielectric. We present results for the four polarization experiments that sample the nonzero nonlinear susceptibility elements of macroscopically centrosymmetric materials (ssp, sps, pss, and ppp) and in two mIR frequency windows (the imide carbonyl stretches around 1680 cm(-1) and the alkyl stretches around 2900 cm(-1)) as a function of both thin film thicknesses with fixed input beam angles. We use frequency dependent refractive indices for all materials. The goal is to illustrate some of the intricacies contained in the intensity data of such systems. Of particular interest is the effect of the relative polar orientation of modes at the interfaces and the possibility of designing a system where the collected signal is exclusively attributable to a single interface. Our calculations indicate that in order to unambiguously identify the relative polar orientation one must experimentally vary an additional system parameter such as thin film thickness or input beam angle

  20. Human Recombinant Cytochrome P450 Enzymes Display Distinct Hydrogen Peroxide Generating Activities During Substrate Independent NADPH Oxidase Reactions

    PubMed Central

    Mishin, Vladimir; Heck, Diane E.; Laskin, Debra L.; Laskin, Jeffrey D.

    2014-01-01

    Microsomal enzymes generate H2O2 in the presence of NADPH. In this reaction, referred to as “oxidase” activity, H2O2 is generated directly or indirectly via the formation of superoxide anion. In the presence of redox active transition metals, H2O2 can form highly toxic hydroxyl radicals and, depending on the “oxidase” activity of individual cytochrome P450 isoenzymes, this can compromise cellular functioning and contribute to tissue injury. In the present studies, we compared the initial rates of H2O2 generating activity of microsomal preparations containing various human recombinant cytochromes P450s. In the absence of cytochrome P450s the human recombinant NADPH cytochrome P450 reductase (CPR) generated low, but detectable amounts of H2O2 (∼0.04 nmol H2O2/min/100 units of reductase). Significantly greater activity was detected in preparations containing individual cytochrome P450s coexpressed with CPR (from 6.0 nmol H2O2/min/nmol P450 to 0.2 nmol/min/nmol P450); CYP1A1 was the most active, followed by CYP2D6, CYP3A4, CYP2E1, CYP4A11, CYP1A2, and CYP2C subfamily enzymes. H2O2 generating activity of the cytochrome P450s was independent of the ratio of CYP/CPR. Thus, similar H2O2 generating activity was noted with the same cytochrome P450s (CYP3A4, CYP2E1, and CYP2C9) expressed at or near the ratio of CYP/CPR in human liver microsomes (5–7), and when CPR was present in excess (CYP/CPR = 0.2–0.3). Because CYP3A4/5/7 represent up to 40% of total cytochrome P450 in the liver, these data indicate that these enzymes are the major source of H2O2 in human liver microsomes. PMID:25061110

  1. Influences of Surface and Ionic Properties on Electricity Generation of an Active Transducer Driven by Water Motion.

    PubMed

    Park, Junwoo; Yang, YoungJun; Kwon, Soon-Hyung; Kim, Youn Sang

    2015-02-19

    In this Letter, we discuss the surface, ionic properties, and scale-up potential of an active transducer that generated electricity from natural water motion. When a liquid contacts a solid surface, an electrical double layer (EDL) is always formed at the solid/liquid interface. By modulating the EDL, the active transducer could generate a peak voltage of ∼3 V and a peak power of ∼5 μW. Interestingly, there were specific salinities of solution droplets that showed maximum performance and different characteristics according to the ions' nature. Analyzing the results macroscopically, we tried to figure out the origins of the active transducing precipitated by ions dynamics. Also, we demonstrated the scale-up potential for practical usage by multiple electrode design.

  2. 20 CFR 667.262 - Are employment generating activities, or similar activities, allowable under WIA title I?

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... about community job training needs, and (iii) Promote the use of first source hiring agreements and... section, employer outreach and job development activities are directly related to training for eligible individuals. (b) These employer outreach and job development activities include: (1) Contacts with...

  3. 20 CFR 667.262 - Are employment generating activities, or similar activities, allowable under WIA title I?

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... informed decisions about community job training needs, and (iii) Promote the use of first source hiring... this section, employer outreach and job development activities are directly related to training for eligible individuals. (b) These employer outreach and job development activities include: (1) Contacts...

  4. 20 CFR 667.262 - Are employment generating activities, or similar activities, allowable under WIA title I?

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... about community job training needs, and (iii) Promote the use of first source hiring agreements and... section, employer outreach and job development activities are directly related to training for eligible individuals. (b) These employer outreach and job development activities include: (1) Contacts with...

  5. 20 CFR 667.262 - Are employment generating activities, or similar activities, allowable under WIA title I?

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... informed decisions about community job training needs, and (iii) Promote the use of first source hiring... this section, employer outreach and job development activities are directly related to training for eligible individuals. (b) These employer outreach and job development activities include: (1) Contacts...

  6. 20 CFR 667.262 - Are employment generating activities, or similar activities, allowable under WIA title I?

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... informed decisions about community job training needs, and (iii) Promote the use of first source hiring... this section, employer outreach and job development activities are directly related to training for eligible individuals. (b) These employer outreach and job development activities include: (1) Contacts...

  7. Heat shock-induced attenuation of hydroxyl radical generation and mitochondrial aconitase activity in cardiac H9c2 cells.

    PubMed

    Ilangovan, Govindasamy; Venkatakrishnan, C D; Bratasz, Anna; Osinbowale, Sola; Cardounel, Arturo J; Zweier, Jay L; Kuppusamy, Periannan

    2006-02-01

    A mild heat shock (hyperthermia) protects cells from apoptotic and necrotic deaths by inducing overexpression of various heat shock proteins (Hsps). These proteins, in combination with the activation of the nitric oxide synthase (NOS) enzyme, play important roles in the protection of the myocardium against a variety of diseases. In the present work we report that the generation of potent reactive oxygen species (ROS), namely *OH in cardiac H9c2 cells, is attenuated by heat shock treatment (2 h at 42 degrees C). Western blot analyses showed that heat shock treatment induced overexpression of Hsp70, Hsp60, and Hsp25. The observed *OH was found to be derived from the superoxide (O(2)(-)*) generated by the mitochondria. Whereas the manganese superoxide dismutase (MnSOD) activity was increased in the heat-shocked cells, the mitochondrial aconitase activity was reduced. The mechanism of O(2)(-)* conversion into *OH in mitochondria is proposed as follows. The O(2)(-)* leaked from the electron transport chain, oxidatively damages the mitochondrial aconitase, releasing a free Fe(2+). The aconitase-released Fe(2+) combines with H(2)O(2) to generate *OH via a Fenton reaction and the oxidized Fe(3+) recombines with the inactivated enzyme after being reduced to Fe(2+) by other cellular reductants, turning it over to be active. However, in heat-shocked cells, because of higher MnSOD activity, the excess H(2)O(2) causes irreversible damage to the mitochondrial aconitase enzyme, thus inhibiting its activity. In conclusion, we propose that attenuation of *OH generation after heat shock treatment might play an important role in reducing the myocardial ischemic injury, observed in heat shock-treated animals.

  8. Pattern-dependent role of NMDA receptors in action potential generation: consequences on extracellular signal-regulated kinase activation.

    PubMed

    Zhao, Meilan; Adams, J Paige; Dudek, Serena M

    2005-07-27

    Synaptic long-term potentiation is maintained through gene transcription, but how the nucleus is recruited remains controversial. Activation of extracellular signal-regulated kinases (ERKs) 1 and 2 with synaptic stimulation has been shown to require NMDA receptors (NMDARs), yet stimulation intensities sufficient to recruit action potentials (APs) also appear to be required. This has led us to ask the question of whether NMDARs are necessary for AP generation as they relate to ERK activation. To test this, we examined the effects of NMDAR blockade on APs induced with synaptic stimulation using whole-cell current-clamp recordings from CA1 pyramidal cells in hippocampal slices. NMDAR antagonists were found to potently inhibit APs generated with 5 and 100 Hz synaptic stimulation. Blockade of APs and ERK activation could be overcome with the addition of the GABAA antagonist bicuculline, indicating that APs are sufficient to activate signals such as ERK in the nucleus and throughout the neuron in the continued presence of NMDAR antagonists. Interestingly, no effects of the NMDAR antagonists were observed when theta-burst stimulation (TBS) was used. This resistance to the antagonists is conferred by temporal summation during the bursts. These results clarify findings from a previous study showing that ERK activation induced with TBS is resistant to 2-amino-5-phosphonovalerate, in contrast to that induced with 5 or 100 Hz stimulation, which is sensitive. By showing that NMDAR blockade inhibits AP generation, we demonstrate that a major role that NMDARs play in cell-wide and nuclear ERK activation is through their contribution to action potential generation.

  9. NOX4 activity is determined by mRNA levels and reveals a unique pattern of ROS generation

    PubMed Central

    Serrander, Lena; Cartier, Laetitia; Bedard, Karen; Banfi, Botond; Lardy, Bernard; Plastre, Olivier; Sienkiewicz, Andrzej; Fórró, Lászlo; Schlegel, Werner; Krause, Karl-Heinz

    2007-01-01

    NOX4 is an enigmatic member of the NOX (NADPH oxidase) family of ROS (reactive oxygen species)-generating NADPH oxidases. NOX4 has a wide tissue distribution, but the physiological function and activation mechanisms are largely unknown, and its pharmacology is poorly understood. We have generated cell lines expressing NOX4 upon tetracycline induction. Tetracycline induced a rapid increase in NOX4 mRNA (1 h) followed closely (2 h) by a release of ROS. Upon tetracycline withdrawal, NOX4 mRNA levels and ROS release decreased rapidly (<24 h). In membrane preparations, NOX4 activity was selective for NADPH over NADH and did not require the addition of cytosol. The pharmacological profile of NOX4 was distinct from other NOX isoforms: DPI (diphenyleneiodonium chloride) and thioridazine inhibited the enzyme efficiently, whereas apocynin and gliotoxin did not (IC50>100 μM). The pattern of NOX4-dependent ROS generation was unique: (i) ROS release upon NOX4 induction was spontaneous without need for a stimulus, and (ii) the type of ROS released from NOX4-expressing cells was H2O2, whereas superoxide (O2−) was almost undetectable. Probes that allow detection of intracellular O2− generation yielded differential results: DHE (dihydroethidium) fluorescence and ACP (1-acetoxy-3-carboxy-2,2,5,5-tetramethylpyrrolidine) ESR measurements did not detect any NOX4 signal, whereas a robust signal was observed with NBT. Thus NOX4 probably generates O2− within an intracellular compartment that is accessible to NBT (Nitro Blue Tetrazolium), but not to DHE or ACP. In conclusion, NOX4 has a distinct pharmacology and pattern of ROS generation. The close correlation between NOX4 mRNA and ROS generation might hint towards a function as an inducible NOX isoform. PMID:17501721

  10. Bioactive Lysophospholipids Generated by Hepatic Lipase Degradation of Lipoproteins Lead to Complement Activation via the Classical Pathway

    PubMed Central

    Ma, Wanchao; Paik, David C.; Barile, Gaetano R.

    2014-01-01

    Purpose. We determined bioactivity of lysophospholipids generated by degradation of the low-density (LDL), very low-density (VLDL), and high-density (HDL) lipoproteins with hepatic lipase (HL), cholesterol esterase (CE), and lipoprotein-associated phospholipase A2 (Lp-PLA2). Methods. The LDL, VLDL, and HDL were treated with HL, CE, and Lp-PLA2 after immobilization on plates, and complement activation studies were performed with diluted human serum. Complement component 3 (C3) fixation, a marker for complement activation, was determined with a monoclonal anti-human C3d antibody. Enzymatic properties of HL and CE were assayed with triglyceride and phosphatidylcholine substrates for triglyceride hydrolase and phospholipase A activities. The ARPE-19 cells were used for viability studies. Results. The HL degradation of human lipoproteins LDL, VLDL, or HDL results in the formation of modified lipoproteins that can activate the complement pathway. Complement activation is dose- and time-dependent upon HL and occurs via the classical pathway. Enzymatic studies suggest that the phospholipase A1 activity of HL generates complement-activating lysophospholipids. C-reactive protein (CRP), known to simultaneously interact with complement C1 and complement factor H (CFH), further enhances HL-induced complement activation. The lysophospholipids, 1-Palmitoyl-sn-glycero-3-phosphocholine and 1-Oleoyl-sn-glycero-3-phosphocholine, can be directly cytotoxic to ARPE-19 cells. Conclusions. The HL degradation of lipoproteins, known to accumulate in the outer retina and in drusen, can lead to the formation of bioactive lysophospholipids that can trigger complement activation and induce RPE cellular dysfunction. Given the known risk associations for age-related macular degeneration (AMD) with HL, CRP, and CFH, this study elucidates a possible damage pathway for age-related macular degeneration (AMD) in genetically predisposed individuals, that HL activity may lead to accumulation of

  11. SHP-1 overexpression increases the radioresistance of NPC cells by enhancing DSB repair, increasing S phase arrest and decreasing cell apoptosis.

    PubMed

    Pan, Xiaofen; Mou, Jingjing; Liu, Sha; Sun, Ziyi; Meng, Rui; Zhou, Zhenwei; Wu, Gang; Peng, Gang

    2015-06-01

    The present study aimed to investigate the influence of SHP-1 on the radioresistance of the nasopharyngeal carcinoma (NPC) cell line CNE-2 and the relevant underlying mechanisms. The human NPC cell line CNE-2 was transfected with a lentivirus that contained the SHP-1 gene or a nonsense sequence (referred to as LP-H1802Lv201 and LP-NegLv201 cells, respectively). Cells were irradiated with different ionizing radiation (IR) doses. Cell survival, DNA double-strand breaks (DSBs), apoptosis, cell cycle distribution, and the expression of related proteins were assessed using colony formation assay, immunofluorescent assays (IFAs), flow cytometry (FCM) and western blot analyses, respectively. Compared with the control (CNE-2 cells) and LP-NegLv201 cells, LP-H1802Lv201 cells were more resistant to IR. IFAs showed that IR caused less histone H2AX phosphorylation (γH2AX) and RAD51 foci in the LP-H1802Lv201 cells. Compared with the control and LP-NegLv201 cells, LP-H1802Lv201 cells showed increased S phase arrest. After IR, the apoptotic rate of the LP-H1802Lv201 cells was lower in contrast to the control and LP-NegLv201 cells. Western blot analyses showed that IR increased the phosphorylation of ataxia telangiectasia mutated (ATM) kinase, checkpoint kinase 2 (CHK2), ataxia telangiectasia and Rad3-related (ATR) protein, checkpoint kinase 1 (CHK1) and p53. In LP-H1802Lv201 cells, the phosphorylation levels of ATM and CHK2 were significantly increased while the p53 phosphorylation level was decreased compared to these levels in the control and LP-NegLv201 cells. Phosphorylation of ATR and CHK1 did not show significant differences in the three cell groups. Overexpression of SHP-1 in the CNE-2 cells led to radioresistance and the radioresistance was related to enhanced DNA DSB repair, increased S phase arrest and decreased cell apoptosis.

  12. Near-Monodisperse Ni-Cu Bimetallic Nanocrystals of Variable Composition: Controlled Synthesis and Catalytic Activity for H2 Generation

    SciTech Connect

    Zhang, Yawen; Huang, Wenyu; Habas, Susan E.; Kuhn, John N.; Grass, Michael E.; Yamada, Yusuke; Yang, Peidong; Somorjai, Gabor A.

    2008-07-22

    Near-monodisperse Ni{sub 1-x}Cu{sub x} (x = 0.2-0.8) bimetallic nanocrystals were synthesized by a one-pot thermolysis approach in oleylamine/1-octadecene, using metal acetylacetonates as precursors. The nanocrystals form large-area 2D superlattices, and display a catalytic synergistic effect in the hydrolysis of NaBH{sub 4} to generate H{sub 2} at x = 0.5 in a strongly basic medium. The Ni{sub 0.5}Cu{sub 0.5} nanocrystals show the lowest activation energy, and also exhibit the highest H{sub 2} generation rate at 298 K.

  13. The activL® Artificial Disc: a next-generation motion-preserving implant for chronic lumbar discogenic pain

    PubMed Central

    Yue, James J; Garcia, Rolando; Miller, Larry E

    2016-01-01

    Degeneration of the lumbar intervertebral discs is a leading cause of chronic low back pain in adults. Treatment options for patients with chronic lumbar discogenic pain unresponsive to conservative management include total disc replacement (TDR) or lumbar fusion. Until recently, only two lumbar TDRs had been approved by the US Food and Drug Administration − the Charité Artificial Disc in 2004 and the ProDisc-L Total Disc Replacement in 2006. In June 2015, a next-generation lumbar TDR received Food and Drug Administration approval − the activL® Artificial Disc (Aesculap Implant Systems). Compared to previous-generation lumbar TDRs, the activL® Artificial Disc incorporates specific design enhancements that result in a more precise anatomical match and allow a range of motion that better mimics the healthy spine. The results of mechanical and clinical studies demonstrate that the activL® Artificial Disc results in improved mechanical and clinical outcomes versus earlier-generation artificial discs and compares favorably to lumbar fusion. The purpose of this report is to describe the activL® Artificial Disc including implant characteristics, intended use, surgical technique, postoperative care, mechanical testing, and clinical experience to date. PMID:27274317

  14. The activL(®) Artificial Disc: a next-generation motion-preserving implant for chronic lumbar discogenic pain.

    PubMed

    Yue, James J; Garcia, Rolando; Miller, Larry E

    2016-01-01

    Degeneration of the lumbar intervertebral discs is a leading cause of chronic low back pain in adults. Treatment options for patients with chronic lumbar discogenic pain unresponsive to conservative management include total disc replacement (TDR) or lumbar fusion. Until recently, only two lumbar TDRs had been approved by the US Food and Drug Administration - the Charité Artificial Disc in 2004 and the ProDisc-L Total Disc Replacement in 2006. In June 2015, a next-generation lumbar TDR received Food and Drug Administration approval - the activL(®) Artificial Disc (Aesculap Implant Systems). Compared to previous-generation lumbar TDRs, the activL(®) Artificial Disc incorporates specific design enhancements that result in a more precise anatomical match and allow a range of motion that better mimics the healthy spine. The results of mechanical and clinical studies demonstrate that the activL(®) Artificial Disc results in improved mechanical and clinical outcomes versus earlier-generation artificial discs and compares favorably to lumbar fusion. The purpose of this report is to describe the activL(®) Artificial Disc including implant characteristics, intended use, surgical technique, postoperative care, mechanical testing, and clinical experience to date.

  15. Exome and genome sequencing of nasopharynx cancer identifies NF-κB pathway activating mutations

    PubMed Central

    Li, Yvonne Y; Chung, Grace T. Y.; Lui, Vivian W. Y.; To, Ka-Fai; Ma, Brigette B. Y.; Chow, Chit; Woo, John K, S.; Yip, Kevin Y.; Seo, Jeongsun; Hui, Edwin P.; Mak, Michael K. F.; Rusan, Maria; Chau, Nicole G.; Or, Yvonne Y. Y.; Law, Marcus H. N.; Law, Peggy P. Y.; Liu, Zoey W. Y.; Ngan, Hoi-Lam; Hau, Pok-Man; Verhoeft, Krista R.; Poon, Peony H. Y.; Yoo, Seong-Keun; Shin, Jong-Yeon; Lee, Sau-Dan; Lun, Samantha W. M.; Jia, Lin; Chan, Anthony W. H.; Chan, Jason Y. K.; Lai, Paul B. S.; Fung, Choi-Yi; Hung, Suet-Ting; Wang, Lin; Chang, Ann Margaret V.; Chiosea, Simion I.; Hedberg, Matthew L.; Tsao, Sai-Wah; van Hasselt, Andrew C.; Chan, Anthony T. C.; Grandis, Jennifer R.; Hammerman, Peter S.; Lo, Kwok-Wai

    2017-01-01

    Nasopharyngeal carcinoma (NPC) is an aggressive head and neck cancer characterized by Epstein-Barr virus (EBV) infection and dense lymphocyte infiltration. The scarcity of NPC genomic data hinders the understanding of NPC biology, disease progression and rational therapy design. Here we performed whole-exome sequencing (WES) on 111 micro-dissected EBV-positive NPCs, with 15 cases subjected to further whole-genome sequencing (WGS), to determine its mutational landscape. We identified enrichment for genomic aberrations of multiple negative regulators of the NF-κB pathway, including CYLD, TRAF3, NFKBIA and NLRC5, in a total of 41% of cases. Functional analysis confirmed inactivating CYLD mutations as drivers for NPC cell growth. The EBV oncoprotein latent membrane protein 1 (LMP1) functions to constitutively activate NF-κB signalling, and we observed mutual exclusivity among tumours with somatic NF-κB pathway aberrations and LMP1-overexpression, suggesting that NF-κB activation is selected for by both somatic and viral events during NPC pathogenesis. PMID:28098136

  16. Negative correlation between thyroperoxidase and dual oxidase H2O2-generating activities in thyroid nodular lesions.

    PubMed

    Ginabreda, Maria Gloria P; Cardoso, Luciene C; Nobrega, Flavia M; Ferreira, Andrea C F; Gonçalves, Manuel Domingos C; Vaisman, Mário; Carvalho, Denise P

    2008-02-01

    Iodine incorporation into thyroglobulin is dependent on the activities of both thyroperoxidase (TPO) and thyroid dual oxidase 2 (DuOx2). Although TPO expression is decreased in some thyroid nodular lesions, DuOx1 and 2 mRNA expressions are maintained, but DuOx H2O2-generating activity has never been evaluated in such tumors. Our goal was to determine DuOx activity in hypofunctioning lesions of the thyroid. We evaluated H2O2 generation by DuOx in 12 paranodular to cold nodule samples, 17 non-toxic multinodular goiters (MNG; 33 samples), 3 papillary carcinomas (PC; 4 samples), 3 follicular carcinomas (FC; 4 samples), and 10 follicular adenomas. DuOx activity was detected in all paranodular tissues (121+/-23 nmol H2O2/h per mg protein), but was undetectable (<1 nmol H2O2 generated) in all PC, two out of four FC samples and seven out of ten adenomas. In 11 MNG at least two different areas of the goiter have been evaluated, and in 5 of these goiters one of the samples had DuOx activity below the limit of detection. The coefficient of variation in MNG samples ranged from 11.3 to 57.2%. Interestingly, in all the adenomas studied, TPO activity (486+/-142 U/g protein, n=8) was well within the range found in paranodular tissues (414+/-116 U/g protein, n=3). We found a significant negative correlation between DuOx and TPO activities, suggesting that these enzymes are regulated in opposite directions, at least in thyroid tumors.

  17. [Radioisotope thermoelectric generators and ancillary activities]. Monthly technical progress report, 1 April--28 April 1996

    SciTech Connect

    1996-06-01

    Tehnical progress achieved during this period on radioisotope thermoelectric generators is described under the following tasks: engineering support, safety analysis, qualified unicouple fabrication, ETG fabrication/assembly/test, RTG shipping/launch support, design/review/mission applications, and project management/quality assurance/reliability.

  18. In vitro bactericidal activity of aminoglycosides, including the next-generation drug plazomicin, against Brucella spp.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Plazomicin is a next-generation aminoglycoside with a potentially improved safety profile compared to other aminoglycosides. This study assessed plazomicin MICs and MBCs in four Brucella spp. reference strains. Like other aminoglycosides and aminocyclitols, plazomicin MBC values equaled MIC values ...

  19. Influence of Steroids on Oxidant Generation in Activated Human Granulocytes and Mononuclear Leukocytes

    DTIC Science & Technology

    2003-07-01

    and hemorrhage- induced lung injury (31), and lung injury induced by hind limb ischemia (11). Administering scavengers of reactive oxygen species or...be altering protein synthesis. For PMNs, E2 or P4 had no effect on oxidants, whereas all hydro- cortisone concentrations showed a modest trend for...activated and non-activated MNCs (35). Similarly, activated macrophages derived from lungs or a macrophage cell line (J774) treated with pharmacological

  20. Cloud condensation nucleus activity comparison of dry- and wet-generated mineral dust aerosol: the significance of soluble material

    NASA Astrophysics Data System (ADS)

    Garimella, S.; Huang, Y.-W.; Seewald, J. S.; Cziczo, D. J.

    2014-06-01

    This study examines the interaction of clay mineral particles and water vapor for determining the conditions required for cloud droplet formation. Droplet formation conditions are investigated for two common clay minerals, illite and sodium-rich montmorillonite, and an industrially derived sample, Arizona Test Dust. Using wet and dry particle generation coupled to a differential mobility analyzer (DMA) and cloud condensation nuclei counter, the critical activation of the clay mineral particles as cloud condensation nuclei is characterized. Electron microscopy (EM) is used in order to determine non-sphericity in particle shape. It is also used in order to determine particle surface area and account for transmission of multiply charged particles by the DMA. Single particle mass spectrometry and ion chromatography are used to investigate soluble material in wet-generated samples and demonstrate that wet and dry generation yield compositionally different particles. Activation results are analyzed in the context of both κ-Köhler theory (κ-KT) and Frenkel-Halsey-Hill (FHH) adsorption activation theory. This study has two main results: (1) κ-KT is the suitable framework to describe clay mineral nucleation activity. Apparent differences in κ with respect to size arise from an artifact introduced by improper size-selection methodology. For dust particles with mobility sizes larger than ~300 nm, i.e., ones that are within an atmospherically relevant size range, both κ-KT and FHH theory yield similar critical supersaturations. However, the former requires a single hygroscopicity parameter instead of the two adjustable parameters required by the latter. For dry-generated particles, the size dependence of κ is likely an artifact of the shape of the size distribution: there is a sharp drop-off in particle concentration at ~300 nm, and a large fraction of particles classified with a mobility diameter less than ~300 nm are actually multiply charged, resulting in a much

  1. Development of a new deuterium-deuterium (D-D) neutron generator for prompt gamma-ray neutron activation analysis.

    PubMed

    Bergaoui, K; Reguigui, N; Gary, C K; Brown, C; Cremer, J T; Vainionpaa, J H; Piestrup, M A

    2014-12-01

    A new deuterium-deuterium (D-D) neutron generator has been developed by Adelphi Technology for prompt gamma neutron activation analysis (PGNAA), neutron activation analysis (NAA), and fast neutron radiography. The generator makes an excellent fast, intermediate, and thermal neutron source for laboratories and industrial applications that require the safe production of neutrons, a small footprint, low cost, and small regulatory burden. The generator has three major components: a Radio Frequency Induction Ion Source, a Secondary Electron Shroud, and a Diode Accelerator Structure and Target. Monoenergetic neutrons (2.5MeV) are produced with a yield of 10(10)n/s using 25-50mA of deuterium ion beam current and 125kV of acceleration voltage. The present study characterizes the performance of the neutron generator with respect to neutron yield, neutron production efficiency, and the ionic current as a function of the acceleration voltage at various RF powers. In addition the Monte Carlo N-Particle Transport (MCNP) simulation code was used to optimize the setup with respect to thermal flux and radiation protection.

  2. Generation of GFP Reporter Human Induced Pluripotent Stem Cells Using AAVS1 Safe Harbor Transcription Activator-Like Effector Nuclease.

    PubMed

    Luo, Yongquan; Rao, Mahendra; Zou, Jizhong

    2014-05-16

    Generation of a fluorescent GFP reporter line in human induced pluripotent stem cells (hiPSCs) provides enormous potentials in both basic stem cell research and regenerative medicine. A protocol for efficiently generating such an engineered reporter line by gene targeting is highly desired. Transcription activator-like effector nucleases (TALENs) are a new class of artificial restriction enzymes that have been shown to significantly promote homologous recombination by >1000-fold. The AAVS1 (adeno-associated virus integration site 1) locus is a "safe harbor" and has an open chromatin structure that allows insertion and stable expression of transgene. Here, we describe a step-by-step protocol from determination of TALENs activity, hiPSC culture, and delivery of a donor into AAVS1 targeting site, to validation of targeted integration by PCR and Southern blot analysis using hiPSC line, and a pair of open-source AAVS1 TALENs.

  3. Amide-controlled, one-pot synthesis of tri-substituted purines generates structural diversity and analogues with trypanocidal activity.

    PubMed

    Pineda de las Infantas y Villatoro, Maria J; Unciti-Broceta, Juan D; Contreras-Montoya, Rafael; Garcia-Salcedo, Jose A; Gallo Mezo, Miguel A; Unciti-Broceta, Asier; Diaz-Mochon, Juan J

    2015-03-16

    A novel one-pot synthesis of tri-substituted purines and the discovery of purine analogues with trypanocidal activity are reported. The reaction is initiated by a metal-free oxidative coupling of primary alkoxides and diaminopyrimidines with Schiff base formation and subsequent annulation in the presence of large N,N-dimethylamides (e.g. N,N-dimethylpropanamide or larger). This synthetic route is in competition with a reaction previously-reported by our group, allowing the generation of a combinatorial library of tri-substituted purines by the simple modification of the amide and the alkoxide employed. Among the variety of structures generated, two purine analogues displayed trypanocidal activity against the protozoan parasite Trypanosoma brucei with IC50 < 5 μM, being each of those compounds obtained through each of the synthetic pathways.

  4. Project of the borehole neutron generator for the direct determination of oxygen and carbon by activation method

    NASA Astrophysics Data System (ADS)

    Bogdanovich, B. Yu; Vovchenko, E. D.; Iliinskiy, A. V.; Isaev, A. A.; Kozlovskiy, K. I.; Nesterovich, A. V.; Senyukov, V. A.; Shikanov, A. E.

    2016-09-01

    The paper deals with application features of borehole neutron generator (BNG) based on the vacuum accelerating tube (AT) with laser-plasma ion source for determination of oxygen isotope 16O and carbon isotope 12C by direct activation. The project of pulsed BNG for realization of an activation method in the conditions of natural presence of productive hydrocarbons is offered. The diode system with radial acceleration, magnetic electron insulation and laser-plasma source of deuterons at the anode in a sealed-off vacuum accelerating tube is applied. The permanent NdFeB magnet with induction about 0.5 T for produce the insulating magnetic field in the diode gap is proposed. In the experiments on the model of BNG with the accelerating voltage source (≈350 kV), performed by the scheme of Arkadiev-Marx generator, the output of (d, d) neutrons was ∼107 pulse-1.

  5. Comparing Models for Generating a System of Activation and Inhibition of Self-Regulated Learning

    ERIC Educational Resources Information Center

    Magno, Carlo

    2008-01-01

    The study investigated the effect of activation and negative affect on self-regulation. The activation factors are self-determination, disengagement, initiative, and persistence while negative affect is composed of worry, anxiety, thought suppression, and fear of negative evaluation. Separate measures were used for each factor and administered to…

  6. The Need for a Harmonized Repository for Next-Generation Human Activity Data

    EPA Science Inventory

    Multi-tiered human time-activity-location data can inform many efforts to describe human exposures to air pollutants and other chemicals on a range of temporal and spatial scales. In the last decade, EPA's Consolidated Human Activity Database (CHAD) has served as a harmonized rep...

  7. A New Generation of Leaders for Eastern Europe: Values and Attitudes for Active Citizenship

    ERIC Educational Resources Information Center

    Humphreys, Melanie J.

    2011-01-01

    Higher education has a crucial role in developing active citizenship within students. Even though active citizenship skills and competencies have been identified as essential life skills for Europe there is very little progress to date in establishing measures or indicators as to how this might be accomplished. This study provides the first data…

  8. Generating Effective Facilitation Questions for Team-Building/Personal-Challenge Activities

    ERIC Educational Resources Information Center

    Gabbei, Ritchie

    2004-01-01

    Team-building/personal-challenge (TB/PC) activities have become popular ways to address students' interpersonal and intrapersonal skills and abilities associated with the affective domain. The outcomes associated with TB/PC activities are often best experienced and learned through the use of indirect methods of instruction. Typically, many…

  9. New neurons generated from running are broadly recruited into neuronal activation associated with three different hippocampus-involved tasks.

    PubMed

    Clark, Peter J; Bhattacharya, Tushar K; Miller, Daniel S; Kohman, Rachel A; DeYoung, Erin K; Rhodes, Justin S

    2012-09-01

    Running increases the formation of new neurons in the adult rodent hippocampus. However, the function of new neurons generated from running is currently unknown. One hypothesis is that new neurons from running contribute to enhanced cognitive function by increasing plasticity in the adult hippocampus. An alternative hypothesis is that new neurons generated from running incorporate into experience-specific hippocampal networks that only become active during running. The purpose of this experiment was to determine if new neurons generated from running are selectively activated by running, or can become recruited into granule cell activity occurring during performance on other behavioral tasks that engage the hippocampus. Therefore, the activation of new 5-6 week neurons was detected using BrdU, NeuN, and Zif268 triple-label immunohistochemistry in cohorts of female running and sedentary adult C57BL/6J mice following participation in one of three different tasks: the Morris water maze, novel environment exploration, or wheel running. Results showed that running and sedentary mice displayed a nearly equivalent proportion of new neurons that expressed Zif268 following each task. Since running approximately doubled the number of new neurons, the results demonstrated that running mice had a greater number of new neurons recruited into the Zif268 induction in the granule cell layer following each task than sedentary mice. The results suggest that new neurons incorporated into hippocampal circuitry from running are not just activated by wheel running itself, but rather become broadly recruited into granule cell layer activity during distinct behavioral experiences.

  10. Inorganic nitrite attenuates NADPH oxidase-derived superoxide generation in activated macrophages via a nitric oxide-dependent mechanism.

    PubMed

    Yang, Ting; Peleli, Maria; Zollbrecht, Christa; Giulietti, Alessia; Terrando, Niccolo; Lundberg, Jon O; Weitzberg, Eddie; Carlström, Mattias

    2015-06-01

    Oxidative stress contributes to the pathogenesis of many disorders, including diabetes and cardiovascular disease. Immune cells are major sources of superoxide (O2(∙-)) as part of the innate host defense system, but exaggerated and sustained O2(∙-) generation may lead to progressive inflammation and organ injuries. Previous studies have proven organ-protective effects of inorganic nitrite, a precursor of nitric oxide (NO), in conditions manifested by oxidative stress and inflammation. However, the mechanisms are still not clear. This study aimed at investigating the potential role of nitrite in modulating NADPH oxidase (NOX) activity in immune cells. Mice peritoneal macrophages or human monocytes were activated by lipopolysaccharide (LPS), with or without coincubation with nitrite. O2(∙-) and peroxynitrite (ONOO(-)) formation were detected by lucigenin-based chemiluminescence and fluorescence techniques, respectively. The intracellular NO production was measured by DAF-FM DA fluorescence. NOX isoforms and inducible NO synthase (iNOS) expression were detected by qPCR. LPS increased both O2(∙-) and ONOO(-) production in macrophages, which was significantly reduced by nitrite (10µmol/L). Mechanistically, the effects of nitrite are (1) linked to increased NO generation, (2) similar to that observed with the NO donor DETA-NONOate, and (3) can be abolished by the NO scavenger carboxy-PTIO or by the xanthine oxidase (XO) inhibitor febuxostat. Nox2 expression was increased in activated macrophages, but was not influenced by nitrite. However, nitrite attenuated LPS-induced upregulation of iNOS expression. Similar to that observed in mice macrophages, nitrite also reduced O2(∙-) generation in LPS-activated human monocytes. In conclusion, XO-mediated reduction of nitrite attenuates NOX activity in activated macrophages, which may modulate the inflammatory response.

  11. Manassantin A isolated from Saururus chinensis inhibits 5-lipoxygenase-dependent leukotriene C4 generation by blocking mitogen-activated protein kinase activation in mast cells.

    PubMed

    Kim, Su Jeong; Lu, Yue; Kwon, Okyun; Hwangbo, Kyoung; Seo, Chang-Seob; Lee, Seung Ho; Kim, Cheorl-Ho; Chang, Young-Chae; Son, Jong Keun; Chang, Hyeun Wook

    2011-01-01

    In this study, manassantin A (Man A), an herbal medicine isolated from Saururus chinensis (S. chinensis), markedly inhibited 5-lipoxygenase (5-LO)-dependent leukotriene C(4) (LTC(4)) generation in bone marrow-derived mast cells (BMMCs) in a concentration-dependent manner. To investigate the molecular mechanisms underlying the inhibition of LTC(4) generation by Man A, we assessed the effects of Man A on phosphorylation of cytosolic phospholipase A(2) (cPLA(2)) and mitogen-activated protein kinases (MAPKs). Inhibition of LTC(4) generation by Man A was accompanied by a decrease in cPLA(2) phosphorylation, which occurred via the MAPKs including extracellular signal-regulated protein kinase-1/2 (ERK1/2) as well as p38 and c-Jun N-terminal kinase (JNK) pathways. Taken together, the present study suggests the Man A represents a potential therapeutic approach for the treatment of airway allergic-inflammatory diseases.

  12. Endogenous platelet factor 4 stimulates activated protein C generation in vivo and improves survival after thrombin or lipopolysaccharide challenge

    PubMed Central

    Mahmud, Shawn A.; Lambert, Michele P.; Poncz, Mortimer; Slungaard, Arne

    2007-01-01

    Pharmacologic infusion of activated protein C (APC) improves survival in severe sepsis, and platelet factor 4 (PF4) accelerates APC generation in a primate thrombin-infusion model. We now tested whether endogenous platelet PF4 content affects APC generation. Mice completely deficient in PF4 (mPF4−/−) had impaired APC generation and survival after thrombin infusion, similar to the impairment seen in heterozygote protein C–deficient (PC+/−) mice. Transgenic mice overexpressing human PF4 (hPF4+) had increased plasma APC generation. Overexpression of platelet PF4 compensated for the defect seen in PC+/− mice. In both a thrombin and a lipopolysaccharide (LPS) survival model, hPF4+ and PC+/−/hPF4+ mice had improved survival. Further, infusion of hPF4+ platelets improved survival of wild-type mice after an LPS challenge. These studies suggest that endogenous PF4 release may have biologic consequences for APC generation and survival in clinical sepsis. Infusions of PF4-rich platelets may be an effective strategy to improve outcome in this setting. PMID:17540840

  13. Endogenous platelet factor 4 stimulates activated protein C generation in vivo and improves survival after thrombin or lipopolysaccharide challenge.

    PubMed

    Kowalska, M Anna; Mahmud, Shawn A; Lambert, Michele P; Poncz, Mortimer; Slungaard, Arne

    2007-09-15

    Pharmacologic infusion of activated protein C (APC) improves survival in severe sepsis, and platelet factor 4 (PF4) accelerates APC generation in a primate thrombin-infusion model. We now tested whether endogenous platelet PF4 content affects APC generation. Mice completely deficient in PF4 (mPF4(-/-)) had impaired APC generation and survival after thrombin infusion, similar to the impairment seen in heterozygote protein C-deficient (PC(+/-)) mice. Transgenic mice overexpressing human PF4 (hPF4(+)) had increased plasma APC generation. Overexpression of platelet PF4 compensated for the defect seen in PC(+/-) mice. In both a thrombin and a lipopolysaccharide (LPS) survival model, hPF4(+) and PC(+/-)/hPF4(+) mice had improved survival. Further, infusion of hPF4(+) platelets improved survival of wild-type mice after an LPS challenge. These studies suggest that endogenous PF4 release may have biologic consequences for APC generation and survival in clinical sepsis. Infusions of PF4-rich platelets may be an effective strategy to improve outcome in this setting.

  14. Performance Analysis of Positive-feedback-based Active Anti-islanding Schemes for Inverter-Based Distributed Generators

    SciTech Connect

    Du, Pengwei; Aponte, Erick E.; Nelson, J. Keith

    2010-06-14

    Recently proposed positive-feedback-based anti-islanding schemes (AI) are highly effective in preventing islanding without causing any degradation in power quality. This paper aims to analyze the performance of these schemes quantitatively in the context of the dynamic models of inverter-based distributed generators (DG). In this study, the characteristics of these active anti-islanding methods are discussed and design guidelines are derived.

  15. Soliton generation from an actively mode-locked fiber laser incorporating an electro-optic fiber modulator.

    PubMed

    Malmström, Mikael; Margulis, Walter; Tarasenko, Oleksandr; Pasiskevicius, Valdas; Laurell, Fredrik

    2012-01-30

    This work demonstrates an actively mode-locked fiber laser operating in soliton regime and employing an all-fiber electro-optic modulator. Nonlinear polarization rotation is utilized for femtosecond pulse generation. Stable operation of the all-fiber ring laser is readily achieved at a fundamental repetition rate of 2.6 MHz and produces 460 fs pulses with a spectral bandwidth of 5.3 nm.

  16. Generation of spectral clusters in a mixture of noble and Raman-active gases

    NASA Astrophysics Data System (ADS)

    Hosseini, Pooria; Abdolvand, Amir; St. J. Russell, Philip

    2016-12-01

    We report a novel scheme for the generation of dense clusters of Raman sidebands. The scheme uses a broadband-guiding hollow-core photonic crystal fiber (HC-PCF) filled with a mixture of H2, D2, and Xe for efficient interaction between the gas mixture and a green laser pump pulse (532 nm, 1 ns) of only 5 uJ energy. This results in the generation from noise of more than 135 ro-vibrational Raman sidebands covering the visible spectral region with an average spacing of only 2 THz. Such a spectrally dense and compact fiber-based source is ideal for applications where closely spaced narrow-band laser lines with high spectral power density are required, such as in spectroscopy and sensing. When the HC-PCF is filled with a H2-D2 mixture the Raman comb spans the spectral region from the deep UV (280 nm) to the near infrared (1000 nm).

  17. Skin tumor-promoting activity of benzoyl peroxide, a widely used free radical-generating compound

    SciTech Connect

    Slaga, T.J.; Klein-Szanto, A.J.P.; Triplett, L.L.; Yotti, L.P.; Trosko, J.E.

    1981-08-28

    Benzoyl peroxide, a widely used free radical-generating compound, promoted both papillomas and carcinomas when it was topically applied to mice after 7, 12-dimethylbenz(a)anthracene initiation. Benzoyl peroxide was inactive on the skin as a complete carcinogen or as a tumor initiator. A single topical application of benzoyl peroxide produced a marked epidermal hyperplasia and induced a large number of dark basal keratinocytes, effects similar to those produced by the potent tumor promoter 12-O-tetradecanoyl phorbol-13-acetate. Benzoyl peroxide, like other known tumor promoters, also inhibited metabolic cooperation (intercellular communication) in Chinese hamster cells. In view of these results caution should be recommended in the use of this and other free radical-generating compounds.

  18. Building the Next Generation of Scientific Explorers through Active Engagement with STEM Experts and International Space Station Resources

    NASA Technical Reports Server (NTRS)

    Graff, P. V.; Vanderbloemen, L.; Higgins, M.; Stefanov, W. L.; Rampe, E.

    2015-01-01

    Connecting students and teachers in classrooms with science, technology, engineering, and mathematics (STEM) experts provides an invaluable opportunity for all. These experts can share the benefits and utilization of resources from the International Space Station (ISS) while sharing and "translating" exciting science being conducted by professional scientists. Active engagement with these STEM experts involves students in the journey of science and exploration in an enthralling and understandable manner. This active engagement, connecting classrooms with scientific experts, helps inspire and build the next generation of scientific explorers in academia, private industry, and government.

  19. Active Mirrors for On-Orbit Alignment of the Second Generation Wide Field and Planetary Camera

    NASA Technical Reports Server (NTRS)

    Fanson, J.; Trauger, J.

    1993-01-01

    This paper describes the assembly level testing and calibration of the Articulating Fold Mirrors (AFMs), and the development of a software tool that generates the commands for adjusting the positions of the Pickoff Mirror Mechanism (PMM) and the AFMs to achieve system level optical alignment. Our experience with the POMM and AFMs through system level calibration and testing of the WFPC-2 instrument is described.

  20. Central station applications planning activities and supporting studies. [application of photovoltaic technology to power generation plants

    NASA Technical Reports Server (NTRS)

    Leonard, S. L.; Siegel, B.

    1980-01-01

    The application of photovoltaic technology in central station (utility) power generation plants is considered. A program of data collection and analysis designed to provide additional information about the subset of the utility market that was identified as the initial target for photovoltaic penetration, the oil-dependent utilities (especially muncipals) of the U.S. Sunbelt, is described along with a series of interviews designed to ascertain utility industry opinions about the National Photovoltaic Program as it relates to central station applications.

  1. Pore development in carbonized hemoglobin by concurrently generated MgO template for activity enhancement as fuel cell cathode catalyst.

    PubMed

    Maruyama, Jun; Hasegawa, Takahiro; Amano, Taiji; Muramatsu, Yasuji; Gullikson, Eric M; Orikasa, Yuki; Uchimoto, Yoshiharu

    2011-12-01

    Various carbon materials with a characteristic morphology and pore structure have been produced using template methods in which a carbon-template composite is once formed and the characteristic features derived from the template are generated after the template removal. In this study, hemoglobin, which is a natural compound that could be abundantly and inexpensively obtained, was used as the carbon material source to produce a carbonaceous noble-metal-free fuel cell cathode catalyst. Magnesium oxide was used as the template concurrently generated with the hemoglobin carbonization from magnesium acetate mixed with hemoglobin as the starting material mixture to enable pore development for improving the activity of the carbonized hemoglobin for the cathodic oxygen reduction. After removal of the MgO template, the substantially developed pores were generated in the carbonized hemoglobin with an amorphous structure observed by total-electron-yield X-ray absorption. The extended X-ray absorption fine structure at the Fe-K edge indicated that Fe was coordinated with four nitrogen atoms (Fe-N(4) moiety) in the carbonized hemoglobin. The oxygen reduction activity of the carbonized hemoglobin evaluated using rotating disk electrodes was dependent on the pore structure. The highly developed pores led to an improved activity.

  2. Substrate shuttling between active sites of uroporphyrinogen decarboxylase is not required to generate coproporphyrinogen

    PubMed Central

    Phillips, John D.; Warby, Christy A.; Whitby, Frank G.; Kushner, James P.; Hill, Christopher P.

    2009-01-01

    Summary Uroporphyrinogen Decarboxylase (URO-D; EC 4.1.1.37), the fifth enzyme of the heme biosynthetic pathway, is required for the production of heme, vitamin B12, siroheme, and chlorophyll precursors. URO-D catalyzes the sequential decarboxylation of the four acetate side chains on the pyrrole groups of uroporphyrinogen to produce coproporphyrinogen. URO-D is a stable homodimer with the active site clefts of the two subunits adjacent to each other. It has been hypothesized that the two catalytic centers interact functionally, perhaps by shuttling of reaction intermediates between subunits. We tested this hypothesis by construction of a single chain protein (scURO-D) in which the two subunits were connected by a flexible linker. The crystal structure of this protein was shown to be superimposible with wild-type activity and have comparable catalytic activity. Mutations that impaired one or the other of the two active sites of scURO-D resulted in approximately half of wild-type activity. The distribution of reaction intermediates was the same for mutant and wild-type sequences, and was unaltered in a competition experiment using the I and III isomer substrates. These observations indicate that communication between active sites is not required for enzyme function, and suggest that the dimeric structure of URO-D is required to achieve conformational stability and create a large active site cleft. PMID:19362562

  3. Substrate Shuttling Between Active Sites of Uroporphyrinogen Decarboxylase in Not Required to Generate Coproporphyrinogen

    SciTech Connect

    Phillips, J.; Warby, C; Whitby, F; Kushner, J; Hill, C

    2009-01-01

    Uroporphyrinogen decarboxylase (URO-D; EC 4.1.1.37), the fifth enzyme of the heme biosynthetic pathway, is required for the production of heme, vitamin B12, siroheme, and chlorophyll precursors. URO-D catalyzes the sequential decarboxylation of four acetate side chains in the pyrrole groups of uroporphyrinogen to produce coproporphyrinogen. URO-D is a stable homodimer, with the active-site clefts of the two subunits adjacent to each other. It has been hypothesized that the two catalytic centers interact functionally, perhaps by shuttling of reaction intermediates between subunits. We tested this hypothesis by construction of a single-chain protein (single-chain URO-D) in which the two subunits were connected by a flexible linker. The crystal structure of this protein was shown to be superimposable with wild-type activity and to have comparable catalytic activity. Mutations that impaired one or the other of the two active sites of single-chain URO-D resulted in approximately half of wild-type activity. The distributions of reaction intermediates were the same for mutant and wild-type sequences and were unaltered in a competition experiment using I and III isomer substrates. These observations indicate that communication between active sites is not required for enzyme function and suggest that the dimeric structure of URO-D is required to achieve conformational stability and to create a large active-site cleft.

  4. Engineering of a linear inactive analog of human β-defensin 4 to generate peptides with potent antimicrobial activity.

    PubMed

    Sharma, Himanshu; Mathew, Basil; Nagaraj, Ramakrishnan

    2015-06-01

    Human β-defensins (HBDs) are cationic antimicrobial peptides constrained by three disulfide bridges. They have diverse range of functions in the innate immune response. It is of interest to investigate whether linear analogs of defensins can be generated, which possess antimicrobial activity. In this study, we have designed linear peptides with potent antimicrobial activity from an inactive peptide spanning the N-terminus of HBD4. Our results show that l-arginine to d-arginine substitution imparts considerable antimicrobial activity against both bacteria and Candida albicans. Increase in hydrophobicity by fatty acylation of the peptides with myristic acid further enhances their potency. In the presence of high concentrations of salt, antimicrobial activity of the myristoylated peptide with l-arginine is attenuated relatively to a lesser extent as compared with the linear active peptide with d-arginine. Substitution of cysteine with the hydrophobic helix-promoting amino acid α-aminoisobutyric acid favors candidacidal activity but not antibacterial activity. The mechanism of killing by d-arginine substituted unacylated analog involves transient interaction with the bacterial membrane followed by translocation into the cytoplasm without membrane permeabilization. Accumulation of peptides in the cytoplasm can affect various cellular processes that lead to cell death. However, the peptide causes membrane permeabilization in case of C. albicans. Myristoylation results in greater interaction of the peptide chain with the microbial cell surface and causes membrane permeabilization. Results described in the study demonstrate that it is possible to generate highly active linear analogs of defensins by selective introduction of d-amino acids and fatty acids, which could be attractive candidates for development as therapeutic agents.

  5. Transparency in a Pediatric Quality Improvement Collaborative: A Passionate Journey by NPC-QIC Clinicians and Parents.

    PubMed

    Lihn, Stacey L; Kugler, John D; Peterson, Laura E; Lannon, Carole M; Pickles, Diane; Beekman, Robert H

    2015-01-01

    Transparency-sharing data or information about outcomes, processes, protocols, and practices-may be the most powerful driver of health care improvement. In this special article, the development and growth of transparency within the National Pediatric Cardiology Quality Improvement Collaborative is described. The National Pediatric Cardiology Quality Improvement Collaborative transparency journey is guided by equal numbers of clinicians and parents of children with congenital heart disease working together in a Transparency Work Group. Activities are organized around four interrelated levels of transparency (individual, organizational, collaborative, and system), each with a specified purpose and aim. A number of Transparency Work Group recommendations have been operationalized. Aggregate collaborative performance is now reported on the public-facing web site. Specific information that the Transparency Work Group recommends centers provide to parents has been developed and published. Almost half of National Pediatric Cardiology Quality Improvement Collaborative centers participated in a pilot of transparently sharing their outcomes achieved with one another. Individual centers have also begun successfully implementing recommended transparency activities. Despite progress, barriers to full transparency persist, including health care organization concerns about potential negative effects of disclosure on reputation and finances, and lack of reliable definitions, data, and reporting standards for fair comparisons of centers. The National Pediatric Cardiology Quality Improvement Collaborative's transparency efforts have been a journey that continues, not a single goal or destination. Balanced participation of clinicians and parents has been a critical element of the collaborative's success on this issue. Plans are in place to guide implementation of additional transparency recommendations across all four levels, including extension of the activities beyond the

  6. Artocarpol A stimulation of superoxide anion generation in neutrophils involved the activation of PLC, PKC and p38 mitogen-activated PK signaling pathways.

    PubMed

    Kuan, Yu-Hsiang; Lin, Ruey-Hseng; Tsao, Lo-Ti; Lin, Chun-Nan; Wang, Jih-Pyang

    2005-06-01

    1 Artocarpol A (ART), a natural phenolic compound isolated from Artocarpus rigida, stimulated a slow onset and long-lasting superoxide anion generation in rat neutrophils, whereas only slightly activated the NADPH oxidase in a cell-free system. 2 Pretreatment of neutrophils with pertussis toxin (1 microg ml(-1)), 50 microM 2'-amino-3'-methoxyflavone (PD 98059), or 1 microM 1,4-diamino-2,3-dicyano-1,4-bis(2-aminophenylthio)butadiene (U0126) had no effect on ART-stimulated superoxide anion generation. ART (30 microM) did not induce extracellular signal-regulated kinase (ERK) phosphorylation. 3 4-(4-Fluorophenyl)-2-(4-methylsulfinylphenyl)-5-(4-pyridyl)-1H-imidazole (SB 203580) markedly attenuated the ART-stimulated superoxide anion generation (IC50 value of 4.3+/-0.3 microM). Moreover, ART induced p38 mitogen-activated PK (MAPK) phosphorylation and activation. 4 The superoxide anion generation in response to ART was also substantially inhibited in a Ca2+-free medium, and by pretreatment with 1 microM 1-[6-((17beta-3-methoxyestra-1,3,5(10)-trien-17-yl)amino)hexyl]-1H-pyrrole-2,5-dione (U-73122) and 100 microM 2-aminoethyldiphenyl borate (2-APB). ART (30 microM) stimulated the [Ca2+]i elevation in the presence or absence of external Ca2+, and also increased the D-myo-inositol 1,4,5-trisphosphate formation. 5 2-[1-(3-Dimethylaminopropyl)-1H-indol-3-yl]-3-(1H-indol-3-yl)-maleimide (GF 109203X) greatly inhibited the ART-stimulated superoxide anion generation (IC50 value of 7.8+/-1.0 nM). ART increased the recruitment of PKC-alpha, -betaI, and -betaII to the plasma membrane of neutrophils, and stimulated Ca2+-dependent PKC activation in the cytosol preparation. 6 ART induced the phosphorylation of p47phox, which was attenuated by GF 109203X. Moreover, ART evoked the membrane association of p47(phox), which was inhibited by GF 109203X and SB 203580. 7 These results indicate that the ART stimulation of superoxide anion generation involved the activation of p38 MAPK, PLC/Ca2

  7. Flow-Through, Low Retention Hydrocarbon Generation in Active Sedimentary Basins

    NASA Astrophysics Data System (ADS)

    Cathles, L. M.

    2002-12-01

    Hydrocarbons are typically generated when thin (<100 m), organic-rich strata are buried. Oil is generated first, then gas. Detailed investigation of the chemistry of hydrocarbons in a 100 (E-W) x 200 (N-S) km study area in the offshore Louisiana Gulf of Mexico Basin shows that the early-generated oils there have been extensively altered by interaction with late-generated gas, producing a remarkably regular pattern in which over 90 wt% of the n-alkanes have been removed by gas washing in the north while none have been so removed in the south. Straight-forward (Turcottian) physical-chemical modeling suggests this pattern can be produced only if the retention of hydrocarbons between source and seafloor is very low (0.1% of the pore space). This low retention means that about 80% or 120 billion tones (Bt) of the ~149 Bt of hydrocarbons that have been generated and expelled from the source strata in the study area have been vented into the ocean. This mass of vented hydrocarbons is 100 times the mass of hydrocarbon in known reservoirs in the area (1.4 Bt or 11 billon barrels of oil and gas equivalent). The reservoired hydrocarbons are about 5% of the hydrocarbons retained in the models between the source and seafloor. To be properly washed, the reservoired hydrocarbons must also have been very recently introduced. The hydrocarbon system in the study area is thus one of massive venting with minor, constantly replenished (flow through) retention. The predicted current rates of venting are confirmed by independent estimates of the rate of hydrate accumulation in the southern part of the study area. The massive venting of over 900 billion barrels (equivalent) of hydrocarbons, an amount about equal to the total world consumption of oil to date, from a 100 x 200 km portion of one basin in a relatively short timespan (about 20 Ma) provides an interesting geologic context for human production and consumption. The rapid venting also has potentially important implications for the

  8. Enhanced microbial coalbed methane generation: A review of research, commercial activity, and remaining challenges

    USGS Publications Warehouse

    Ritter, Daniel J.; Vinson, David S.; Barnhart, Elliott P.; Akob, Denise M.; Fields, Matthew W.; Cunningham, Al B.; Orem, William H.; McIntosh, Jennifer C.

    2015-01-01

    Coalbed methane (CBM) makes up a significant portion of the world’s natural gas resources. The discovery that approximately 20% of natural gas is microbial in origin has led to interest in microbially enhanced CBM (MECoM), which involves stimulating microorganisms to produce additional CBM from existing production wells. This paper reviews current laboratory and field research on understanding processes and reservoir conditions which are essential for microbial CBM generation, the progress of efforts to stimulate microbial methane generation in coal beds, and key remaining knowledge gaps. Research has been primarily focused on identifying microbial communities present in areas of CBM generation and attempting to determine their function, in-situ reservoir conditions that are most favorable for microbial CBM generation, and geochemical indicators of metabolic pathways of methanogenesis (i.e., acetoclastic or hydrogenotrophic methanogenesis). Meanwhile, researchers at universities, government agencies, and companies have focused on four primary MECoM strategies: 1) microbial stimulation (i.e., addition of nutrients to stimulate native microbes); 2) microbial augmentation (i.e., addition of microbes not native to or abundant in the reservoir of interest); 3) physically increasing microbial access to coal and distribution of amendments; and 4) chemically increasing the bioavailability of coal organics. Most companies interested in MECoM have pursued microbial stimulation: Luca Technologies, Inc., successfully completed a pilot scale field test of their stimulation strategy, while two others, Ciris Energy and Next Fuel, Inc., have undertaken smaller scale field tests. Several key knowledge gaps remain that need to be addressed before MECoM strategies can be implemented commercially. Little is known about the bacterial community responsible for coal biodegradation and how these microorganisms may be stimulated to enhance microbial methanogenesis. In addition, research

  9. Mutualistic Benefits Generate an Unequal Distribution of Risky Activities Among Unrelated Group Members

    NASA Astrophysics Data System (ADS)

    Kukuk, Penelope F.; Ward, Seamus A.; Jozwiak, Amy

    Recent studies provide a new challenge to the adequacy of theories concerning the evolution of cooperation among nonrelatives: some individuals perform high-risk activities while others do not. We examined a communal hymenopteran species, Lasioglossum(Chilalictus)hemichalceum, to determine why group members engaged in demonstrably risky activities (foraging) tolerate the selfish behavior (remaining in the nest) of unrelated nestmates. Experimental removal of adult females indicated that their presence is required for the protection of brood from ant predators. Nonforagers ensure the continued presence of adults in the nest if the risk-taking foragers die, thereby safeguarding the survival of forager offspring. This results in an unequal distribution of risky activities within social groups in which avoidance of risky activities by some group members is ultimately beneficial to risk takers.

  10. Activity Recognition Using Hybrid Generative/Discriminative Models on Home Environments Using Binary Sensors

    PubMed Central

    Ordóñez, Fco. Javier; de Toledo, Paula; Sanchis, Araceli

    2013-01-01

    Activities of daily living are good indicators of elderly health status, and activity recognition in smart environments is a well-known problem that has been previously addressed by several studies. In this paper, we describe the use of two powerful machine learning schemes, ANN (Artificial Neural Network) and SVM (Support Vector Machines), within the framework of HMM (Hidden Markov Model) in order to tackle the task of activity recognition in a home setting. The output scores of the discriminative models, after processing, are used as observation probabilities of the hybrid approach. We evaluate our approach by comparing these hybrid models with other classical activity recognition methods using five real datasets. We show how the hybrid models achieve significantly better recognition performance, with significance level p < 0.05, proving that the hybrid approach is better suited for the addressed domain. PMID:23615583

  11. Neural activity underlying tinnitus generation: results from PET and fMRI.

    PubMed

    Lanting, C P; de Kleine, E; van Dijk, P

    2009-09-01

    Tinnitus is the percept of sound that is not related to an acoustic source outside the body. For many forms of tinnitus, mechanisms in the central nervous system are believed to play an important role in the pathology. Specifically, three mechanisms have been proposed to underlie tinnitus: (1) changes in the level of spontaneous neural activity in the central auditory system, (2) changes in the temporal pattern of neural activity, and (3) reorganization of tonotopic maps. The neuroimaging methods fMRI and PET measure signals that presumably reflect the firing rates of multiple neurons and are assumed to be sensitive to changes in the level of neural activity. There are two basic paradigms that have been applied in functional neuroimaging of tinnitus. Firstly, sound-evoked responses as well as steady state neural activity have been measured to compare tinnitus patients to healthy controls. Secondly, paradigms that involve modulation of tinnitus by a controlled stimulus allow for a within-subject comparison that identifies neural activity that may be correlated to the tinnitus percept. Even though there are many differences across studies, the general trend emerging from the neuroimaging studies, is that tinnitus in humans may correspond to enhanced neural activity across several centers of the central auditory system. Also, neural activity in non-auditory areas including the frontal areas, the limbic system and the cerebellum seems associated with the perception of tinnitus. These results indicate that in addition to the auditory system, non-auditory systems may represent a neural correlate of tinnitus. Although the currently published neuroimaging studies typically show a correspondence between tinnitus and enhanced neural activity, it will be important to perform future studies on subject groups that are closely matched for characteristics such as age, gender and hearing loss in order to rule out the contribution of these factors to the abnormalities specifically

  12. The influence of Erythropoietin on platelet activation, thrombin generation and FVII/active FVII in patients with AMI

    PubMed Central

    2014-01-01

    Background Erythropoietin (Epo) has been shown to improve myocardial function in models of experimental myocardial infarction, but has also been associated with a rise in thromboembolic events. Thus, the aim of this study was to investigate the influence of Epo on platelet activation and coagulation in patients with acute myocardial infarction (AMI). Methods The study was designed as a substudy of the randomised, double-blind, placebo controlled REVIVAL-3 (REgeneration of VItal Myocardium in ST-Segment EleVation MyocardiAL Infarction by Erythropoietin) study that investigated the effects of recombinant human Epo in AMI. Serial venous blood samples were collected before and after study medication. Circulating prothrombin fragment F1 + 2, FVII, active FVII, beta thromboglobulin (TG) and P-Selectin were measured before and 60 hours after randomization by immunoassay (n = 94). In a randomly selected subgroup platelet aggregation was measured using whole blood aggregometry (Multiplate Analyzer, n = 45). Results After 5 days an increase in FVII was observed after Epo as compared to placebo (P = 0.02), yet active FVII and prothrombin fragment F1 + 2 remained unchanged. Moreover, no statistically significant differences in circulating TG or P-selectin were observed between the groups. As an expected response to peri-interventional therapy with clopidogrel and aspirin, platelet aggregation after stimulation with ADP, TRAP, ASPI or collagen decreased 12 hours and 2 days after PCI. However, no difference between the Epo and the placebo group was observed. Conclusion After treatment with Epo in patients with AMI a slight increase in circulating FVII after Epo was not associated with an increase in active FVII, prothrombin fragment F1 + 2, TG or P-selectin. Moreover, platelet aggregation was not altered after treatment with Epo as compared to placebo. Trial registration ClinicalTrials.gov Identifier: NCT01761435 PMID:25228850

  13. Correlations between prefrontal neurons form a small-world network that optimizes the generation of multineuron sequences of activity.

    PubMed

    Luongo, Francisco J; Zimmerman, Chris A; Horn, Meryl E; Sohal, Vikaas S

    2016-05-01

    Sequential patterns of prefrontal activity are believed to mediate important behaviors, e.g., working memory, but it remains unclear exactly how they are generated. In accordance with previous studies of cortical circuits, we found that prefrontal microcircuits in young adult mice spontaneously generate many more stereotyped sequences of activity than expected by chance. However, the key question of whether these sequences depend on a specific functional organization within the cortical microcircuit, or emerge simply as a by-product of random interactions between neurons, remains unanswered. We observed that correlations between prefrontal neurons do follow a specific functional organization-they have a small-world topology. However, until now it has not been possible to directly link small-world topologies to specific circuit functions, e.g., sequence generation. Therefore, we developed a novel analysis to address this issue. Specifically, we constructed surrogate data sets that have identical levels of network activity at every point in time but nevertheless represent various network topologies. We call this method shuffling activity to rearrange correlations (SHARC). We found that only surrogate data sets based on the actual small-world functional organization of prefrontal microcircuits were able to reproduce the levels of sequences observed in actual data. As expected, small-world data sets contained many more sequences than surrogate data sets with randomly arranged correlations. Surprisingly, small-world data sets also outperformed data sets in which correlations were maximally clustered. Thus the small-world functional organization of cortical microcircuits, which effectively balances the random and maximally clustered regimes, is optimal for producing stereotyped sequential patterns of activity.

  14. Sinomenine induces the generation of intestinal Treg cells and attenuates arthritis via activation of aryl hydrocarbon receptor.

    PubMed

    Tong, Bei; Yuan, Xusheng; Dou, Yannong; Wu, Xin; Wang, Yuhui; Xia, Yufeng; Dai, Yue

    2016-10-01

    Sinomenine (SIN), an anti-arthritis drug, has previously been proven to exert immunomodulatory activity in rats by inducing intestinal regulatory T-cells (Treg cells). Here, we assessed the effect of SIN on the generation and function of Treg cells in autoimmune arthritis, and the underlying mechanisms in view of aryl hydrocarbon receptor (AhR). The proportions of Treg cells and IL-17-producing T-cells (Th17 cells) differentiated from naive T-cells were analyzed by flow cytometric analysis. The AhR agonistic effect of SIN was tested by analyzing the activation of downstream signaling pathways and target genes. The dependence of intestinal Treg cell induction and arthritis alleviation by SIN on AhR activation was confirmed in a mouse collagen-induced arthritis (CIA) model. SIN promoted the differentiation and function of intestinal Treg cells in vitro. It induced the expression and activity of AhR target gene, promoted AhR/Hsp90 dissociation and AhR nuclear translocation, induced XRE reporter activity, and facilitated AhR/XRE binding in vitro, displaying the potential to be an agonist of AhR. In CIA mice, SIN induced the generation of intestinal Treg cells, and facilitated the immunosuppressive function of these Treg cells as shown by an adoptive transfer test. In addition, the induction of intestinal Treg cells and the anti-arthritic effect of SIN in CIA mice could be largely diminished by the AhR antagonist resveratrol. SIN attenuates arthritis by promoting the generation and function of Treg cells in an AhR-dependent manner.

  15. Polyphyllin G induce apoptosis and autophagy in human nasopharyngeal cancer cells by modulation of AKT and mitogen-activated protein kinase pathways in vitro and in vivo.

    PubMed

    Chen, Jui-Chieh; Hsieh, Ming-Ju; Chen, Chih-Jung; Lin, Jen-Tsun; Lo, Yu-Sheng; Chuang, Yi-Ching; Chien, Su-Yu; Chen, Mu-Kuan

    2016-10-25

    Polyphyllin G (also call polyphyllin VII), extract from rhizomes of Paris yunnanensis Franch, has been demonstrated to have strong anticancer activities in a wide variety of human cancer cell lines. Previous studies found that Polyphyllin G induced apoptotic cell death in human hepatoblastoma cancer and lung cancer cells. However, the underlying mechanisms of autophagy in human nasopharyngeal carcinoma (NPC) remain unclear. In this study, Polyphyllin G can potently induced apoptosis dependent on the activations of caspase-8, -3, and -9 and the changes of Bcl-2, Bcl-xL and Bax protein expression in different human NPC cell lines (HONE-1 and NPC-039). The amount of both LC3-II and Beclin-1 was intriguingly increased suggest that autophagy was induced in Polyphyllin G-treated NPC cells. To further clarify whether Polyphyllin G-induced apoptosis and autophagy depended on AKT/ERK/JNK/p38 MAPK signaling pathways, cells were combined treated with AKT inhibitor (LY294002), ERK1/2 inhibitor (U0126), p38 MAPK inhibitor (SB203580), or JNK inhibitor (SP600125). These results demonstrated that Polyphyllin G induced apoptosis in NPC cells through activation of ERK, while AKT, p38 MAPK and JNK were responsible for Polyphyllin G-induced autophagy. Finally, an administration of Polyphyllin G effectively suppressed the tumor growth in the NPC carcinoma xenograft model in vivo. In conclusion, our results reveal that Polyphyllin G inhibits cell viability and induces apoptosis and autophagy in NPC cancer cells, suggesting that Polyphyllin G is an attractive candidate for tumor therapies. Polyphyllin G may promise candidate for development of antitumor drugs targeting nasopharyngeal carcinoma.

  16. Polyphyllin G induce apoptosis and autophagy in human nasopharyngeal cancer cells by modulation of AKT and mitogen-activated protein kinase pathways in vitro and in vivo

    PubMed Central

    Chen, Chih-Jung; Lin, Jen-Tsun; Lo, Yu-Sheng; Chuang, Yi-Ching; Chien, Su-Yu; Chen, Mu-Kuan

    2016-01-01

    Polyphyllin G (also call polyphyllin VII), extract from rhizomes of Paris yunnanensis Franch, has been demonstrated to have strong anticancer activities in a wide variety of human cancer cell lines. Previous studies found that Polyphyllin G induced apoptotic cell death in human hepatoblastoma cancer and lung cancer cells. However, the underlying mechanisms of autophagy in human nasopharyngeal carcinoma (NPC) remain unclear. In this study, Polyphyllin G can potently induced apoptosis dependent on the activations of caspase-8, -3, and -9 and the changes of Bcl-2, Bcl-xL and Bax protein expression in different human NPC cell lines (HONE-1 and NPC-039). The amount of both LC3-II and Beclin-1 was intriguingly increased suggest that autophagy was induced in Polyphyllin G-treated NPC cells. To further clarify whether Polyphyllin G-induced apoptosis and autophagy depended on AKT/ERK/JNK/p38 MAPK signaling pathways, cells were combined treated with AKT inhibitor (LY294002), ERK1/2 inhibitor (U0126), p38 MAPK inhibitor (SB203580), or JNK inhibitor (SP600125). These results demonstrated that Polyphyllin G induced apoptosis in NPC cells through activation of ERK, while AKT, p38 MAPK and JNK were responsible for Polyphyllin G-induced autophagy. Finally, an administration of Polyphyllin G effectively suppressed the tumor growth in the NPC carcinoma xenograft model in vivo. In conclusion, our results reveal that Polyphyllin G inhibits cell viability and induces apoptosis and autophagy in NPC cancer cells, suggesting that Polyphyllin G is an attractive candidate for tumor therapies. Polyphyllin G may promise candidate for development of antitumor drugs targeting nasopharyngeal carcinoma. PMID:27602962

  17. Mitomycin C-DNA adducts generated by DT-diaphorase. Revised mechanism of the enzymatic reductive activation of mitomycin C.

    PubMed

    Suresh Kumar, G; Lipman, R; Cummings, J; Tomasz, M

    1997-11-18

    Mitomycin C (MC) was reductively activated by DT-diaphorase [DTD; NAD(P)H:quinone oxidoreductase] from rat liver carcinoma cells in the presence of Micrococcus lysodeicticus DNA at pH 5.8 and 7.4. The resulting alkylated MC-DNA complexes were digested to the nucleoside level and the covalent MC-nucleoside adducts were separated, identified, and quantitatively analyzed by HPLC. In analogous experiments, two other flavoreductases, NADH-cytochrome c reductase and NADPH-cytochrome c reductase, as well as two chemical reductive activating agents Na2S2O4 and H2/PtO2 were employed as activators for the alkylation of DNA by MC. DTD as well as all the other activators generated the four known major guanine-N2-MC adducts at both pHs. In addition, at the lower pH, the guanine-N7-linked adducts of 2,7-diaminomitosene were detectable in the adduct patterns. At a given pH all the enzymatic and chemical reducing agents generated very similar adduct patterns which, however, differed dramatically at the acidic as compared to the neutral pH. Overall yield of MC adducts was 3-4-fold greater at pH 7.4 than at 5. 8 except in the case of DTD when it was 4-fold lower. Without exception, however, cross-link adduct yields were greater at the acidic pH (2-10-fold within the series). The ratio of adducts of bifunctional activation to those of monofunctional activation was 6-20-fold higher at the acidic as compared to the neutral pH. A comprehensive mechanism of the alkylation of DNA by activated MC was derived from the DNA adduct analysis which complements earlier model studies of the activation of MC. The mechanism consists of three competing activation pathways yielding three different DNA-reactive electrophiles 11, 12, and 17 which generate three unique sets of DNA adducts as endproducts. The relative amounts of these adducts are diagnostic of the relative rates of the competing pathways in vitro, and most likely, in vivo. Factors that influence the relative rates of individual pathways

  18. 3D active shape models of human brain structures: application to patient-specific mesh generation

    NASA Astrophysics Data System (ADS)

    Ravikumar, Nishant; Castro-Mateos, Isaac; Pozo, Jose M.; Frangi, Alejandro F.; Taylor, Zeike A.

    2015-03-01

    The use of biomechanics-based numerical simulations has attracted growing interest in recent years for computer-aided diagnosis and treatment planning. With this in mind, a method for automatic mesh generation of brain structures of interest, using statistical models of shape (SSM) and appearance (SAM), for personalised computational modelling is presented. SSMs are constructed as point distribution models (PDMs) while SAMs are trained using intensity profiles sampled from a training set of T1-weighted magnetic resonance images. The brain structures of interest are, the cortical surface (cerebrum, cerebellum & brainstem), lateral ventricles and falx-cerebri membrane. Two methods for establishing correspondences across the training set of shapes are investigated and compared (based on SSM quality): the Coherent Point Drift (CPD) point-set registration method and B-spline mesh-to-mesh registration method. The MNI-305 (Montreal Neurological Institute) average brain atlas is used to generate the template mesh, which is deformed and registered to each training case, to establish correspondence over the training set of shapes. 18 healthy patients' T1-weightedMRimages form the training set used to generate the SSM and SAM. Both model-training and model-fitting are performed over multiple brain structures simultaneously. Compactness and generalisation errors of the BSpline-SSM and CPD-SSM are evaluated and used to quantitatively compare the SSMs. Leave-one-out cross validation is used to evaluate SSM quality in terms of these measures. The mesh-based SSM is found to generalise better and is more compact, relative to the CPD-based SSM. Quality of the best-fit model instance from the trained SSMs, to test cases are evaluated using the Hausdorff distance (HD) and mean absolute surface distance (MASD) metrics.

  19. Generation of Xenotransplants from Human Cancer Biopsies to Assess Anti-cancer Activities of HDACi.

    PubMed

    Mullins, Christina Susanne; Bock, Stephanie; Krohn, Mathias; Linnebacher, Michael

    2017-01-01

    Human tumor in vivo cancer models raised in immunodeficient mice, the so-called patient-derived xenografts, are increasingly in use in preclinical development and evaluation of novel drug candidates including HDAC inhibitors. Here, we describe the techniques needed to generate novel patient-derived xenografts. The focus lies on vitally frozen tumor biopsies as starting material. First, the preparative steps on the animals, followed by the engraftment procedure itself, the tumor growth surveillance, the explantation procedure, and finally the handling of obtained xenograft tissues are described step by step. This technical description is completed by numerous tips and alternatives designed to allow for an easy adaptation and transfer to other laboratories.

  20. Preparing the periphery for a subsequent behavior: motor neuronal activity during biting generates little force but prepares a retractor muscle to generate larger forces during swallowing in Aplysia.

    PubMed

    Lu, Hui; McManus, Jeffrey M; Cullins, Miranda J; Chiel, Hillel J

    2015-03-25

    Some behaviors occur in obligatory sequence, such as reaching before grasping an object. Can the earlier behavior serve to prepare the musculature for the later behavior? If it does, what is the underlying neural mechanism of the preparation? To address this question, we examined two feeding behaviors in the marine mollusk Aplysia californica, one of which must precede the second: biting and swallowing. Biting is an attempt to grasp food. When that attempt is successful, the animal immediately switches to swallowing to ingest food. The main muscle responsible for pulling food into the buccal cavity during swallowing is the I3 muscle, whose motor neurons B6, B9, and B3 have been previously identified. By performing recordings from these neurons in vivo in intact, behaving animals or in vitro in a suspended buccal mass preparation, we demonstrated that the frequencies and durations of these motor neurons increased from biting to swallowing. Using the physiological patterns of activation to drive these neurons intracellularly, we further demonstrated that activating them using biting-like frequencies and durations, either alone or in combination, generated little or no force in the I3 muscle. When biting-like patterns preceded swallowing-like patterns, however, the forces during the subsequent swallowing-like patterns were significantly enhanced. Sequences of swallowing-like patterns, either with these neurons alone or in combination, further enhanced forces in the I3 muscle. These results suggest a novel mechanism for enhancing force production in a muscle, and may be relevant to understanding motor control in vertebrates.

  1. Dosimetric difference amongst 3 techniques: TomoTherapy, sliding-window intensity-modulated radiotherapy (IMRT), and RapidArc radiotherapy in the treatment of late-stage nasopharyngeal carcinoma (NPC)

    SciTech Connect

    Lee, Francis Kar-ho Yip, Celia Wai-yi; Cheung, Frankie Chun-hung; Leung, Alex Kwok-cheung; Chau, Ricky Ming-chun; Ngan, Roger Kai-cheong

    2014-04-01

    To investigate the dosimetric difference amongst TomoTherapy, sliding-window intensity-modulated radiotherapy (IMRT), and RapidArc radiotherapy in the treatment of late-stage nasopharyngeal carcinoma (NPC). Ten patients with late-stage (Stage III or IV) NPC treated with TomoTherapy or IMRT were selected for the study. Treatment plans with these 3 techniques were devised according to departmental protocol. Dosimetric parameters for organ at risk and treatment targets were compared between TomoTherapy and IMRT, TomoTherapy and RapidArc, and IMRT and RapidArc. Comparison amongst the techniques was done by statistical tests on the dosimetric parameters, total monitor unit (MU), and expected delivery time. All 3 techniques achieved similar target dose coverage. TomoTherapy achieved significantly lower doses in lens and mandible amongst the techniques. It also achieved significantly better dose conformity to the treatment targets. RapidArc achieved significantly lower dose to the eye and normal tissue, lower total MU, and less delivery time. The dosimetric advantages of the 3 techniques were identified in the treatment of late-stage NPC. This may serve as a guideline for selection of the proper technique for different clinical cases.

  2. Mechanical stimulation of skeletal muscle generates lipid-related second messengers by phospholipase activation

    NASA Technical Reports Server (NTRS)

    Vandenburgh, H. H.; Shansky, J.; Karlisch, P.; Solerssi, R. L.

    1993-01-01

    Repetitive mechanical stimulation of cultured avian skeletal muscle increases the synthesis of prostaglandins (PG) E2 and F2 alpha which regulate protein turnover rates and muscle cell growth. These stretch-induced PG increases are reduced in low extracellular calcium medium and by specific phospholipase inhibitors. Mechanical stimulation increases the breakdown rate of 3H-arachidonic acid labelled phospholipids, releasing free 3H-arachidonic acid, the rate-limiting precursor of PG synthesis. Mechanical stimulation also increases 3H-arachidonic acid labelled diacylglycerol formation and intracellular levels of inositol phosphates from myo-[2-3H]inositol labelled phospholipids. Phospholipase A2 (PLA2), phosphatidylinositol-specific phospholipase C (PLC), and phospholipase D (PLD) are all activated by stretch. The stretch-induced increases in PG production, 3H-arachidonic acid labelled phospholipid breakdown, and 3H-arachidonic acid labelled diacylglycerol formation occur independently of cellular electrical activity (tetrodotoxin insensitive) whereas the formation of inositol phosphates from myo-[2-3H]inositol labelled phospholipids is dependent on cellular electrical activity. These results indicate that mechanical stimulation increases the lipid-related second messengers arachidonic acid, diacylglycerol, and PG through activation of specific phospholipases such as PLA2 and PLD, but not by activation of phosphatidylinositol-specific PLC.

  3. Isomorphous substitution and the generation of catalytic activity in VPI-5

    SciTech Connect

    Kraushaar-Czarnetzki, B.; Dogterom, R.J.; Stork, W.H.J.; Emeis, K.A.; Van Braam Houckgeest, J.P. ) )

    1993-05-01

    VPI-5 and substituted derivatives containing silicon or magnesium have been synthesized, analyzed, and tested for their catalytic activity. By means of [sup 29]Si solid-state NMR, it could be shown that SAPO-VPI-5 prepared with polyphosphoric acid as a phosphorus source exhibits both [open quotes]silica patches[close quotes] and isolated silicon atoms incorporated in the framework. Broensted acid sites in SAPO-and MAPO-VPI-5 could be detected by means of infrared spectroscopy in combination with pyridine adsorption. The isomorphously substituted VPI-5 samples were active in the hydroconversion of n-heptane, giving both cracking and isomerization products. The highest activity and the highest yield in iso-heptane, however, were observed with AlPO-VPI-5 as a catalyst. The catalytic activity could be ascribed to the presence of unreacted alumina. Upon treatment with an EDTA solution the Al/P atomic ratio decreased from 1.29 to 1.07, and the purified AlPO-VPI-5 showed considerably reduced catalytic activity. 28 refs., 6 figs., 2 tabs.

  4. Mechanical stimulation of skeletal muscle generates lipid-related second messengers by phospholipase activation

    NASA Technical Reports Server (NTRS)

    Vandenburgh, Herman H.; Shansky, Janet; Karlisch, Patricia; Solerssi, Rosa Lopez

    1991-01-01

    Repetitive mechanical stimulation of cultured avian skeletal muscle increases the synthesis of prostaglandins E2 and F2(alpha) which regulate protein turnover rates and muscle cell growth. Mechnical stimulation significantly increases the breakdown rate of (3)H-arachidonic acid labelled phospholipids, releasing free (3)H-arachidonic acid, and the rate-limiting precursor of prostaglandin synthesis. Mechanical stimulation also significantly increases (3)H-arachidonic acid labelled diacylglycerol formation and intracellular levels of inositol phosphates from myo-2-(3)H inositol labelled phospholipids. Phospholipase A2, phosphatidylinositol-specific phospholipase C (PLC), and phospholipase D (PLD) are activated by stretch. The lipase inhibitors bromophenacylbromide and RHC80267 together reduce stretch-induced prostaglandin production by 73-83 percent. The stretch-induced increases in prostaglandin production, (3)H-arachidonic acid labelled phospholipid breakdown, and (3)H-arachidonic acid labelled diacylglycerol formation occur independently of cellular electrical activity (tetrodotoxin insensitive) whereas the formation of inositol phosphates from myo-2-(3)H inositol labelled phospholipids are dependent on cellular electrical activity. These results indicate that mechanical stimulation increases the lipid-related second messengers arachidonic acid, diacylglycerol, and prostaglandins through activation of specific phospholipases such as PLA2 and PLD, but not by activation of phosphatidylinositol-specific PLC.

  5. Evaluation of Antibacterial Activity and Reactive Species Generation of N-Benzenesulfonyl Derivatives of Heterocycles.

    PubMed

    Martínez, Sol Romina; Miana, Gisele Emilse; Albesa, Inés; Mazzieri, María Rosa; Becerra, María Cecilia

    2016-01-01

    Two N-benzenesulfonyl (BS) derivatives of 1,2,3,4-tetrahydroquinoline (THQ) were designed, prepared, and screened for antibacterial activity. This approach was based on combining the two privileged structures, BS and THQ, which are known to be active. The objective of this study was to evaluate the antibacterial activity of BS-THQ and its analogue 4-NH2BS-THQ, and to investigate the roles of reactive oxygen species and reactive nitrogen species in their lethality. Both showed bactericidal activity against Staphylococcus aureus ATCC 29213 and methicillin-resistant S. aureus (MRSA) ATCC 43300, with transmission electron microscopy revealing a disturbed membrane architecture. Furthermore, an increase of reactive oxygen species (ROS) in strains treated with BS-THQ with respect to the control was detected when fluorescent microscopy and spectrophotometric techniques were used. The analogue 4-NH2BS-THQ demonstrated a broader spectrum of activity than BS-THQ, with a minimum inhibitory concentration of 100 µg/mL against reference strains of S. aureus, Escherichia coli and Pseudomonas aeruginosa. The assayed compounds represent promising structures for the development of new synthetic classes of antimicrobials.

  6. Chronic NF-κB activation links COPD and lung cancer through generation of an immunosuppressive microenvironment in the lungs

    PubMed Central

    Zaynagetdinov, Rinat; Sherrill, Taylor P.; Gleaves, Linda A.; Hunt, Pierre; Han, Wei; McLoed, Allyson G.; Saxon, Jamie A.; Tanjore, Harikrishna; Gulleman, Peter M.; Young, Lisa R.; Blackwell, Timothy S.

    2016-01-01

    Nuclear Factor (NF)-κB is positioned to provide the interface between COPD and carcinogenesis through regulation of chronic inflammation in the lungs. Using a tetracycline-inducible transgenic mouse model that conditionally expresses activated IκB kinase β (IKKβ) in airway epithelium (IKTA), we found that sustained NF-κB signaling results in chronic inflammation and emphysema by 4 months. By 11 months of transgene activation, IKTA mice develop lung adenomas. Investigation of lung inflammation in IKTA mice revealed a substantial increase in M2-polarized macrophages and CD4+/CD25+/FoxP3+ regulatory T lymphocytes (Tregs). Depletion of alveolar macrophages in IKTA mice reduced Tregs, increased lung CD8+ lymphocytes, and reduced tumor numbers following treatment with the carcinogen urethane. Alveolar macrophages from IKTA mice supported increased generation of inducible Foxp3+ Tregs ex vivo through expression of TGFβ and IL-10. Targeting of TGFβ and IL-10 reduced the ability of alveolar macrophages from IKTA mice to induce Foxp3 expression on T cells. These studies indicate that sustained activation of NF-κB pathway links COPD and lung cancer through generation and maintenance of a pro-tumorigenic inflammatory environment consisting of alternatively activated macrophages and regulatory T cells. PMID:26756215

  7. Chronic NF-κB activation links COPD and lung cancer through generation of an immunosuppressive microenvironment in the lungs.

    PubMed

    Zaynagetdinov, Rinat; Sherrill, Taylor P; Gleaves, Linda A; Hunt, Pierre; Han, Wei; McLoed, Allyson G; Saxon, Jamie A; Tanjore, Harikrishna; Gulleman, Peter M; Young, Lisa R; Blackwell, Timothy S

    2016-02-02

    Nuclear Factor (NF)-κB is positioned to provide the interface between COPD and carcinogenesis through regulation of chronic inflammation in the lungs. Using a tetracycline-inducible transgenic mouse model that conditionally expresses activated IκB kinase β (IKKβ) in airway epithelium (IKTA), we found that sustained NF-κB signaling results in chronic inflammation and emphysema by 4 months. By 11 months of transgene activation, IKTA mice develop lung adenomas. Investigation of lung inflammation in IKTA mice revealed a substantial increase in M2-polarized macrophages and CD4+/CD25+/FoxP3+ regulatory T lymphocytes (Tregs). Depletion of alveolar macrophages in IKTA mice reduced Tregs, increased lung CD8+ lymphocytes, and reduced tumor numbers following treatment with the carcinogen urethane. Alveolar macrophages from IKTA mice supported increased generation of inducible Foxp3+ Tregs ex vivo through expression of TGFβ and IL-10. Targeting of TGFβ and IL-10 reduced the ability of alveolar macrophages from IKTA mice to induce Foxp3 expression on T cells. These studies indicate that sustained activation of NF-κB pathway links COPD and lung cancer through generation and maintenance of a pro-tumorigenic inflammatory environment consisting of alternatively activated macrophages and regulatory T cells.

  8. Generation of spectral clusters in a mixture of noble and Raman-active gases.

    PubMed

    Hosseini, Pooria; Abdolvand, Amir; St J Russell, Philip

    2016-12-01

    We report a novel scheme for the generation of dense clusters of Raman sidebands. The scheme uses a broadband-guiding hollow-core photonic crystal fiber (HC-PCF) filled with a mixture of H2, D2, and Xe for efficient interaction between the gas mixture and a green laser pump pulse (532 nm, 1 ns) of only 5 μJ of energy. This results in the generation from noise of more than 135 rovibrational Raman sidebands covering the visible spectral region with an average spacing of only 2.2 THz. Such a spectrally dense and compact fiber-based source is ideal for applications where closely spaced narrow-band laser lines with high spectral power density are required, such as in spectroscopy and sensing. When the HC-PCF is filled with a H2-D2 mixture, the Raman comb spans the spectral region from the deep UV (280 nm) to the near infrared (1000 nm).

  9. Role of turnover in active stress generation in a filament network

    PubMed Central

    Hiraiwa, Tetsuya; Salbreux, Guillaume

    2016-01-01

    We study the effect of turnover of cross-linkers, motors, and filaments on the generation of a contractile stress in a network of filaments connected by passive cross-linkers and subjected to the forces exerted by molecular motors. We perform numerical simulations where filaments are treated as rigid rods and molecular motors move fast compared to the time scale of an exchange of cross-linkers. We show that molecular motors create a contractile stress above a critical number of cross-linkers. When passive cross-linkers are allowed to turn over, the stress exerted by the network vanishes due to the formation of clusters. When both filaments and passive cross-linkers turn over, clustering is prevented and the network reaches a dynamic contractile steady state. A maximum stress is reached for an optimum ratio of the filament and cross-linker turnover rates. Taken together, our work reveals conditions for stress generation by molecular motors in a fluid isotropic network of rearranging filaments. PMID:27203344

  10. Role of Turnover in Active Stress Generation in a Filament Network

    NASA Astrophysics Data System (ADS)

    Hiraiwa, Tetsuya; Salbreux, Guillaume

    2016-05-01

    We study the effect of turnover of cross-linkers, motors, and filaments on the generation of a contractile stress in a network of filaments connected by passive cross-linkers and subjected to the forces exerted by molecular motors. We perform numerical simulations where filaments are treated as rigid rods and molecular motors move fast compared to the time scale of an exchange of cross-linkers. We show that molecular motors create a contractile stress above a critical number of cross-linkers. When passive cross-linkers are allowed to turn over, the stress exerted by the network vanishes due to the formation of clusters. When both filaments and passive cross-linkers turn over, clustering is prevented and the network reaches a dynamic contractile steady state. A maximum stress is reached for an optimum ratio of the filament and cross-linker turnover rates. Taken together, our work reveals conditions for stress generation by molecular motors in a fluid isotropic network of rearranging filaments.

  11. Mreg Activity in Tumor Response to Photodynamic Therapy and Photodynamic Therapy-Generated Cancer Vaccines

    PubMed Central

    Korbelik, Mladen; Banáth, Judith; Zhang, Wei

    2016-01-01

    Myeloid regulatory cells (Mregs) are, together with regulatory T cells (Tregs), a dominant effector population responsible for restriction of the duration and strength of antitumor immune response. Photodynamic therapy (PDT) and cancer vaccines generated by PDT are modalities whose effectiveness in tumor destruction is closely dependent on the associated antitumor immune response. The present study investigated whether the immunodepletion of granulocytic Mregs in host mice by anti-GR1 antibody would improve the response of tumors to PDT or PDT vaccines in these animals. Anti-GR1 administration immediately after Temoporfin-PDT of mouse SCCVII tumors abrogated curative effect of PDT. The opposite effect, increasing PDT-mediated tumor cure-rates was attained by delaying anti-GR1 treatment to 1 h post PDT. With PDT vaccines, multiple anti-GR1 administrations (days 0, 4, and 8 post vaccination) improved the therapy response with SCCVII tumors. The results with PDT suggest that neutrophils (boosting antitumor effect of this therapy) that are engaged immediately after photodynamic light treatment are within one hour replaced with a different myeloid population, presumably Mregs that hampers the therapy-mediated antitumor effect. Anti-GR1 antibody, when used with optimal timing, can improve the efficacy of both PDT of tumors in situ and PDT-generated cancer vaccines. PMID:27754452

  12. What is the importance of abnormal "background" activity in seizure generation?

    PubMed

    Staba, Richard J; Worrell, Gregory A

    2014-01-01

    Investigations of interictal epileptiform spikes and seizures have played a central role in the study of epilepsy. The background EEG activity, however, has received less attention. In this chapter we discuss the characteristic features of the background activity of the brain when individuals are at rest and awake (resting wake) and during sleep. The characteristic rhythms of the background EEG are presented, and the presence of 1/f (β) behavior of the EEG power spectral density is discussed and its possible origin and functional significance. The interictal EEG findings of focal epilepsy and the impact of interictal epileptiform spikes on cognition are also discussed.

  13. Degree of conversion of nanofilled and microhybrid composite resins photo-activated by different generations of LEDs

    PubMed Central

    RIBEIRO, Benicia Carolina Iaskieviscz; BOAVENTURA, Juliana Maria Capelozza; de BRITO-GONÇALVES, Joel; RASTELLI, Alessandra Nara de Souza; BAGNATO, Vanderlei Salvador; SAAD, José Roberto Cury

    2012-01-01

    Objective This study aimed at evaluating the degree of conversion (DC) of four composite resins, being one nanofilled and 3 microhybrid resins, photo-activated with second- and third-generation light-emitting diodes (LEDs). Material and methods FiltekTM Z350 nanofilled composite resins and Amelogen® Plus, Vit-l-escenceTM and Opallis microhybrid resins were photo-activated with two second-generation LEDs (Radii-cal and Elipar Free LightTM 2) and one third-generation LED (Ultra-Lume LED 5) by continuous light mode, and a quartz halogen-tungsten bulb (QHT, control). After 24 h of storage, the samples were pulverized into fine powder and 5 mg of each material were mixed with 100 mg of potassium bromide (KBr). After homogenization, they were pressed, which resulted in a pellet that was evaluated using an infrared spectromer (Nexus 470, Thermo Nicolet) equipped with TGS detector using diffuse reflectance (32 scans, resolution of 4 cm-1) coupled to a computer. The percentage of unreacted carbon-carbon double bonds (% C=C) was determined from the ratio of absorbance intensities of aliphatic C=C (peak at 1637 cm-1) against internal standard before and after curing of the specimen: aromatic C-C (peak at 1610 cm-1). Results The ANOVA showed a significant effect on the interaction between the light-curing units (LCUs) and the composite resins (p<0.001). The Tukey's test showed that the nanofilled resin (FiltekTM Z350) and Opallis when photo-activated by the halogen lamp (QTH) had the lowest DC compared with the other microhybrid composite resins. The DC of the nanofilled resin (FiltekTM Z350) was also lower using LEDs. The highest degrees of conversion were obtained using the third-generation LED and one of second-generation LEDs (Elipar Free LightTM 2). Conclusions The nanofilled resin showed the lowest DC, and the Vit-l-escenceTM microhybrid composite resin showed the highest DC. Among the LCUs, it was not possible to establish an order, even though the second-generation

  14. Elaboration of copper-oxygen mediated C-H activation chemistry in consideration of future fuel and feedstock generation.

    PubMed

    Lee, Jung Yoon; Karlin, Kenneth D

    2015-04-01

    To contribute solutions to current energy concerns, improvements in the efficiency of dioxygen mediated C-H bond cleavage chemistry, for example, selective oxidation of methane to methanol, could minimize losses in natural gas usage or produce feedstocks for fuels. Oxidative C-H activation is also a component of polysaccharide degradation, potentially affording alternative biofuels from abundant biomass. Thus, an understanding of active-site chemistry in copper monooxygenases, those activating strong C-H bonds is briefly reviewed. Then, recent advances in the synthesis-generation and study of various copper-oxygen intermediates are highlighted. Of special interest are cupric-superoxide, Cu-hydroperoxo and Cu-oxy complexes. Such investigations can contribute to an enhanced future application of C-H oxidation or oxygenation processes using air, as concerning societal energy goals.

  15. Elaboration of Copper-Oxygen Mediated C–H Activation Chemistry in Consideration of Future Fuel and Feedstock Generation

    PubMed Central

    Lee, Jung Yoon; Karlin, Kenneth D

    2015-01-01

    To contribute solutions for current energy concerns, improvements in the efficiency of C-H bond cleavage chemistry, e.g., selective oxidation of methane to methanol, could minimize losses in natural gas usage or produce feedstocks for fuels. Oxidative C-H activation is also a component of polysaccharide degradation, affording alternative biofuels from abundant biomass. Thus, an understanding of active-site chemistry in copper monooxygenases, those activating strong C-H bonds is briefly reviewed. Then, recent advances in the synthesis-generation and study of various copper-oxygen intermediates are highlighted. Of special interest are cupric-superoxide, Cu-hydroperoxo and Cu-oxy complexes. Such investigations can contribute to an enhanced future application of C-H oxidation or oxygenation processes using air, as concerning societal energy goals. PMID:25756327

  16. Approaches for the generation of active papain-like cysteine proteases from inclusion bodies of Escherichia coli.

    PubMed

    Ling, Chunfang; Zhang, Junyan; Lin, Deqiu; Tao, Ailin

    2015-05-01

    Papain-like cysteine proteases are widely expressed, fulfill specific functions in extracellular matrix turnover, antigen presentation and processing events, and may represent viable drug targets for major diseases. In depth and rigorous studies of the potential for these proteins to be targets for drug development require sufficient amounts of protease protein that can be used for both experimental and therapeutic purposes. Escherichia coli was widely used to express papain-like cysteine proteases, but most of those proteases are produced in insoluble inclusion bodies that need solubilizing, refolding, purifying and activating. Refolding is the most critical step in the process of generating active cysteine proteases and the current approaches to refolding include dialysis, dilution and chromatography. Purification is mainly achieved by various column chromatography. Finally, the attained refolded proteases are examined regarding their protease structures and activities.

  17. Optoacoustic tweezers: a programmable, localized cell concentrator based on opto-thermally generated, acoustically activated, surface bubbles

    PubMed Central

    Zhao, Yanhui; Li, Sixing; Rufo, Joseph; Yang, Shikuan; Guo, Feng; Huang, Tony Jun

    2014-01-01

    We present a programmable, biocompatible technique for dynamically concentrating and patterning particles and cells in a microfluidic device. Since our technique utilizes opto-thermally generated, acoustically activated, surface bubbles, we name it “optoacoustic tweezers.” The optoacoustic tweezers are capable of concentrating particles/cells at any prescribed locations in a microfluidic chamber without the use of permanent structures, rendering it particularly useful for the formation of flexible, complex cell patterns. Additionally, this technique has demonstrated excellent biocompatibility and can be conveniently integrated with other microfluidic units. In our experiments, micro-bubbles were generated by focusing a 405 nm diode laser onto a gold-coated glass chamber. By properly tuning the laser, we demonstrate precise control over the position and size of the generated bubbles. Acoustic waves were then applied to activate the surface bubbles, causing them to oscillate at an optimized frequency. The resulting acoustic radiation force allowed us to locally trap particles/cells, including 15 μm polystyrene beads and HeLa cells, around each bubble. Cell-adhesion tests were also conducted after cell concentrating to confirm the biocompatibility of this technique. PMID:23511348

  18. Steps in reductive activation of the disulfide-generating enzyme Ero1p

    PubMed Central

    Heldman, Nimrod; Vonshak, Ohad; Sevier, Carolyn S; Vitu, Elvira; Mehlman, Tevie; Fass, Deborah

    2010-01-01

    Ero1p is the primary catalyst of disulfide bond formation in the yeast endoplasmic reticulum (ER). Ero1p contains a pair of essential disulfide bonds that participate directly in the electron transfer pathway from substrate thiol groups to oxygen. Remarkably, elimination of certain other Ero1p disulfides by mutation enhances enzyme activity. In particular, the C150A/C295A Ero1p mutant exhibits increased thiol oxidation in vitro and in vivo and interferes with redox homeostasis in yeast cells by hyperoxidizing the ER. Inhibitory disulfides of Ero1p are thus important for enzyme regulation. To visualize the differences between de-regulated and wild-type Ero1p, we determined the crystal structure of Ero1p C150A/C295A. The structure revealed local changes compared to the wild-type enzyme around the sites of mutation, but no conformational transitions within 25 Å of the active site were observed. To determine how the C150—C295 disulfide nonetheless participates in redox regulation of Ero1p, we analyzed using mass spectrometry the changes in Ero1p disulfide connectivity as a function of time after encounter with reducing substrates. We found that the C150—C295 disulfide sets a physiologically appropriate threshold for enzyme activation by guarding a key neighboring disulfide from reduction. This study illustrates the diverse and interconnected roles that disulfides can play in redox regulation of protein activity. PMID:20669236

  19. Water contaminated with Didymosphenia geminata generates changes in Salmo salar spermatozoa activation times.

    PubMed

    Olivares, Pamela; Orellana, Paola; Guerra, Guillermo; Peredo-Parada, Matías; Chavez, Viviana; Ramirez, Alfredo; Parodi, Jorge

    2015-06-01

    Didimosphenia geminata ("didymo"), has become a powerful and devastating river plague in Chile. A system was developed in D. geminata channels with the purpose evaluating the effects of water polluted with didymo on the activation of Atlantic salmon (Salmo salar) spermatozoa. Results indicate that semen, when activated with uncontaminated river water had an average time of 60±21s. When using Powermilt, (a commercial activator), times of 240±21s are achieved, while rivers contaminated with D. geminata achieve a motility time of 30±12s. Interestingly enough, the kinetic parameters of VSL, VCL and VAP showed no significant changes under all of the conditions. Furthermore, the presence of D. geminata reduces activation time of the samples as the cells age, indicating increased effects in spermatozoa that are conserved for more than 5 days. D. geminata has antioxidant content, represented by polyphenols; 200ppm of polyphenol were obtained in this study per 10g of microalgae. Spermatozoa exposed to these extracts showed a reduction in mobility time in a dose dependent manner, showing an IC50 of 15ppm. The results suggest an effect on spermatozoa activation, possibly due to the release of polyphenols present in contaminated rivers, facilitating the alteration of sperm motility times, without affecting the viability or kinetics of the cells. These findings have important implications for current policy regarding the control of the algae. Current control measures focus on the number of visible species, and not on the compounds that they release, which this study shows, also have a problematic effect on salmon production.

  20. Thermo-Magneto-Electric Generator Arrays for Active Heat Recovery System

    PubMed Central

    Chun, Jinsung; Song, Hyun-Cheol; Kang, Min-Gyu; Kang, Han Byul; Kishore, Ravi Anant; Priya, Shashank

    2017-01-01

    Continued emphasis on development of thermal cooling systems is being placed that can cycle low grade heat. Examples include solar powered unmanned aerial vehicles (UAVs) and data storage servers. The power efficiency of solar module degrades at elevated temperature, thereby, necessitating the need for heat extraction system. Similarly, data centres in wireless computing system are facing increasing efficiency challenges due to high power consumption associated with managing the waste heat. We provide breakthrough in addressing these problems by developing thermo-magneto-electric generator (TMEG) arrays, composed of soft magnet and piezoelectric polyvinylidene difluoride (PVDF) cantilever. TMEG can serve dual role of extracting the waste heat and converting it into useable electricity. Near room temperature second-order magnetic phase transition in soft magnetic material, gadolinium, was employed to obtain mechanical vibrations on the PVDF cantilever under small thermal gradient. TMEGs were shown to achieve high vibration frequency at small temperature gradients, thereby, demonstrating effective heat transfer. PMID:28145516

  1. Thermo-Magneto-Electric Generator Arrays for Active Heat Recovery System.

    PubMed

    Chun, Jinsung; Song, Hyun-Cheol; Kang, Min-Gyu; Kang, Han Byul; Kishore, Ravi Anant; Priya, Shashank

    2017-02-01

    Continued emphasis on development of thermal cooling systems is being placed that can cycle low grade heat. Examples include solar powered unmanned aerial vehicles (UAVs) and data storage servers. The power efficiency of solar module degrades at elevated temperature, thereby, necessitating the need for heat extraction system. Similarly, data centres in wireless computing system are facing increasing efficiency challenges due to high power consumption associated with managing the waste heat. We provide breakthrough in addressing these problems by developing thermo-magneto-electric generator (TMEG) arrays, composed of soft magnet and piezoelectric polyvinylidene difluoride (PVDF) cantilever. TMEG can serve dual role of extracting the waste heat and converting it into useable electricity. Near room temperature second-order magnetic phase transition in soft magnetic material, gadolinium, was employed to obtain mechanical vibrations on the PVDF cantilever under small thermal gradient. TMEGs were shown to achieve high vibration frequency at small temperature gradients, thereby, demonstrating effective heat transfer.

  2. Thermo-Magneto-Electric Generator Arrays for Active Heat Recovery System

    NASA Astrophysics Data System (ADS)

    Chun, Jinsung; Song, Hyun-Cheol; Kang, Min-Gyu; Kang, Han Byul; Kishore, Ravi Anant; Priya, Shashank

    2017-02-01

    Continued emphasis on development of thermal cooling systems is being placed that can cycle low grade heat. Examples include solar powered unmanned aerial vehicles (UAVs) and data storage servers. The power efficiency of solar module degrades at elevated temperature, thereby, necessitating the need for heat extraction system. Similarly, data centres in wireless computing system are facing increasing efficiency challenges due to high power consumption associated with managing the waste heat. We provide breakthrough in addressing these problems by developing thermo-magneto-electric generator (TMEG) arrays, composed of soft magnet and piezoelectric polyvinylidene difluoride (PVDF) cantilever. TMEG can serve dual role of extracting the waste heat and converting it into useable electricity. Near room temperature second-order magnetic phase transition in soft magnetic material, gadolinium, was employed to obtain mechanical vibrations on the PVDF cantilever under small thermal gradient. TMEGs were shown to achieve high vibration frequency at small temperature gradients, thereby, demonstrating effective heat transfer.

  3. Comparison The Effects of Two Monocyte Isolation Methods, Plastic Adherence and Magnetic Activated Cell Sorting Methods, on Phagocytic Activity of Generated Dendritic Cells

    PubMed Central

    Delirezh, Nowruz; Shojaeefar, Ehsan; Parvin, Parva; Asadi, Behnaz

    2013-01-01

    Objective: It is believed that monocyte isolation methods and maturation factors affect the phenotypic and functional characteristics of resultant dendritic cells (DC). In the present study, we compared two monocyte isolation methods, including plastic adherence-dendritic cells (Adh-DC) and magnetic activated cell sorting- dendritic cells (MACS-DC), and their effects on phagocytic activity of differentiated immature DCs (immDCs). Materials and Methods: : In this experimental study, immDCs were generated from plastic adherence and MACS isolated monocytes in the presence of granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin 4 (IL-4) in five days. The phagocytic activity of immDCs was analyzed by fluorescein isothiocyanate (FITC)-conjugated latex bead using flow cytometry. One way ANOVA test was used for statistical analysis of differences among experimental groups, including Adh-DC and MACS-DC groups. Results: We found that phagocytic activity of Adh-DC was higher than MACS-DC, whereas the mean fluorescence intensity (MFI) of phagocytic cells was higher in MACS-DC (p<0.05). Conclusion: : We concluded that it would be important to consider phagocytosis parameters of generated DCs before making any decision about monocyte isolation methods to have fully functional DCs. PMID:24027662

  4. Generation of parthenogenetic goat blastocysts: effects of different activation methods and culture media.

    PubMed

    Malik, Hruda Nanda; Singhal, Dinesh Kumar; Saugandhika, Shrabani; Dubey, Amit; Mukherjee, Ayan; Singhal, Raxita; Kumar, Sudarshan; Kaushik, Jai Kumar; Mohanty, Ashok Kumar; Das, Bikash Chandra; Bag, Sadhan; Bhanja, Subrata Kumar; Malakar, Dhruba

    2015-06-01

    The present study was carried out to investigate the effects of different activation methods and culture media on the in vitro development of parthenogenetic goat blastocysts. Calcium (Ca2+) ionophore, ethanol or a combination of the two, used as activating reagents, and embryo development medium (EDM), modified Charles Rosenkrans (mCR2a) medium and research vitro cleave (RVCL) medium were used to evaluate the developmental competence of goat blastocysts. Quantitative expression of apoptosis, stress and developmental competence-related genes were analysed in different stages of embryos. In RVCL medium, the cleavage rate of Ca2+ ionophore-treated oocytes (79.61 ± 0.86) was significantly (P < 0.05) higher than in ethanol (74.90 ± 1.51) or in the combination of both Ca2+ ionophore and ethanol. In mCR2a or EDM, hatched blastocyst production rate of Ca2+ ionophore-treated oocytes (8.33 ± 1.44) was significantly higher than in ethanol (6.46 ± 0.11) or in the combined treatment (6.70 ± 0.24). In ethanol, the cleavage, blastocyst and hatched blastocyst production rates in RVCL medium (74.90 ± 1.51, 18.30 ± 1.52 and 8.24 ± 0.15, respectively) were significantly higher than in EDM (67.81 ± 3.21, 14.59 ± 0.27 and 5.59 ± 0.42) or mCR2a medium (65.09 ± 1.57, 15.36 ± 0.52 and 6.46 ± 0.11). The expression of BAX, Oct-4 and GlUT1 transcripts increased gradually from 2-cell stage to blastocyst-stage embryos, whereas the transcript levels of Bcl-2 and MnSOD were significantly lower in blastocysts. In addition, different activation methods and culture media had little effect on the pattern of variation and relative abundance of the above genes in different stages of parthenogenetic activated goat embryos. In conclusion, Ca2+ ionophore as the activating agent, and RVCL as the culture medium are better than other tested options for development of parthenogenetic activated goat blastocysts.

  5. Integrated Analysis of Contractile Kinetics, Force Generation, and Electrical Activity in Single Human Stem Cell-Derived Cardiomyocytes.

    PubMed

    Kijlstra, Jan David; Hu, Dongjian; Mittal, Nikhil; Kausel, Eduardo; van der Meer, Peter; Garakani, Arman; Domian, Ibrahim J

    2015-12-08

    The quantitative analysis of cardiomyocyte function is essential for stem cell-based approaches for the in vitro study of human cardiac physiology and pathophysiology. We present a method to comprehensively assess the function of single human pluripotent stem cell-derived cardiomyocyte (hPSC-CMs) through simultaneous quantitative analysis of contraction kinetics, force generation, and electrical activity. We demonstrate that statistical analysis of movies of contracting hPSC-CMs can be used to quantify changes in cellular morphology over time and compute contractile kinetics. Using a biomechanical model that incorporates substrate stiffness, we calculate cardiomyocyte force generation at single-cell resolution and validate this approach with conventional traction force microscopy. The addition of fluorescent calcium indicators or membrane potential dyes allows the simultaneous analysis of contractility and calcium handling or action potential morphology. Accordingly, our approach has the potential for broad application in the study of cardiac disease, drug discovery, and cardiotoxicity screening.

  6. Integrated Analysis of Contractile Kinetics, Force Generation, and Electrical Activity in Single Human Stem Cell-Derived Cardiomyocytes

    PubMed Central

    Kijlstra, Jan David; Hu, Dongjian; Mittal, Nikhil; Kausel, Eduardo; van der Meer, Peter; Garakani, Arman; Domian, Ibrahim J.

    2015-01-01

    Summary The quantitative analysis of cardiomyocyte function is essential for stem cell-based approaches for the in vitro study of human cardiac physiology and pathophysiology. We present a method to comprehensively assess the function of single human pluripotent stem cell-derived cardiomyocyte (hPSC-CMs) through simultaneous quantitative analysis of contraction kinetics, force generation, and electrical activity. We demonstrate that statistical analysis of movies of contracting hPSC-CMs can be used to quantify changes in cellular morphology over time and compute contractile kinetics. Using a biomechanical model that incorporates substrate stiffness, we calculate cardiomyocyte force generation at single-cell resolution and validate this approach with conventional traction force microscopy. The addition of fluorescent calcium indicators or membrane potential dyes allows the simultaneous analysis of contractility and calcium handling or action potential morphology. Accordingly, our approach has the potential for broad application in the study of cardiac disease, drug discovery, and cardiotoxicity screening. PMID:26626178

  7. Exploring the Anti-Cancer Activity of Novel Thiosemicarbazones Generated through the Combination of Retro-Fragments: Dissection of Critical Structure-Activity Relationships

    PubMed Central

    Rasko, Nathalie; Potůčková, Eliška; Mrozek-Wilczkiewicz, Anna; Musiol, Robert; Małecki, Jan G.; Sajewicz, Mieczysław; Ratuszna, Alicja; Muchowicz, Angelika; Gołąb, Jakub; Šimůnek, Tomáš; Richardson, Des R.; Polanski, Jaroslaw

    2014-01-01

    Thiosemicarbazones (TSCs) are an interesting class of ligands that show a diverse range of biological activity, including anti-fungal, anti-viral and anti-cancer effects. Our previous studies have demonstrated the potent in vivo anti-tumor activity of novel TSCs and their ability to overcome resistance to clinically used chemotherapeutics. In the current study, 35 novel TSCs of 6 different classes were designed using a combination of retro-fragments that appear in other TSCs. Additionally, di-substitution at the terminal N4 atom, which was previously identified to be critical for potent anti-cancer activity, was preserved through the incorporation of an N4-based piperazine or morpholine ring. The anti-proliferative activity of the novel TSCs were examined in a variety of cancer and normal cell-types. In particular, compounds 1d and 3c demonstrated the greatest promise as anti-cancer agents with potent and selective anti-proliferative activity. Structure-activity relationship studies revealed that the chelators that utilized “soft” donor atoms, such as nitrogen and sulfur, resulted in potent anti-cancer activity. Indeed, the N,N,S donor atom set was crucial for the formation of redox active iron complexes that were able to mediate the oxidation of ascorbate. This further highlights the important role of reactive oxygen species generation in mediating potent anti-cancer activity. Significantly, this study identified the potent and selective anti-cancer activity of 1d and 3c that warrants further examination. PMID:25329549

  8. Ventrolateral Origin of Each Cycle of Rhythmic Activity Generated by the Spinal Cord of the Chick Embryo

    PubMed Central

    Arai, Yoshiyasu; Mentis, George Z.; Wu, Jiang-young; O'Donovan, Michael J.

    2007-01-01

    Background The mechanisms responsible for generating rhythmic motor activity in the developing spinal cord of the chick embryo are poorly understood. Here we investigate whether the activity of motoneurons occurs before other neuronal populations at the beginning of each cycle of rhythmic discharge. Methodology/Principal Findings The spatiotemporal organization of neural activity in transverse slices of the lumbosacral cord of the chick embryo (E8-E11) was investigated using intrinsic and voltage-sensitive dye (VSD) imaging. VSD signals accompanying episodes of activity comprised a rhythmic decrease in light transmission that corresponded to each cycle of electrical activity recorded from the ipsilateral ventral root. The rhythmic signals were widely synchronized across the cord face, and the largest signal amplitude was in the ventrolateral region where motoneurons are located. In unstained slices we recorded two classes of intrinsic signal. In the first, an episode of rhythmic activity was accompanied by a slow decrease in light transmission that peaked in the dorsal horn and decayed dorsoventrally. Superimposed on this signal was a much smaller rhythmic increase in transmission that was coincident with each cycle of discharge and whose amplitude and spatial distribution was similar to that of the VSD signals. At the onset of a spontaneously occurring episode and each subsequent cycle, both the intrinsic and VSD signals originated within the lateral motor column and spread medially and then dorsally. By contrast, following a dorsal root stimulus, the optical signals originated within the dorsal horn and traveled ventrally to reach the lateral motor column. Conclusions/Significance These findings suggest that motoneuron activity contributes to the initiation of each cycle of rhythmic activity, and that motoneuron and/or R-interneuron synapses are a plausible site for the activity-dependent synaptic depression that modeling studies have identified as a critical

  9. Physical mechanism of the vertical electric field generation over active tectonic faults

    NASA Astrophysics Data System (ADS)

    Pulinets, S. A.

    2009-09-01

    The concept of the Global Electric Circuit (GEC) provides an explanation of the existence of a vertical atmospheric electric field and coupling between the ground and ionosphere. Presently, ionospheric physics pays more attention to electric fields and coupling processes in the polar and auroral regions, whereas in other areas the potential difference between the ground and ionosphere usually is not taken into account. Regional processes exist, however, that are able to significantly affect the GEC parameters and through modification of the ionospheric potential to create plasma density irregularities of different scales within the ionosphere. One such source of ionosphere modification is air ionization in the vicinity of active tectonic faults, which takes place due to increased radon emanation. This paper considers the process of local modification of the GEC and corresponding ionospheric variability due to tectonic activity.

  10. Preliminary results of a randomized study (NPC-9902 Trial) on therapeutic gain by concurrent chemotherapy and/or accelerated fractionation for locally advanced nasopharyngeal carcinoma

    SciTech Connect

    Lee, Anne W.M. . E-mail: awmlee@ha.org.hk; Tung, Stewart Y.; Chan, Anthony T.C.; Chappell, Rick; Fu, Y.-T.; Lu, Tai-Xiang; Tan, Terence; Chua, Daniel T.T.; O'Sullivan, Brian; Xu, Shirley L.; Pang, Ellie S.Y.; Sze, W.-M.; Leung, T.-W.; Kwan, W.-H.; Chan, Paddy; Liu, X.-F.; Tan, E.-H.; Sham, Jonathan; Siu, Lillian; Lau, W.-H.

    2006-09-01

    Purpose: To compare the benefit achieved by concurrent chemoradiotherapy (CRT) and/or accelerated fractionation (AF) vs. radiotherapy (RT) alone with conventional fractionation (CF) for patients with T3-4N0-1M0 nasopharyngeal carcinoma (NPC). Methods and Materials: All patients were irradiated with the same RT technique to {>=}66 Gy at 2 Gy per fraction, conventional five fractions/week in the CF and CF+C (chemotherapy) arms, and accelerated six fractions/week in the AF and AF+C arms. The CF+C and AF+C patients were given the Intergroup 0099 regimen (concurrent cisplatin plus adjuvant cisplatin and 5-fluorouracil). Results: Between 1999 and April 2004, 189 patients were randomly assigned; the trial was terminated early because of slow accrual. The median follow-up was 2.9 years. When compared with the CF arm, significant improvement in failure-free survival (FFS) was achieved by the AF+C arm (94% vs. 70% at 3 years, p = 0.008), but both the AF arm and the CF+C arm were insignificant (p {>=} 0.38). Multivariate analyses showed that CRT was a significant factor: hazard ratio (HR) = 0.52 (0.28-0.97), AF per se was insignificant: HR = 0.68 (0.37-1.25); the interaction of CRT by AF was strongly significant (p = 0.006). Both CRT arms had significant increase in acute toxicities (p < 0.005), and the AF+C arm also incurred borderline increase in late toxicities (34% vs. 14% at 3 years, p = 0.05). Conclusions: Preliminary results suggest that concurrent chemoradiotherapy with accelerated fractionation could significantly improve tumor control when compared with conventional RT alone; further confirmation of therapeutic ratio is warranted.

  11. Regulation of pyruvate dehydrogenase activity and citric acid cycle intermediates during high cardiac power generation.

    PubMed

    Sharma, Naveen; Okere, Isidore C; Brunengraber, Daniel Z; McElfresh, Tracy A; King, Kristen L; Sterk, Joseph P; Huang, Hazel; Chandler, Margaret P; Stanley, William C

    2005-01-15

    A high rate of cardiac work increases citric acid cycle (CAC) turnover and flux through pyruvate dehydrogenase (PDH); however, the mechanisms for these effects are poorly understood. We tested the hypotheses that an increase in cardiac energy expenditure: (1) activates PDH and reduces the product/substrate ratios ([NADH]/[NAD(+)] and [acetyl-CoA]/[CoA-SH]); and (2) increases the content of CAC intermediates. Measurements were made in anaesthetized pigs under control conditions and during 15 min of a high cardiac workload induced by dobutamine (Dob). A third group was made hyperglycaemic (14 mm) to stimulate flux through PDH during the high work state (Dob + Glu). Glucose and fatty acid oxidation were measured with (14)C-glucose and (3)H-oleate. Compared with control, the high workload groups had a similar increase in myocardial oxygen consumption ( and cardiac power. Dob increased PDH activity and glucose oxidation above control, but did not reduce the [NADH]/[NAD(+)] and [acetyl-CoA]/[CoA-SH] ratios, and there were no differences between the Dob and Dob + Glu groups. An additional group was treated with Dob + Glu and oxfenicine (Oxf) to inhibit fatty acid oxidation: this increased [CoA-SH] and glucose oxidation compared with Dob; however, there was no further activation of PDH or decrease in the [NADH]/[NAD(+)] ratio. Content of the 4-carbon CAC intermediates succinate, fumarate and malate increased 3-fold with Dob, but there was no change in citrate content, and the Dob + Glu and Dob + Glu + Oxf groups were not different from Dob. In conclusion, compared with normal conditions, at high myocardial energy expenditure (1) the increase in flux through PDH is regulated by activation of the enzyme complex and continues to be partially controlled through inhibition by fatty acid oxidation, and (2) there is expansion of the CAC pool size at the level of 4-carbon intermediates that is largely independent of myocardial fatty acid oxidation.

  12. Anti-malarial activity of a non-piperidine library of next-generation quinoline methanols

    PubMed Central

    2010-01-01

    Background The clinical utility for mefloquine has been eroded due to its association with adverse neurological effects. Better-tolerated alternatives are required. The objective of the present study was the identification of lead compounds that are as effective as mefloquine, but exhibit physiochemical properties likely to render them less susceptible to passage across the blood-brain barrier. Methods A library of drug-like non-piperidine analogs of mefloquine was synthesized. These compounds are diverse in structure and physiochemical properties. They were screened in appropriate in vitro assays and evaluated in terms of their potential as lead compounds. The correlation of specific structural attributes and physiochemical properties with activity was assessed. Results The most potent analogs were low molecular weight unconjugated secondary amines with no heteroatoms in their side-chains. However, these compounds were more metabolically labile and permeable than mefloquine. In terms of physiochemical properties, lower polar surface area, lower molecular weight, more freely rotatable bonds and fewer H-bond acceptors were associated with greater potency. There was no such relationship between activity and LogP, LogD or the number of hydrogen bond donors (HBDs). The addition of an H-bond donor to the side-chain yielded a series of active diamines, which were as metabolically stable as mefloquine but showed reduced permeability. Conclusions A drug-like library of non-piperidine analogs of mefloquine was synthesized. From amongst this library an active lead series of less permeable, but metabolically stable, diamines was identified. PMID:20149249

  13. Therapeutic intraspinal stimulation to generate activity and promote long-term recovery

    PubMed Central

    Mondello, Sarah E.; Kasten, Michael R.; Horner, Philip J.; Moritz, Chet T.

    2014-01-01

    Neuroprosthetic approaches have tremendous potential for the treatment of injuries to the brain and spinal cord by inducing appropriate neural activity in otherwise disordered circuits. Substantial work has demonstrated that stimulation applied to both the central and peripheral nervous system leads to immediate and in some cases sustained benefits after injury. Here we focus on cervical intraspinal microstimulation (ISMS) as a promising method of activating the spinal cord distal to an injury site, either to directly produce movements or more intriguingly to improve subsequent volitional control of the paretic extremities. Incomplete injuries to the spinal cord are the most commonly observed in human patients, and these injuries spare neural tissue bypassing the lesion that could be influenced by neural devices to promote recovery of function. In fact, recent results have demonstrated that therapeutic ISMS leads to modest but sustained improvements in forelimb function after an incomplete spinal cord injury (SCI). This therapeutic spinal stimulation may promote long-term recovery of function by providing the necessary electrical activity needed for neuron survival, axon growth, and synaptic stability. PMID:24578680

  14. Suppression of FOXM1 Transcriptional Activities via a Single-Stranded DNA Aptamer Generated by SELEX.

    PubMed

    Xiang, Qin; Tan, Guixiang; Jiang, Xia; Wu, Kuangpei; Tan, Weihong; Tan, Yongjun

    2017-03-30

    The transcription factor FOXM1 binds to its consensus sequence at promoters through its DNA binding domain (DBD) and activates proliferation-associated genes. The aberrant overexpression of FOXM1 correlates with tumorigenesis and progression of many cancers. Inhibiting FOXM1 transcriptional activities is proposed as a potential therapeutic strategy for cancer treatment. In this study, we obtained a FOXM1-specific single stranded DNA aptamer (FOXM1 Apt) by SELEX with a recombinant FOXM1 DBD protein as the target of selection. The binding of FOXM1 Apt to FOXM1 proteins were confirmed with electrophoretic mobility shift assays (EMSAs) and fluorescence polarization (FP) assays. Phosphorthioate-modified FOXM1 Apt (M-FOXM1 Apt) bound to FOXM1 as wild type FOXM1 Apt, and co-localized with FOXM1 in nucleus. M-FOXM1-Apt abolished the binding of FOXM1 on its consensus binding sites and suppressed FOXM1 transcriptional activities. Compared with the RNA interference of FOXM1 in cancer cells, M-FOXM1 Apt repressed cell proliferation and the expression of FOXM1 target genes without changing FOXM1 levels. Our results suggest that the obtained FOXM1 Apt could be used as a probe for FOXM1 detection and an inhibitor of FOXM1 transcriptional functions in cancer cells at the same time, providing a potential reagent for cancer diagnosis and treatment in the future.

  15. Immune activation generates corticosterone-mediated terminal reproductive investment in a wild bird

    PubMed Central

    Bowers, E. Keith; Bowden, Rachel M.; Sakaluk, Scott K.; Thompson, Charles F.

    2015-01-01

    Despite classical expectations of a trade-off between immune activity and reproduction, an emergent view suggests that individuals experiencing activation of their immune system actually increase reproductive effort and allocation to offspring as a form of terminal investment in response to reduced survival probability. However, the components and mechanisms of increased parental investment following immunostimulation are currently unknown. We hypothesize that increased glucocorticoid production following immunostimulation modulates the increase in reproductive effort that constitutes terminal investment. We activated the immune system of breeding female house wrens (Troglodytes aedon) with an immunogen and cross-fostered the eggs they subsequently produced to separate pre- and post-natal components of maternal investment. Cross-fostering revealed an increase in both pre- and post-natal allocation from immunostimulated females, which was confirmed by quantification of egg constituents and maternal provisioning behavior. The increase in maternal provisioning was mediated, at least in part, by increased corticosterone in these females. Offspring immune responsiveness was also enhanced through transgenerational immune priming via the egg. Thus, our results indicate that maternal immunostimulation induces transgenerational effects on offspring through both pre- and post-natal parental effects, and support an important role for corticosterone in mediating parental investment. PMID:25996862

  16. The potential of a dielectrophoresis activated cell sorter (DACS) as a next generation cell sorter

    NASA Astrophysics Data System (ADS)

    Lee, Dongkyu; Hwang, Bohyun; Kim, Byungkyu

    2016-12-01

    Originally introduced by H. A. Pohl in 1951, dielectrophoretic (DEP) force has been used as a striking tool for biological particle manipulation (or separation) for the last few decades. In particular, dielectrophoresis activated cell sorters (DACSes) have been developed for applications in various biomedical fields. These applications include cell replacement therapy, drug screening and medical diagnostics. Since a DACS does not require a specific bio-marker, it is able to function as a biological particle sorting tool with numerous configurations for various cells [e.g. red blood cells (RBCs), white blood cells (WBCs), circulating tumor cells, leukemia cells, breast cancer cells, bacterial cells, yeast cells and sperm cells]. This article explores current DACS capabilities worldwide, and it also looks at recent developments intended to overcome particular limitations. First, the basic theories are reviewed. Then, representative DACSes based on DEP trapping, traveling wave DEP systems, DEP field-flow fractionation and DEP barriers are introduced, and the strong and weak points of each DACS are discussed. Finally, for the purposes of commercialization, prerequisites regarding throughput, efficiency and recovery rates are discussed in detail through comparisons with commercial cell sorters (e.g. fluorescent activated and magnetic activated cell sorters).

  17. Suppression of FOXM1 Transcriptional Activities via a Single-Stranded DNA Aptamer Generated by SELEX

    PubMed Central

    Xiang, Qin; Tan, Guixiang; Jiang, Xia; Wu, Kuangpei; Tan, Weihong; Tan, Yongjun

    2017-01-01

    The transcription factor FOXM1 binds to its consensus sequence at promoters through its DNA binding domain (DBD) and activates proliferation-associated genes. The aberrant overexpression of FOXM1 correlates with tumorigenesis and progression of many cancers. Inhibiting FOXM1 transcriptional activities is proposed as a potential therapeutic strategy for cancer treatment. In this study, we obtained a FOXM1-specific single stranded DNA aptamer (FOXM1 Apt) by SELEX with a recombinant FOXM1 DBD protein as the target of selection. The binding of FOXM1 Apt to FOXM1 proteins were confirmed with electrophoretic mobility shift assays (EMSAs) and fluorescence polarization (FP) assays. Phosphorthioate-modified FOXM1 Apt (M-FOXM1 Apt) bound to FOXM1 as wild type FOXM1 Apt, and co-localized with FOXM1 in nucleus. M-FOXM1-Apt abolished the binding of FOXM1 on its consensus binding sites and suppressed FOXM1 transcriptional activities. Compared with the RNA interference of FOXM1 in cancer cells, M-FOXM1 Apt repressed cell proliferation and the expression of FOXM1 target genes without changing FOXM1 levels. Our results suggest that the obtained FOXM1 Apt could be used as a probe for FOXM1 detection and an inhibitor of FOXM1 transcriptional functions in cancer cells at the same time, providing a potential reagent for cancer diagnosis and treatment in the future. PMID:28358012

  18. Role for regulated phosphatase activity in generating mitotic oscillations in Xenopus cell-free extracts.

    PubMed

    Zhang, Tongli; Tyson, John J; Novák, Béla

    2013-12-17

    Although current textbook explanations of cell-cycle control in eukaryotes emphasize the periodic activation of cyclin-dependent protein kinases (CDKs), recent experimental observations suggest a significant role for the periodic activation and inactivation of a CDK-counteracting protein phosphatase 2A with a B55δ subunit (PP2A:B55δ), during mitotic cycles in frog-egg extracts and early embryos. In this paper, we extend an earlier mathematical model of embryonic cell cycles to include experimentally motivated roles for PP2A:B55δ and its regulation by Greatwall kinase. Our model is consistent with what is already known about the regulation of CDK and PP2A:B55δ in frog eggs, and it suggests a previously undescribed role for the Greatwall-PP2A:B55δ interaction in creating a toggle switch for activation of the anaphase-promoting complex as embryonic cells exit mitosis and return to interphase.

  19. Enhanced electricity generation by using algae biomass and activated sludge in microbial fuel cell.

    PubMed

    Rashid, Naim; Cui, Yu-Feng; Saif Ur Rehman, Muhammad; Han, Jong-In

    2013-07-01

    Recently, interest is growing to explore low-cost and sustainable means of energy production. In this study, we have exploited the potential of sustainable energy production from wastes. Activated sludge and algae biomass are used as substrates in microbial fuel cell (MFC) to produce electricity. Activated sludge is used at anode as inoculum and nutrient source. Various concentrations (1-5 g/L) of dry algae biomass are tested. Among tested concentrations, 5 g/L (5000 mg COD/L) produced the highest voltage of 0.89 V and power density of 1.78 W/m(2) under 1000 Ω electric resistance. Pre-treated algae biomass and activated sludge are also used at anode. They give low power output than without pre-treatment. Spent algae biomass is tested to replace whole (before oil extraction) algae biomass as a substrate, but it gives low power output. This work has proved the concept of using algae biomass in MFC for high energy output.

  20. A dynamic model for generating actuator specifications for small arms barrel active stabilization

    NASA Astrophysics Data System (ADS)

    Pathak, Anupam; Brei, Diann; Luntz, Jonathan; Lavigna, Chris

    2006-03-01

    Due to stresses encountered in combat, it is known that soldier marksmanship noticeably decreases regardless of prior training. Active stabilization systems in small arms have potential to address this problem to increase soldier survivability and mission effectiveness. The key to success is proper actuator design, but this is highly dependent on proper specification which is challenging due to the human/weapon interaction. This paper presents a generic analytical dynamic model which is capable of defining the necessary actuation specifications for a wide range of small arms platforms. The model is unique because it captures the human interface--shoulder and arm--that introduces the jitter disturbance in addition to the geometry, inertial properties and active stabilization stiffness of the small arms platform. Because no data to date is available for actual shooter-induced disturbance in field conditions, a method is given using the model to back-solve from measured shooting range variability data the disturbance amplitude information relative to the input source (arm or shoulder). As examples of the applicability of the model to various small arms systems, two different weapon systems were investigated: the M24 sniper weapon and the M16 assault rifle. In both cases, model based simulations provided valuable insight into impact on the actuation specifications (force, displacement, phase, frequency) due to the interplay of the human-weapon-active stabilization interface including the effect of shooter-disturbance frequency, disturbance location (shoulder vs. arm), and system parameters (stiffness, barrel rotation).

  1. Plasma Thrombin Generation and Sensitivity to Activated Protein C Among Patients With Myeloma and Monoclonal Gammopathy of Undetermined Significance.

    PubMed

    Crowley, Maeve P; Kevane, Barry; O'Shea, Susan I; Quinn, Shane; Egan, Karl; Gilligan, Oonagh M; Ní Áinle, Fionnuala

    2016-09-01

    The etiology of the prothrombotic state in myeloma has yet to be definitively characterized. Similarly, while recent evidence suggests that patients with monoclonal gammopathy of undetermined significance (MGUS) may also be at increased risk of thrombosis, the magnitude and the etiology of this risk have also yet to be defined. The present study aims to characterize patterns of plasma thrombin generation and sensitivity to the anticoagulant activity of activated protein C (APC) at the time of initial diagnosis of myeloma and in response to therapy in comparison to that observed among patients with MGUS and matched, healthy volunteers. Patients presenting with newly diagnosed/newly relapsed myeloma (n = 8), MGUS (n = 8), and matched healthy volunteers (n = 8) were recruited. Plasma thrombin generation was determined by calibrated automated thrombography. Peak thrombin generation was significantly higher in patients with myeloma (383.4 ± 33.4 nmol/L) and MGUS (353.4 ± 16.5 nmol/L) compared to healthy volunteers (276.7 ± 20.8 nmol/L; P < .05). In the presence of APC, endogenous thrombin potential was significantly lower in control plasma (228.6 ± 44.5 nmol/L × min) than in either myeloma (866.2 ± 241.3 nmol/L × min, P = .01) or MGUS plasma (627 ± 91.5 nmol/L × min, P = .003). Within the myeloma cohort, peak thrombin generation was significantly higher at diagnosis (353.2 ± 15.9 nmol/L) than following completion of the third cycle of therapy (282.1 ± 15.2 nmol/L; P < .005). Moreover, sensitivity to APC increased progressively with each cycle of chemotherapy. Further study of the etiology and evolving patterns of hypercoagulability among patients with these conditions is warranted and may have future implications for thromboprophylaxis strategies.

  2. Enhanced auditory evoked activity to self-generated sounds is mediated by primary and supplementary motor cortices.

    PubMed

    Reznik, Daniel; Ossmy, Ori; Mukamel, Roy

    2015-02-04

    Accumulating evidence demonstrates that responses in auditory cortex to auditory consequences of self-generated actions are modified relative to the responses evoked by identical sounds generated by an external source. Such modifications have been suggested to occur through a corollary discharge sent from the motor system, although the exact neuroanatomical origin is unknown. Furthermore, since tactile input has also been shown to modify responses in auditory cortex, it is not even clear whether the source of such modifications is motor output or somatosensory feedback. We recorded functional magnetic resonance imaging (fMRI) data from healthy human subjects (n = 11) while manipulating the rate at which they performed sound-producing actions with their right hand. In addition, we manipulated the amount of tactile feedback to examine the relative roles of motor and somatosensory cortices in modifying evoked activity in auditory cortex (superior temporal gyrus). We found an enhanced fMRI signal in left auditory cortex during perception of self-generated sounds relative to passive listening to identical sounds. Moreover, the signal difference between active and passive conditions in left auditory cortex covaried with the rate of sound-producing actions and was invariant to the amount of tactile feedback. Together with functional connectivity analysis, our results suggest motor output from supplementary motor area and left primary motor cortex as the source of signal modification in auditory cortex during perception of self-generated sounds. Motor signals from these regions could represent a predictive signal of the expected auditory consequences of the performed action.

  3. Induction of Apoptosis and Antiproliferative Activity of Naringenin in Human Epidermoid Carcinoma Cell through ROS Generation and Cell Cycle Arrest

    PubMed Central

    Jafri, Asif; Ahmad, Sheeba; Afzal, Mohammad; Arshad, Md

    2014-01-01

    A natural predominant flavanone naringenin, especially abundant in citrus fruits, has a wide range of pharmacological activities. The search for antiproliferative agents that reduce skin carcinoma is a task of great importance. The objective of this study was to analyze the anti-proliferative and apoptotic mechanism of naringenin using MTT assay, DNA fragmentation, nuclear condensation, change in mitochondrial membrane potential, cell cycle kinetics and caspase-3 as biomarkers and to investigate the ability to induce reactive oxygen species (ROS) initiating apoptotic cascade in human epidermoid carcinoma A431 cells. Results showed that naringenin exposure significantly reduced the cell viability of A431 cells (p<0.01) with a concomitant increase in nuclear condensation and DNA fragmentation in a dose dependent manner. The intracellular ROS generation assay showed statistically significant (p<0.001) dose-related increment in ROS production for naringenin. It also caused naringenin-mediated epidermoid carcinoma apoptosis by inducing mitochondrial depolarization. Cell cycle study showed that naringenin induced cell cycle arrest in G0/G1 phase of cell cycle and caspase-3 analysis revealed a dose dependent increment in caspase-3 activity which led to cell apoptosis. This study confirms the efficacy of naringenin that lead to cell death in epidermoid carcinoma cells via inducing ROS generation, mitochondrial depolarization, nuclear condensation, DNA fragmentation, cell cycle arrest in G0/G1 phase and caspase-3 activation. PMID:25330158

  4. High Ca2+ load promotes hydrogen peroxide generation via activation of α-glycerophosphate dehydrogenase in brain mitochondria.

    PubMed

    Tretter, Laszlo; Adam-Vizi, Vera

    2012-12-01

    H(2)O(2) generation associated with α-glycerophosphate (α-GP) oxidation was addressed in guinea pig brain mitochondria challenged with high Ca(2+) load (10 μM). Exposure to 10 μM Ca(2+) induced an abrupt 2.5-fold increase in H(2)O(2) release compared to that measured in the presence of a physiological cytosolic Ca(2+) concentration (100 nM) from mitochondria respiring on 5 mM α-GP in the presence of ADP (2 mM). The Ca(2+)-induced stimulation of H(2)O(2) generation was reversible and unaltered by the uniporter blocker Ru 360, indicating that it did not require Ca(2+) uptake into mitochondria. Enhanced H(2)O(2) generation by Ca(2+) was also observed in the absence of ADP when mitochondria exhibited permeability transition pore opening with a decrease in the NAD(P)H level, dissipation of membrane potential, and mitochondrial swelling. Furthermore, mitochondria treated with the pore-forming peptide alamethicin also responded with an elevated H(2)O(2) generation to a challenge with 10 μM Ca(2+). Ca(2+)-induced promotion of H(2)O(2) formation was further enhanced by the complex III inhibitor myxothiazol. With 20 mM α-GP concentration, stimulation of H(2)O(2) formation by Ca(2+) was detected only in the presence, not in the absence, of ADP. It is concluded that α-glycerophosphate dehydrogenase, which is accessible to and could be activated by a rise in the level of cytosolic Ca(2+), makes a major contribution to Ca(2+)-stimulated H(2)O(2) generation. This work highlights a unique high-Ca(2+)-stimulated reactive oxygen species-forming mechanism in association with oxidation of α-GP, which is largely independent of the bioenergetic state and can proceed even in damaged, functionally incompetent mitochondria.

  5. Singular effect of crystallite size on the charge carrier generation and photocatalytic activity of nano-TiO2

    NASA Astrophysics Data System (ADS)

    Strauss, Mathias; Pastorello, Murilo; Sigoli, Fernando A.; Silva, Juliana M. de Souza e.; Mazali, Italo O.

    2014-11-01

    Photocatalytic processes on nanometric titanium oxide have been extensively studied from the standpoint of solar energy utilization and remediation of water and gas streams polluted with organic molecules. It is accepted that TiO2 crystallite size and crystalline phase are among the parameters involved in the control of the photocatalytic activity. However, while changing the catalyst nanoparticle size, other attributes important to assess photocatalytic activity may be modified, making thus difficult to define to which extension the photoactivity changes are related only to size differences. Therefore, aiming at studying exclusively the size effect and the parameters directly related to size on the photocatalytic activity of nanometric TiO2, in this work it was explored a method to synthesize TiO2 nanoparticles with controlled size, highly similar morphology and comparable phase and degree of crystallinity. A set composed of four samples of nano-TiO2 loaded porous Vycor glass, each sample having a specific TiO2 nanoparticle size, was tested on the photoactivated process of depollution of solutions of salicylic acid and methylene blue. The photocatalytic activity observed for the organic compounds tested was inversely proportional to the TiO2 nanoparticle size. An opposite tendency was observed for the generation of OH radicals during photocatalyst illumination, as more radicals are formed on the material containing the larger TiO2 nanoparticles. Results of this study suggest that photocatalytic activity of nano-TiO2 is less favored by the enhanced light absorption response and the higher generation of oxidative species observed for the larger nanoparticles. Better catalysts were obtained when nano-TiO2 exhibited high surface-to-volume ratio and had small recombination volumes, which respectively favors pollutant adsorption-desorption on catalyst surface and reduce the number of recombined charge carriers.

  6. Application of homology modeling to generate CYP1A1 mutants with enhanced activation of the cancer chemotherapeutic prodrug dacarbazine.

    PubMed

    Lewis, Benjamin C; Mackenzie, Peter I; Miners, John O

    2011-11-01

    The chemotherapeutic prodrug dacarbazine (DTIC) has limited efficacy in human malignancies and exhibits numerous adverse effects that arise from systemic exposure to the cytotoxic metabolite. DTIC is activated by CYP1A1 and CYP1A2 catalyzed N-demethylation. However, structural features of these enzymes that confer DTIC N-demethylation have not been characterized. A validated homology model of CYP1A1 was employed to elucidate structure-activity relationships and to engineer CYP1A1 enzymes with altered DTIC activation. In silico docking demonstrated that DTIC orientates proximally to Ser122, Phe123, Asp313, Ala317, Ile386, Tyr259, and Leu496 of human CYP1A1. The site of metabolism is positioned 5.6 Å from the heme iron at an angle of 105.3°. Binding in the active site is stabilized by H-bonding between Tyr259 and the N(2) position of the imidazole ring. Twenty-seven CYP1A1 mutants were generated and expressed in Escherichia coli in yields ranging from 9 to 225 pmol P450/mg. DTIC N-demethylation by the E161K, E256K, and I458V mutants exhibited Michaelis-Menten kinetics, with decreases in K(m) (183-249 μM) that doubled the catalytic efficiency (p < 0.05) relative to wild-type CYP1A1 (K(m), 408 ± 43 μM; V(max), 28 ± 4 pmol · min(-1) · pmol of P450(-1)). The generation of enzymes with catalytically enhanced DTIC activation highlights the potential use of mutant CYP1A1 proteins in P450-based gene-directed enzyme prodrug therapy for the treatment of metastatic malignant melanoma.

  7. Pentosidine in advanced glycation end-products (AGEs) during UVA irradiation generates active oxygen species and impairs human dermal fibroblasts.

    PubMed

    Okano, Y; Masaki, H; Sakurai, H

    2001-08-01

    Our previous study reported that advanced glycation end-products (AGE)-modified BSA produced active oxygen species, *O2-, H2O2, and *OH under UVA irradiation and enhanced the cytotoxicity of UVA light. We examined whether pentosidine in AGE-modified BSA was involved in one of the mechanisms generating the active oxygen species. In biological investigations, fibroblasts exposed to UVA (20 J/cm2) in the presence of pentosidine-rich compounds (PRCs), which were prepared with L-arginine, L-lysine and glucose, showed a time-dependent leakage of the cytosolic enzyme LDH. In addition, release of LDH was suppressed by addition of DMSO and deferoxamine under UVA irradiation. From these results, it was determined that PRCs exposed to UVA damaged the plasma membrane of human dermal fibroblasts due to the conversion of *OH from H2O2 via a Fenton-like reaction. These features of PRCs exposed to UVA were consistent with those of AGE-modified BSA. In an ESR study, PRCs under UVA irradiation yielded DMPO-OH (DMPO-OH adduct) using DMPO as a spin-trapping reagent. *O2- generation from UVA-irradiated PRCs was also indicated by the combination of NBT reduction and SOD. When PRCs were exposed to UVA light controlled with a long-pass filter, WG-360, it was found that their production of *O2- was prohibited less than 50% in the NBT reduction assay. The *O2- production profile of PRCs depending on the wavelength of UVA light was similar to that of AGE-modified BSA. Furthermore, it was found that the H2O2 level was increased by PRCs exposed to UVA. These results indicated that pentosidine is an important factor of AGE-modified BSA in active oxygen generation under UVA irradiation.

  8. ADAMTS metalloproteases generate active versican fragments that regulate interdigital web regression

    PubMed Central

    McCulloch, Daniel R.; Nelson, Courtney M.; Dixon, Laura J.; Silver, Debra L.; Wylie, James D.; Lindner, Volkhard; Sasaki, Takako; Cooley, Marion A.; Argraves, W. Scott; Apte, Suneel S.

    2009-01-01

    We show that combinatorial mouse alleles for the secreted metalloproteases Adamts5, Adamts20 (bt), and Adamts9 result in fully penetrant soft-tissue syndactyly. Interdigital webs in Adamts5−/−; bt/bt mice had reduced apoptosis and decreased cleavage of the proteoglycan versican; however, the BMP-FGF axis, which regulates interdigital apoptosis was unaffected. BMP4 induced apoptosis, but without concomitant versican proteolysis. Haploinsufficiency of either Vcan or Fbln1, a co-factor for versican processing by ADAMTS5, led to highly penetrant syndactyly in bt mice, suggesting that cleaved versican was essential for web regression. The local application of an amino-terminal versican fragment corresponding to ADAMTS-processed versican, induced cell death in Adamts5−/−; bt/bt webs. Thus, ADAMTS proteases cooperatively maintain versican proteolysis above a required threshold to create a permissive environment for apoptosis. The data highlight the developmental significance of proteolytic action on the ECM, not only as a clearance mechanism, but also as a means to generate bioactive versican fragments. PMID:19922873

  9. Trend of photospheric helicity flux in active regions generating halo CMEs

    NASA Astrophysics Data System (ADS)

    Smyrli, Aimilia; Zuccarello, Francesco; Zuccarello, Francesca; Romano, Paolo; Guglielmino, Salvatore Luigi; Spadaro, Daniele; Hood, Alan; Mackay, Duncan

    Coronal Mass Ejections (CMEs) are very energetic events initiated in the solar atmosphere, re-sulting in the expulsion of magnetized plasma clouds that propagate into interplanetary space. It has been proposed that CMEs can play an important role in shedding magnetic helicity, avoiding its endless accumulation in the corona. We therefore investigated the behavior of magnetic helicity accumulation in sites where the initiation of CMEs occurred, in order to de-termine whether and how changes in magnetic helicity accumulation are temporally correlated with CME occurrence. After identifying the active regions (AR) where the CMEs were ini-tiated by means of a double cross-check based on the flaring-eruptive activity and the use of SOHO/EIT difference images, we used MDI magnetograms to calculate magnetic flux evolu-tion, magnetic helicity injection rate and magnetic helicity injection in 10 active regions that gave rise to 12 halo CMEs observed during the period February 2000 -June 2003. No unique behavior in magnetic helicity injection accompanying halo CME occurrence is found. In fact, in some cases there is an abrupt change in helicity injection timely correlated with the CME event, while in some others no significant variation is recorded. However, our analysis show that the most significant changes in magnetic flux and magnetic helicity injection are associated with impulsive CMEs rather than gradual CMEs. Moreover, the most significant changes in mag-netic helicity are observed when X-class flares or eruptive filaments occur, while the occurrence of flares of class C or M seems not to affect significantly the magnetic helicity accumulation.