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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. Model Predictive Controlled Active NPC Inverter for Voltage Stress Balancing Among the Semiconductor Power Switches

    NASA Astrophysics Data System (ADS)

    Parvez Akter, Md.; Dah-Chuan Lu, Dylan

    2017-07-01

    This paper presents a model predictive controlled three-level three-phase active neutral-point-clamped (ANPC) inverter for distributing the voltage stress among the semiconductor power switches as well as balancing the neutral-point voltage. The model predictive control (MPC) concept uses the discrete variables and effectively operates the ANPC inverter by avoiding any linear controller or modulation techniques. A 4.0 kW three-level three-phase ANPC inverter is developed in the MATLAB/Simulink environment to verify the effectiveness of the proposed MPC scheme. The results confirm that the proposed model predictive controlled ANPC inverter equally distributes the voltage across all the semiconductor power switches and provides lowest THD (0.99%) compared with the traditional NPC inverter. Moreover, the neutral-point voltage balancing is accurately maintained by the proposed MPC algorithm. Furthermore, this MPC concept shows the robustness capability against the parameter uncertainties of the system which is also analyzed by MATLAB/Simulink.

  5. Assessment of plasma chitotriosidase activity, CCL18/PARC concentration and NP-C suspicion index in the diagnosis of Niemann-Pick disease type C: a prospective observational study.

    PubMed

    De Castro-Orós, Isabel; Irún, Pilar; Cebolla, Jorge Javier; Rodriguez-Sureda, Victor; Mallén, Miguel; Pueyo, María Jesús; Mozas, Pilar; Dominguez, Carmen; Pocoví, Miguel

    2017-02-21

    Niemann-Pick disease type C (NP-C) is a rare, autosomal recessive neurodegenerative disease caused by mutations in either the NPC1 or NPC2 genes. The diagnosis of NP-C remains challenging due to the non-specific, heterogeneous nature of signs/symptoms. This study assessed the utility of plasma chitotriosidase (ChT) and Chemokine (C-C motif) ligand 18 (CCL18)/pulmonary and activation-regulated chemokine (PARC) in conjunction with the NP-C suspicion index (NP-C SI) for guiding confirmatory laboratory testing in patients with suspected NP-C. In a prospective observational cohort study, incorporating a retrospective determination of NP-C SI scores, two different diagnostic approaches were applied in two separate groups of unrelated patients from 51 Spanish medical centers (n = 118 in both groups). From Jan 2010 to Apr 2012 (Period 1), patients with ≥2 clinical signs/symptoms of NP-C were considered 'suspected NP-C' cases, and NPC1/NPC2 sequencing, plasma chitotriosidase (ChT), CCL18/PARC and sphingomyelinase levels were assessed. Based on findings in Period 1, plasma ChT and CCL18/PARC, and NP-C SI prediction scores were determined in a second group of patients between May 2012 and Apr 2014 (Period 2), and NPC1 and NPC2 were sequenced only in those with elevated ChT and/or elevated CCL18/PARC and/or NP-C SI ≥70. Filipin staining and 7-ketocholesterol (7-KC) measurements were performed in all patients with NP-C gene mutations, where possible. In total across Periods 1 and 2, 10/236 (4%) patients had a confirmed diagnosis o NP-C based on gene sequencing (5/118 [4.2%] in each Period): all of these patients had two causal NPC1 mutations. Single mutant NPC1 alleles were detected in 8/236 (3%) patients, overall. Positive filipin staining results comprised three classical and five variant biochemical phenotypes. No NPC2 mutations were detected. All patients with NPC1 mutations had high ChT activity, high CCL18/PARC concentrations and/or NP-C SI scores ≥70. Plasma 7

  6. 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.

  7. 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

  8. Tafenoquine and NPC-1161B require CYP 2D metabolism for anti-malarial activity: implications for the 8-aminoquinoline class of anti-malarial compounds

    PubMed Central

    2014-01-01

    Background Tafenoquine (TQ) is an 8-aminoquinoline (8AQ) that has been tested in several Phase II and Phase III clinical studies and is currently in late stage development as an anti-malarial prophylactic agent. NPC-1161B is a promising 8AQ in late preclinical development. It has recently been reported that the 8AQ drug primaquine requires metabolic activation by CYP 2D6 for efficacy in humans and in mice, highlighting the importance of pharmacogenomics in the target population when administering primaquine. A logical follow-up study was to determine whether CYP 2D activation is required for other compounds in the 8AQ structural class. Methods In the present study, the anti-malarial activities of NPC-1161B and TQ were assessed against luciferase expressing Plasmodium berghei in CYP 2D knock-out mice in comparison with normal C57BL/6 mice (WT) and with humanized/CYP 2D6 knock-in mice by monitoring luminescence with an in vivo imaging system. These experiments were designed to determine the direct effects of CYP 2D metabolic activation on the anti-malarial efficacy of NPC-1161B and TQ. Results NPC-1161B and TQ exhibited no anti-malarial activity in CYP 2D knock-out mice when dosed at their ED100 values (1 mg/kg and 3 mg/kg, respectively) established in WT mice. TQ anti-malarial activity was partially restored in humanized/CYP 2D6 knock-in mice when tested at two times its ED100. Conclusions The results reported here strongly suggest that metabolism of NPC-1161B and TQ by the CYP 2D enzyme class is essential for their anti-malarial activity. Furthermore, these results may provide a possible explanation for therapeutic failures for patients who do not respond to 8AQ treatment for relapsing malaria. Because CYP 2D6 is highly polymorphic, variable expression of this enzyme in humans represents a significant pharmacogenomic liability for 8AQs which require CYP 2D metabolic activation for efficacy, particularly for large-scale prophylaxis and eradication campaigns. PMID

  9. Tafenoquine and NPC-1161B require CYP 2D metabolism for anti-malarial activity: implications for the 8-aminoquinoline class of anti-malarial compounds.

    PubMed

    Marcsisin, Sean R; Sousa, Jason C; Reichard, Gregory A; Caridha, Diana; Zeng, Qiang; Roncal, Norma; McNulty, Ronan; Careagabarja, Julio; Sciotti, Richard J; Bennett, Jason W; Zottig, Victor E; Deye, Gregory; Li, Qigui; Read, Lisa; Hickman, Mark; Dhammika Nanayakkara, N P; Walker, Larry A; Smith, Bryan; Melendez, Victor; Pybus, Brandon S

    2014-01-03

    Tafenoquine (TQ) is an 8-aminoquinoline (8AQ) that has been tested in several Phase II and Phase III clinical studies and is currently in late stage development as an anti-malarial prophylactic agent. NPC-1161B is a promising 8AQ in late preclinical development. It has recently been reported that the 8AQ drug primaquine requires metabolic activation by CYP 2D6 for efficacy in humans and in mice, highlighting the importance of pharmacogenomics in the target population when administering primaquine. A logical follow-up study was to determine whether CYP 2D activation is required for other compounds in the 8AQ structural class. In the present study, the anti-malarial activities of NPC-1161B and TQ were assessed against luciferase expressing Plasmodium berghei in CYP 2D knock-out mice in comparison with normal C57BL/6 mice (WT) and with humanized/CYP 2D6 knock-in mice by monitoring luminescence with an in vivo imaging system. These experiments were designed to determine the direct effects of CYP 2D metabolic activation on the anti-malarial efficacy of NPC-1161B and TQ. NPC-1161B and TQ exhibited no anti-malarial activity in CYP 2D knock-out mice when dosed at their ED100 values (1 mg/kg and 3 mg/kg, respectively) established in WT mice. TQ anti-malarial activity was partially restored in humanized/CYP 2D6 knock-in mice when tested at two times its ED100. The results reported here strongly suggest that metabolism of NPC-1161B and TQ by the CYP 2D enzyme class is essential for their anti-malarial activity. Furthermore, these results may provide a possible explanation for therapeutic failures for patients who do not respond to 8AQ treatment for relapsing malaria. Because CYP 2D6 is highly polymorphic, variable expression of this enzyme in humans represents a significant pharmacogenomic liability for 8AQs which require CYP 2D metabolic activation for efficacy, particularly for large-scale prophylaxis and eradication campaigns.

  10. Role of EBNA1 in NPC tumourigenesis.

    PubMed

    Frappier, Lori

    2012-04-01

    EBNA1 is expressed in all NPC tumours and is the only Epstein-Barr virus protein needed for the stable persistence of EBV episomes. EBNA1 binds to specific sequences in the EBV genome to facilitate the initiation of DNA synthesis, ensure the even distribution of the viral episomes to daughter cells during mitosis and to activate the transcription of other viral latency genes important for cell immortalization. In addition, EBNA1 has been found to alter cellular pathways in multiple ways that likely contribute to cell immortalization and malignant transformation. This chapter discusses the known functions and cellular effects of EBNA1, especially as pertains to NPC. Copyright © 2011 Elsevier Ltd. All rights reserved.

  11. 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

  12. 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

  13. 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.

  14. 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.

  15. A novel mouse model of Niemann–Pick type C disease carrying a D1005G-Npc1 mutation comparable to commonly observed human mutations

    PubMed Central

    Maue, Robert A.; Burgess, Robert W.; Wang, Bing; Wooley, Christine M.; Seburn, Kevin L.; Vanier, Marie T.; Rogers, Maximillian A.; Chang, Catherine C.; Chang, Ta-Yuan; Harris, Brent T.; Graber, David J.; Penatti, Carlos A.A.; Porter, Donna M.; Szwergold, Benjamin S.; Henderson, Leslie P.; Totenhagen, John W.; Trouard, Theodore P.; Borbon, Ivan A.; Erickson, Robert P.

    2012-01-01

    We have identified a point mutation in Npc1 that creates a novel mouse model (Npc1nmf164) of Niemann–Pick type C1 (NPC) disease: a single nucleotide change (A to G at cDNA bp 3163) that results in an aspartate to glycine change at position 1005 (D1005G). This change is in the cysteine-rich luminal loop of the NPC1 protein and is highly similar to commonly occurring human mutations. Genetic and molecular biological analyses, including sequencing the Npc1spm allele and identifying a truncating mutation, confirm that the mutation in Npc1nmf164 mice is distinct from those in other existing mouse models of NPC disease (Npc1nih, Npc1spm). Analyses of lifespan, body and spleen weight, gait and other motor activities, as well as acoustic startle responses all reveal a more slowly developing phenotype in Npc1nmf164 mutant mice than in mice with the null mutations (Npc1nih, Npc1spm). Although Npc1 mRNA levels appear relatively normal, Npc1nmf164 brain and liver display dramatic reductions in Npc1 protein, as well as abnormal cholesterol metabolism and altered glycolipid expression. Furthermore, histological analyses of liver, spleen, hippocampus, cortex and cerebellum reveal abnormal cholesterol accumulation, glial activation and Purkinje cell loss at a slower rate than in the Npc1nih mouse model. Magnetic resonance imaging studies also reveal significantly less demyelination/dysmyelination than in the null alleles. Thus, although prior mouse models may correspond to the severe infantile onset forms of NPC disease, Npc1nmf164 mice offer many advantages as a model for the late-onset, more slowly progressing forms of NPC disease that comprise the large majority of human cases. PMID:22048958

  16. Dataset in support of the generation of Niemann-Pick disease Type C1 patient-specific iPS cell lines carrying the novel NPC1 mutation c.1180T>C or the prevalent c.3182T>C mutation - Analysis of pluripotency and neuronal differentiation.

    PubMed

    Peter, Franziska; Trilck, Michaela; Rabenstein, Michael; Rolfs, Arndt; Frech, Moritz J

    2017-06-01

    Data presented in this article demonstrate the generation and characterization of two novel Niemann-Pick disease Type C1 (NPC1) patient-specific induced pluripotent stem cell (iPSC) lines, related to the research article Trilck et al. (Diversity of Glycosphingolipid GM2 and Cholesterol Accumulation in NPC1 Patient-Specific iPSC-Derived Neurons; Brain Res.; 2017; 1657:52-61. doi: 10.1016/j.brainres.2016.11.031). For reprogramming fibroblasts, carrying the novel homozygous mutation c.1180T>C and the prevalent homozygous mutation c.3182T>C, were used. Reprogramming into patient-specific iPSCs was induced by retroviral transduction of the transcription factors Sox2, Klf4, Oct4 and c-Myc, and confirmed according to their pluripotency. The iPSCs were subsequently differentiated into neural progenitor cells, which were terminally differentiated into functional neurons and glial cells. The generation of these cell lines provides further valuable tools to investigate pathogenic mechanism of NPC1 in human neuronal cells carrying different NPC1 mutations.

  17. 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.

  18. 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.

  19. 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.

  20. 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

  1. 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.

  2. Human NPC1L1 expression is positively regulated by PPARα.

    PubMed

    Iwayanagi, Yuki; Takada, Tappei; Tomura, Fumiya; Yamanashi, Yoshihide; Terada, Tomohiro; Inui, Ken-ichi; Suzuki, Hiroshi

    2011-02-01

    Niemann-Pick C1-like 1 (NPC1L1), a pharmacological target of ezetimibe, is responsible for cholesterol absorption in enterocytes and hepatocytes. In the present study, the involvement of peroxisome proliferator-activated receptor α (PPARα) and its cofactor, PPARγ coactivator 1α (PGC1α) in the transcriptional regulation of human NPC1L1 was analyzed. Reporter gene assays and electrophoretic mobility shift assays (EMSAs) were performed with the 5'-flanking region of the human NPC1L1 gene and the effect of siPPARα was examined. PPARα-mediated transactivation was observed with human NPC1L1 promoter constructs. Detailed analyses using deletion- and mutated-promoter constructs revealed the presence of a functional PPARα-response element (PPRE) upstream of the human NPC1L1 gene (-846/-834), a direct binding of PPARα and RXRα to which was confirmed by EMSAs. Moreover, PPARα-specific knockdown resulted in a significant decrease in the endogenous expression of NPC1L1 mRNA and protein in human-derived HepG2 cells. Furthermore, cotransfection of PGC1α stimulated the SREBP2/HNF4α- and PPARα/RXRα-mediated activation of the human NPC1L1 promoter. We found that PPARα positively regulates human NPC1L1 transcription via direct binding to a PPRE. Additionally, PGC1α stimulates the SREBP2/HNF4α- and PPARα/RXRα-mediated transactivation of human NPC1L1. These findings may provide new insights into the close relationship of glucose, fatty acids and cholesterol homeostasis.

  3. The phosphatidylcholine-hydrolysing phospholipase C NPC4 plays a role in response of Arabidopsis roots to salt stress.

    PubMed

    Kocourková, Daniela; Krcková, Zuzana; Pejchar, Premysl; Veselková, Stepánka; Valentová, Olga; Wimalasekera, Rinukshi; Scherer, Günther F E; Martinec, Jan

    2011-07-01

    Phosphatidylcholine-hydrolysing phospholipase C, also known as non-specific phospholipase C (NPC), is a new member of the plant phospholipase family that reacts to environmental stresses such as phosphate deficiency and aluminium toxicity, and has a role in root development and brassinolide signalling. Expression of NPC4, one of the six NPC genes in Arabidopsis, was highly induced by NaCl. Maximum expression was observed from 3 h to 6 h after the salt treatment and was dependent on salt concentration. Results of histochemical analysis of P(NPC4):GUS plants showed the localization of salt-induced expression in root tips. On the biochemical level, increased NPC enzyme activity, indicated by accumulation of diacylglycerol, was observed as early as after 30 min of salt treatment of Arabidopsis seedlings. Phenotype analysis of NPC4 knockout plants showed increased sensitivity to salinity as compared with wild-type plants. Under salt stress npc4 plants had shorter roots, lower fresh weight, and reduced seed germination. Expression levels of abscisic acid-related genes ABI1, ABI2, RAB18, PP2CA, and SOT12 were substantially reduced in salt-treated npc4 plants. These observations demonstrate a role for NPC4 in the response of Arabidopsis to salt stress.

  4. 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

  5. Effect of NPC15199 on [Ca²⁺]i and viability in SCM1 human gastric cancer cells.

    PubMed

    Cheng, He-Hsiung; Chou, Chiang-Ting; Liang, Wei-Zhe; Cheng, Jin-Shiung; Chang, Hong-Tai; Kuo, Chun-Chi; Chen, I-S; Lu, Ti; Yu, C-C; Chen, Fu-An; Kuo, Daih-Huang; Shieh, Pochuen; Jan, Chung-Ren

    2016-10-31

    NPC15199 is a synthesized compound that inhibits inflammation in some models. However, whether NPC15199 affects Ca²⁺ homeostasis in human gastric cancer is unclear. This study examined the effect of NPC15199 on cytosolic free Ca²⁺ concentrations ([Ca²⁺]i) and viability in SCM1 human gastric cancer cells. The Ca²⁺-sensitive fluorescent dye fura-2 was used to measure [Ca²⁺]i. NPC15199 evoked [Ca²⁺]i rises concentration-dependently. The response was reduced by removing extracellular Ca²⁺. NPC15199-evoked Ca²⁺ entry was not inhibited by store-operated channel inhibitors (nifedipine, econazole and SKF96365) and protein kinase C (PKC) activator (phorbol 12-myristate 13 acetate, PMA), or PKC inhibitor (GF109203X). In Ca²⁺-free medium, treatment with the endoplasmic reticulum Ca²⁺ pump inhibitor thapsigargin or 2,5-di-tert-butylhydroquinone (BHQ) nearly abolished NPC15199-evoked [Ca²⁺]i rises. Conversely, treatment with NPC15199 also nearly abolished thapsigargin or BHQ-evoked [Ca²⁺]i rises. Inhibition of phospholipase C (PLC) with U73122 did not affect NPC15199-evoked [Ca²⁺]i rises. NPC15199 at concentrations of 100-900 μM induced concentration-dependent, Ca²⁺-independent decrease in viability. Together, in SCM1 cells, NPC15199 induced [Ca²⁺]i rises that involved Ca²⁺ entry through PKC-insensitive non-store-operated Ca²⁺ channels and PLC-independent Ca²⁺ release from the endoplasmic reticulum. NPC15199 also induced Ca²⁺-independent cell death.

  6. 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.

  7. Expression patterns of sterol transporters NPC1 and NPC2 in the cnidarian-dinoflagellate symbiosis.

    PubMed

    Dani, Vincent; Priouzeau, Fabrice; Mertz, Marjolijn; Mondin, Magali; Pagnotta, Sophie; Lacas-Gervais, Sandra; Davy, Simon K; Sabourault, Cécile

    2017-10-01

    The symbiotic interaction between cnidarians (e.g., corals and sea anemones) and photosynthetic dinoflagellates of the genus Symbiodinium is triggered by both host-symbiont recognition processes and metabolic exchange between the 2 partners. The molecular communication is crucial for homeostatic regulation of the symbiosis, both under normal conditions and during stresses that further lead to symbiosis collapse. It is therefore important to identify and fully characterise the key players of this intimate interaction at the symbiotic interface. In this study, we determined the cellular and subcellular localization and expression of the sterol-trafficking Niemann-Pick type C proteins (NPC1 and NPC2) in the symbiotic sea anemones Anemonia viridis and Aiptasia sp. We first established that NPC1 is localised within vesicles in host tissues and to the symbiosome membranes in several anthozoan species. We demonstrated that the canonical NPC2-a protein is mainly expressed in the epidermis, whereas the NPC2-d protein is closely associated with symbiosome membranes. Furthermore, we showed that the expression of the NPC2-d protein is correlated with symbiont presence in healthy symbiotic specimens. As npc2-d is a cnidarian-specific duplicated gene, we hypothesised that it probably arose from a subfunctionalisation process that might result in a gain of function and symbiosis adaptation in anthozoans. Niemann-Pick type C proteins may be key players in a functional symbiosis and be useful tools to study host-symbiont interactions in the anthozoan-dinoflagellate association. © 2017 John Wiley & Sons Ltd.

  8. Neuronal loss of Drosophila NPC1a causes cholesterol aggregation and age-progressive neurodegeneration.

    PubMed

    Phillips, Scott E; Woodruff, E A; Liang, Ping; Patten, Meaghan; Broadie, Kendal

    2008-06-25

    The mistrafficking and consequent cytoplasmic accumulation of cholesterol and sphingolipids is linked to multiple neurodegenerative diseases. One class of disease, the sphingolipid storage diseases, includes Niemann-Pick disease type C (NPC), caused predominantly (95%) by mutation of the NPC1 gene. A disease model has been established through mutation of Drosophila NPC1a (dnpc1a). Null mutants display early lethality attributable to loss of cholesterol-dependent ecdysone steroid hormone production. Null mutants rescued to adults by restoring ecdysone production mimic human NPC patients with progressive motor defects and reduced life spans. Analysis of dnpc1a null brains shows elevated overall cholesterol levels and progressive accumulation of filipin-positive cholesterol aggregates within brain and retina, as well as isolated cultured brain neurons. Ultrastructural imaging of dnpc1a mutant brains reveals age-progressive accumulation of striking multilamellar and multivesicular organelles, preceding the onset of neurodegeneration. Consistently, electroretinogram recordings show age-progressive loss of phototransduction and photoreceptor synaptic transmission. Early lethality, movement impairments, neuronal cholesterol deposits, accumulation of multilamellar bodies, and age-dependent neurodegeneration are all rescued by targeted neuronal expression of a wild-type dnpc1a transgene. Interestingly, targeted expression of dnpc1a in glia also provides limited rescue of adult lethality. Generation of dnpc1a null mutant neuron clones in the brain reveals cell-autonomous requirements for dNPC1a in cholesterol and membrane trafficking. These data demonstrate a requirement for dNPC1a in the maintenance of neuronal function and viability and show that loss of dNPC1a in neurons mimics the human neurodegenerative condition.

  9. Management outcome of NPC-related and non-NPC-related brain abscess in Hong Kong.

    PubMed

    Tam, Mandy Ho-Man; Wong, George Kwok-Chu; Ip, Margaret; Kam, Michael Koon Ming; Abrigo, Jill Morales; Zhu, Xian Lun; Poon, Wai Sang

    2012-07-01

    (1) To review the patient profile, management outcome and prognostic factors of brain abscess; (2) To compare the neurological outcome of nasopharyngeal carcinoma (NPC)-related brain abscess with non-NPC related brain abscess. Retrospective review of consecutive patients diagnosed (radiologically and/or microbiologically) with brain abscess in a regional neurosurgical center in Hong Kong over a nine year period. Fifty-four patients were recruited into this study. There were 37 male and 17 female patients. Eighteen (33%) patients had previous radiotherapy for nasopharyngeal carcinoma. Only 31 (57%) patients had fever on presentation. White cell count and/or C-reactive protein, was raised in 41 (76%) patients on admission. Surgical drainage was carried out in 49 (91%) patients, either by aspiration through a craniotomy, by drainage with corticotomy, or excision of the abscess. Abscess culture was positive in 45 (83%) patients. Common organisms isolated included Streptococcus species (35%) and Peptostreptococcus species (18%). Anaerobes were isolated in 50% of the NPC-related abscesses. The mean follow-up time was 34 months. At the 6 months interval, 24 (44%) patients had good recovery. Favorable outcome was achieved in 30 (55%) patients. NPC-related brain abscess was associated with unfavorable neurological outcome (33%, p = 0.04). There was also a trend towards higher in-patient mortality in patients with NPC-related brain abscess (22%, p = 0.08). Brain abscess carried a substantial morbidity and mortality despite aggressive surgical and medical treatment. Patients with NPC-related brain abscess had a higher mortality and unfavorable neurological outcome. Copyright © 2011 Elsevier B.V. All rights reserved.

  10. 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_

  11. 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.

  12. RNA Export through the NPC in Eukaryotes

    PubMed Central

    Okamura, Masumi; Inose, Haruko; Masuda, Seiji

    2015-01-01

    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. PMID:25802992

  13. 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.

  14. Astrocyte-Only Npc1 Reduces Neuronal Cholesterol and Triples Life Span of Npc1−/− Mice

    PubMed Central

    Zhang, Min; Strnatka, Diana; Donohue, Carolyn; Hallows, Jan; Vincent, Inez; Erickson, Robert P.

    2009-01-01

    Niemann-Pick type C (NPC) disease is an autosomal recessive, lethal neurodegenerative disorder. Although neurodegeneration of Purkinje cells in the mouse model (Npc1−/−) is thought to be autonomous, the basis of neuronal death in other regions of the brain remains elusive. We addressed this issue in vivo by using the glial fibrillary acidic protein (GFAP) promoter to direct astrocyte-specific, replacement expression of Npc1 in Npc1−/− mice. These mice showed enhanced survival, decreased neuronal storage of cholesterol associated with less accumulation of axonal spheroids, lower numbers of degenerated neurons and reactive astrocytes and restoration of myelin tracts. Their death was not associated with the usual terminal decline in weight, but instead with a loss of Purkinje cells and motor coordination. We conclude that neurodegeneration of Npc1−/− mice is greatly affected by the loss of fibrillary astrocyte function. PMID:18500759

  15. 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

  16. 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

  17. 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

  18. 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.

  19. Diversity of glycosphingolipid GM2 and cholesterol accumulation in NPC1 patient-specific iPSC-derived neurons.

    PubMed

    Trilck, Michaela; Peter, Franziska; Zheng, Chaonan; Frank, Marcus; Dobrenis, Kostantin; Mascher, Hermann; Rolfs, Arndt; Frech, Moritz J

    2017-02-15

    Niemann-Pick disease Type C1 (NPC1) is a rare progressive neurodegenerative disorder caused by mutations in the NPC1 gene. On the cellular level NPC1 mutations lead to an accumulation of cholesterol and gangliosides. As a thorough analysis of the severely affected neuronal cells is unfeasible in NPC1 patients, we recently described the cellular phenotype of neuronal cells derived from NPC1 patient iPSCs carrying the compound heterozygous mutation c.1836A>C/c.1628delC. Here we expanded the analysis to cell lines carrying the prevalent mutation c.3182T>C and the novel mutation c.1180T>C, as well as to the determination of GM2 and GM3 gangliosides in NPC1 patient-specific iPSC-derived neurons and glia cells. Immunocytochemical detection of GM2 revealed punctated staining pattern predominantly localized in neurons. Detection of cholesterol by filipin staining showed a comparable staining pattern, colocalized with GM2, indicating a deposit of GM2 and cholesterol in the same cellular compartments. Accumulations were not only restricted to cell bodies, but were also found in the neuronal extensions. A quantification of the GM2 amount by HPLC-MS/MS confirmed significantly higher amounts in neurons carrying a mutation. Additionally, these cells displayed a lowered activity of the catabolic enzyme Hex A, but not B4GALNT1. Molecular docking simulations indicated binding of cholesterol to Hex A, suggesting cholesterol influences the GM2 degradation pathway and, subsequently, leading to the accumulation of GM2. Taken together, this is the first study showing an accumulation of GM2 in neuronal derivatives of patient-specific iPSCs and thus proving further disease-specific hallmarks in this human in vitro model of NPC1. Copyright © 2016 Elsevier B.V. All rights reserved.

  20. 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

  1. Fenofibrate reduces intestinal cholesterol absorption via PPARalpha-dependent modulation of NPC1L1 expression in mouse.

    PubMed

    Valasek, Mark A; Clarke, Stephen L; Repa, Joyce J

    2007-12-01

    Fibrates, including fenofibrate, exert their biological effects by binding peroxisome proliferator-activated receptor alpha (PPARalpha), a member of the nuclear receptor superfamily of ligand-activated transcription factors. Treatment with PPARalpha agonists enhances fatty acid oxidation, decreases plasma triglycerides, and may promote reverse cholesterol transport. In addition, fibrate administration can reduce intestinal cholesterol absorption in patients, although the molecular mechanism for this effect is unknown. Because Niemann-Pick C1-Like 1 (NPC1L1) is already known to be a critical protein for cholesterol absorption, we hypothesized that fenofibrate might modulate NPC1L1 expression to alter intestinal cholesterol transport. Here, we find that fenofibrate-treated wild-type mice have decreased fractional cholesterol absorption (35-47% decrease) and increased fecal neutral sterol excretion (51-83% increase), which correspond to decreased expression of NPC1L1 mRNA and protein (38-66% decrease) in the proximal small intestine. These effects of fenofibrate are dependent on PPARalpha, as Ppar alpha-knockout mice fail to respond like wild-type littermates. Fenofibrate affects the ezetimibe-sensitive pathway and retains the ability to decrease cholesterol absorption and NPC1L1 mRNA expression in chow-fed liver X receptor alpha/beta-double-knockout mice and high-cholesterol- or cholic acid-fed wild-type mice. These data demonstrate that fenofibrate specifically acts via PPARalpha to decrease cholesterol absorption at the level of intestinal NPC1L1 expression.

  2. 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.

  3. 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

  4. 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

  5. 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.

  6. 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.

  7. 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

  8. 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

  9. 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

  10. 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.

  11. 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

  12. 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.

  13. 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.

  14. 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.

  15. 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.

  16. Psychiatric and neurological symptoms in patients with Niemann-Pick disease type C (NP-C): findings from the International NPC Registry.

    PubMed

    Bonnot, Olivier; Gama, Clarissa S; Mengel, Eugen; Pineda, Mercè; Vanier, Marie T; Watson, Louise; Watissée, Marie; Schwierin, Barbara; Patterson, Marc C

    2017-09-15

    Niemann-Pick disease type C (NP-C) is a rare inherited neurovisceral disease that should be recognized by psychiatrists as a possible underlying cause of psychiatric abnormalities. This study describes NP-C patients who had psychiatric manifestations at enrolment in the international NPC Registry, a unique multicentre, prospective, observational disease registry. Treating physicians' data entries describing psychiatric manifestations in NPC patients were coded and grouped by expert psychiatrists. Out of 386 NP-C patients included in the Registry as of October 2015, psychiatric abnormalities were reported to be present in 34% (94/280) of those with available data. Forty-four patients were confirmed to have identifiable psychiatric manifestations, with text describing these psychiatric manifestations. In these 44 patients, the median (range) age at onset of psychiatric manifestations was 17.9 years (2.5-67.9; n = 15), while the median (range) age at NP-C diagnosis was 23.7 years (0.2-69.8; n = 34). Almost all patients (43/44 [98%]) had an occurrence of ≥1 neurological manifestation at enrolment. These data show that substantial delays in diagnosis of NP-C are long among patients with psychiatric symptoms and, moreover, patients presenting with psychiatric features and at least one of cognitive impairment, neurological manifestations, and/or visceral symptoms should be screened for NP-C.

  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. 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.

  19. Bradykinin promotes neuron-generating division of neural progenitor cells through ERK activation.

    PubMed

    Pillat, Micheli M; Lameu, Claudiana; Trujillo, Cleber A; Glaser, Talita; Cappellari, Angélica R; Negraes, Priscilla D; Battastini, Ana M O; Schwindt, Telma T; Muotri, Alysson R; Ulrich, Henning

    2016-09-15

    During brain development, cells proliferate, migrate and differentiate in highly accurate patterns. In this context, published results indicate that bradykinin functions in neural fate determination, favoring neurogenesis and migration. However, mechanisms underlying bradykinin function are yet to be explored. Our findings indicate a previously unidentified role for bradykinin action in inducing neuron-generating division in vitro and in vivo, given that bradykinin lengthened the G1-phase of the neural progenitor cells (NPC) cycle and increased TIS21 (also known as PC3 and BTG2) expression in hippocampus from newborn mice. This role, triggered by activation of the kinin-B2 receptor, was conditioned by ERK1/2 activation. Moreover, immunohistochemistry analysis of hippocampal dentate gyrus showed that the percentage of Ki67(+) cells markedly increased in bradykinin-treated mice, and ERK1/2 inhibition affected this neurogenic response. The progress of neurogenesis depended on sustained ERK phosphorylation and resulted in ERK1/2 translocation to the nucleus in NPCs and PC12 cells, changing expression of genes such as Hes1 and Ngn2 (also known as Neurog2). In agreement with the function of ERK in integrating signaling pathways, effects of bradykinin in stimulating neurogenesis were reversed following removal of protein kinase C (PKC)-mediated sustained phosphorylation. © 2016. Published by The Company of Biologists Ltd.

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

    USDA-ARS?s Scientific Manuscript database

    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...

  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. 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

  7. Hydride generation activity of arsenosugars and thioarsenicals.

    PubMed

    Regmi, Rajendra; Milne, Bruce F; Feldmann, Jörg

    2007-06-01

    The major arsenosugar compounds have been reported to be hydride-generation-active, however to a lesser extent in comparison with the inorganic arsenicals. We report here for the first time the identity and quantity of the volatile arsenicals generated by As-sugar-SO(3), As-sugar-SO(4), dimethylarsinoyl acetic acid and dimethylarsinoyl ethanol. Only one major volatile compound was identified for all four compounds studied: dimethylarsine. This means that the As-C bond to the longer carbon chain was cleaved during the hydride-generation process. Theoretical calculations at the RHF/6-31G(d,p) ab initio level confirm that this As-C bond is much weaker than the As-CH(3) bonds. Furthermore, it was revealed that the sulphur analogue of dimethylarsinic acid (DMAS ) is hydride-generation-active at pH 7 in contrast to dimethylarsinic acid, despite the fact that arsenic is also pentavalent. This has been substantiated by the calculation of the change in susceptibility of the arsenic towards nucleophilic attack when oxygen is replaced by sulphur. Hence, DMAS can easily be mistaken for a trivalent arsenic species.

  8. 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.

  9. Swim Pressure: Stress Generation in Active Matter

    NASA Astrophysics Data System (ADS)

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

    2014-07-01

    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.

  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. 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).

  12. 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

  13. 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.

  14. 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.

  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. 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...

  18. 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

  19. 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.

  20. Associations between HLA Class I alleles and the prevalence of nasopharyngeal carcinoma (NPC) among Tunisians

    PubMed Central

    Li, Xin; Ghandri, Nahla; Piancatelli, Daniela; Adams, Sharon; Chen, Deborah; Robbins, Fu-Meei; Wang, Ena; Monaco, Alessandro; Selleri, Silvia; Bouaouina, Noureddine; Stroncek, David; Adorno, Domenico; Chouchane, Lotfi; Marincola, Francesco M

    2007-01-01

    The high prevalence of nasopharyngeal cancer (NPC) in Southern Asia and Mediterranean Northern Africa suggests genetic predisposition among other factors. While Human Leukocyte Antigen (HLA) haplotypes have been conclusively associated with NPC predisposition in Asians, Northern African Maghrebians have been less intensely studied. However, low resolution serological methods identified weak positive associations with HLA-B5, B13 and B18 and a negative with HLA-B14. Using sequence based typing (SBT), we performed a direct comparison of HLA class I frequencies in a cohort of 136 Tunisian patients with NPC matched for gender, age and geographical residence to 148 normal Tunisians. The bimodal age distribution of NPC in Maghrebians was also taken into account. HLA frequencies in normal Tunisians were also compared with those of Northern Moroccan Berbers (ME) to evaluate whether the Tunisian population in this study could be considered representative of other Maghrebian populations. HLA-B14 and -Cw08 were negatively associated with NPC (odd ratio = 0.09 and 0.18 respectively, Fisher p2-value = 0.0001 and = 0.003). Moreover, positive associations were observed for HLA-B-18, -B51 (split of -B5) and -B57 (p2-value < 0.025 in all) confirming previous findings in Maghrebs. The HLA-B14/Cw*08 haplotype frequency (HF) was 0.007 in NPC patients compared to 0.057 in both Tunisian (OR = 0.12; p2-value = 0.001) and Moroccan controls. This study confirms several previous associations noted by serologic typing between HLA class I alleles and the prevalence of NPC in Maghrebians populations. In addition, we identified a putative haplotype rare in Tunisian patients with NPC that may serve as a genetic marker for further susceptibility studies. PMID:17480220

  1. Subversion of NPC1 pathway of cholesterol transport by Anaplasma phagocytophilum

    PubMed Central

    Xiong, Qingming; Rikihisa, Yasuko

    2013-01-01

    Summary Intracellular cholesterol amounts, distribution, and traffic are tightly regulated to maintain the healthy eukaryotic cell function. However, how intracellular pathogens that require cholesterol, interact with the host cholesterol homeostasis and traffic is not well understood. Anaplasma phagocytophilum is an obligatory intracellular and cholesterol-robbing bacterium, which causes human granulocytic anaplasmosis. Here we found that a subset of cholesterol-binding membrane protein, Niemann–Pick type C1 (NPC1)-bearing vesicles devoid of lysosomal markers were upregulated in HL-60 cells infected with A. phagocytophilum, and trafficked to live bacterial inclusions. The NPC1 localization to A. phagocytophilum inclusions was abolished by low-density lipoprotein (LDL)-derived cholesterol traffic inhibitor U18666A. Studies using NPC1 siRNA and the cell line with cholesterol traffic defect demonstrated that the NPC1 function is required for bacterial cholesterol acquisition and infection. Furthermore, trans-Golgi network-specific soluble N-ethylmaleimide-sensitive factor attachment protein receptors, vesicle-associated membrane protein (VAMP4) and syntaxin 16, which are associated with NPC1 and LDL-derived cholesterol vesicular transport were recruited to A. phagocytophilum inclusions, and VAMP4 was required for bacteria infection. Taken together, A. phagocytophilum is the first example of a pathogen that subverts the NPC1 pathway of intracellular cholesterol transport and homeostasis for bacterial inclusion membrane biogenesis and cholesterol capture. PMID:22212234

  2. Ebola virus and severe acute respiratory syndrome coronavirus display late cell entry kinetics: evidence that transport to NPC1+ endolysosomes is a rate-defining step.

    PubMed

    Mingo, Rebecca M; Simmons, James A; Shoemaker, Charles J; Nelson, Elizabeth A; Schornberg, Kathryn L; D'Souza, Ryan S; Casanova, James E; White, Judith M

    2015-03-01

    Ebola virus (EBOV) causes hemorrhagic fevers with high mortality rates. During cellular entry, the virus is internalized by macropinocytosis and trafficked through endosomes until fusion between the viral and an endosomal membrane is triggered, releasing the RNA genome into the cytoplasm. We found that while macropinocytotic uptake of filamentous EBOV viruslike particles (VLPs) expressing the EBOV glycoprotein (GP) occurs relatively quickly, VLPs only begin to enter the cytoplasm after a 30-min lag, considerably later than particles bearing the influenza hemagglutinin or GP from lymphocytic choriomeningitis virus, which enter through late endosomes (LE). For EBOV, the long lag is not due to the large size or unusual shape of EBOV filaments, the need to prime EBOV GP to the 19-kDa receptor-binding species, or a need for unusually low endosomal pH. In contrast, since we observed that EBOV entry occurs upon arrival in Niemann-Pick C1 (NPC1)-positive endolysosomes (LE/Lys), we propose that trafficking to LE/Lys is a key rate-defining step. Additional experiments revealed, unexpectedly, that severe acute respiratory syndrome (SARS) S-mediated entry also begins only after a 30-min lag. Furthermore, although SARS does not require NPC1 for entry, SARS entry also begins after colocalization with NPC1. Since the only endosomal requirement for SARS entry is cathepsin L activity, we tested and provide evidence that NPC1(+) LE/Lys have higher cathepsin L activity than LE, with no detectable activity in earlier endosomes. Our findings suggest that both EBOV and SARS traffic deep into the endocytic pathway for entry and that they do so to access higher cathepsin activity. Ebola virus is a hemorrhagic fever virus that causes high fatality rates when it spreads from zoonotic vectors into the human population. Infection by severe acute respiratory syndrome coronavirus (SARS-CoV) causes severe respiratory distress in infected patients. A devastating outbreak of EBOV occurred in West

  3. Ebola Virus and Severe Acute Respiratory Syndrome Coronavirus Display Late Cell Entry Kinetics: Evidence that Transport to NPC1+ Endolysosomes Is a Rate-Defining Step

    PubMed Central

    Mingo, Rebecca M.; Simmons, James A.; Shoemaker, Charles J.; Nelson, Elizabeth A.; Schornberg, Kathryn L.; D'Souza, Ryan S.; Casanova, James E.

    2014-01-01

    ABSTRACT Ebola virus (EBOV) causes hemorrhagic fevers with high mortality rates. During cellular entry, the virus is internalized by macropinocytosis and trafficked through endosomes until fusion between the viral and an endosomal membrane is triggered, releasing the RNA genome into the cytoplasm. We found that while macropinocytotic uptake of filamentous EBOV viruslike particles (VLPs) expressing the EBOV glycoprotein (GP) occurs relatively quickly, VLPs only begin to enter the cytoplasm after a 30-min lag, considerably later than particles bearing the influenza hemagglutinin or GP from lymphocytic choriomeningitis virus, which enter through late endosomes (LE). For EBOV, the long lag is not due to the large size or unusual shape of EBOV filaments, the need to prime EBOV GP to the 19-kDa receptor-binding species, or a need for unusually low endosomal pH. In contrast, since we observed that EBOV entry occurs upon arrival in Niemann-Pick C1 (NPC1)-positive endolysosomes (LE/Lys), we propose that trafficking to LE/Lys is a key rate-defining step. Additional experiments revealed, unexpectedly, that severe acute respiratory syndrome (SARS) S-mediated entry also begins only after a 30-min lag. Furthermore, although SARS does not require NPC1 for entry, SARS entry also begins after colocalization with NPC1. Since the only endosomal requirement for SARS entry is cathepsin L activity, we tested and provide evidence that NPC1+ LE/Lys have higher cathepsin L activity than LE, with no detectable activity in earlier endosomes. Our findings suggest that both EBOV and SARS traffic deep into the endocytic pathway for entry and that they do so to access higher cathepsin activity. IMPORTANCE Ebola virus is a hemorrhagic fever virus that causes high fatality rates when it spreads from zoonotic vectors into the human population. Infection by severe acute respiratory syndrome coronavirus (SARS-CoV) causes severe respiratory distress in infected patients. A devastating outbreak of EBOV

  4. 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.

  5. 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

  6. [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.

  7. Role of STARD4 and NPC1 in intracellular sterol transport.

    PubMed

    Maxfield, Frederick R; Iaea, David B; Pipalia, Nina H

    2016-12-01

    Cholesterol plays an important role in determining the biophysical properties of membranes in mammalian cells, and the concentration of cholesterol in membranes is tightly regulated. Cholesterol moves among membrane organelles by a combination of vesicular and nonvesicular transport pathways, but the details of these transport pathways are not well understood. In this review, we discuss the mechanisms for nonvesicular sterol transport with an emphasis on the role of STARD4, a small, soluble, cytoplasmic sterol transport protein. STARD4 can rapidly equilibrate sterol between membranes, especially membranes with anionic lipid headgroups. We also discuss the sterol transport in late endosomes and lysosomes, which is mediated by a soluble protein, NPC2, and a membrane protein, NPC1. Homozygous mutations in these proteins lead to a lysosomal lipid storage disorder, Niemann-Pick disease type C. Many of the disease-causing mutations in NPC1 are associated with degradation of the mutant NPC1 proteins in the endoplasmic reticulum. Several histone deacetylase inhibitors have been found to rescue the premature degradation of the mutant NPC1 proteins, and one of these is now in a small clinical trial.

  8. Luminol chemiluminescence and active oxygen generation by activated neutrophils.

    PubMed

    Takahashi, R; Edashige, K; Sato, E F; Inoue, M; Matsuno, T; Utsumi, K

    1991-03-01

    Upon stimulation by various ligands and membrane perturbers, neutrophils produce various active oxygen species. Since luminol chemiluminescence (LCL) in neutrophils can be blocked by azide, an inhibitor of myeloperoxidase, LCL has been believed to reflect mainly the myeloperoxidase-catalyzed reaction. When cells were stimulated by formyl-methionyl-leucyl-phenylalanine, LCL was strongly inhibited by superoxide dismutase (SOD) and uric acid, a scavenger for hydroxy radical (.OH) and singlet oxygen, whereas it was stimulated by azide. LCL was also inhibited by .OH scavengers, such as mannitol, ethanol, and dimethylsulfoxide. However, when stimulated by phorbol myristate acetate or opsonized zymosan, LCL was strongly inhibited by azide but not by uric acid, and the inhibitory action of SOD was low. Thus, the qualitative and quantitative aspects of reactive oxygen generation by activated neutrophils differ significantly from one ligand to another. These results suggest that the metabolic fate of active oxygens in neutrophils and, hence, their effect on microorganisms and the surrounding tissues might differ depending on the stimulus.

  9. Identification of mutation in NPC2 by exome sequencing results in diagnosis of Niemann-Pick disease type C.

    PubMed

    Alavi, Afagh; Nafissi, Shahriar; Shamshiri, Hosein; Nejad, Maryam Malakooti; Elahi, Elahe

    2013-01-01

    We report identification of a homozygous mutation in NPC2 in two Iranian siblings with a neurologic dysfunction whose disease had not been diagnosed prior to our genetic analysis. The mutation was identified by exome sequencing. The finding resulted in diagnosis of Niemann-Pick disease type C (NPC) in the siblings, and initiation of treatment with Miglustat. The clinical features of the patients are presented. It has been suggested that NPC is under diagnosed, particularly when presentations are not very severe, as was the situation in the cases studied here. NPC is a fatal autosomal recessive disorder clinically characterized by hepatosplenomegaly and progressive neurological deterioration. At the cellular level, it causes aberrant cholesterol trafficking and accumulation of unesterified cholesterol in lysosomes. Mutations in NPC1 and NPC2 are cause of disease in respectively, 95% and 5% of NPC patients. The p.Pro120Ser causing mutation in NPC2 observed in the Iranian patients was earlier observed in the only other NPC2 patient reported from the Middle East. The study demonstrates that in addition to greatly facilitating gene discovery, exome sequencing has notable potentials for diagnosis, particularly for diagnosis of atypical cases. Copyright © 2013 Elsevier Inc. All rights reserved.

  10. 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.

  11. Sequential recruitment of NPC proteins to the nuclear periphery at the end of mitosis.

    PubMed

    Bodoor, K; Shaikh, S; Salina, D; Raharjo, W H; Bastos, R; Lohka, M; Burke, B

    1999-07-01

    Nuclear pore complexes (NPCs) are extremely elaborate structures that mediate the bidirectional movement of macromolecules between the nucleus and cytoplasm. With a mass of about 125 MDa, NPCs are thought to be composed of 50 or more distinct protein subunits, each present in multiple copies. During mitosis in higher cells the nuclear envelope is disassembled and its components, including NPC subunits, are dispersed throughout the mitotic cytoplasm. At the end of mitosis, all of these components are reutilized. Using both conventional and digital confocal immunofluorescence microscopy we have been able to define a time course of post-mitotic assembly for a group of NPC components (CAN/Nup214, Nup153, POM121, p62 and Tpr) relative to the integral nuclear membrane protein LAP2 and the NPC membrane glycoprotein gp210. Nup153, a component of the nuclear basket, associates with chromatin towards the end of anaphase, in parallel with the inner nuclear membrane protein, LAP2. However, immunogold labeling suggests that the initial Nup153 chromatin association is membrane-independent. Assembly of the remaining proteins follows that of the nuclear membranes and occurs in the sequence POM121, p62, CAN/Nup214 and gp210/Tpr. Since p62 remains as a complex with three other NPC proteins (p58, 54, 45) during mitosis and CAN/Nup214 maintains a similar interaction with its partner, Nup84, the relative timing of assembly of these additional four proteins may also be inferred. These observations suggest that there is a sequential association of NPC proteins with chromosomes during nuclear envelope reformation and the recruitment of at least eight of these precedes that of gp210. These findings support a model in which it is POM121 rather than gp210 that defines initial membrane-associated NPC assembly intermediates.

  12. 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.

  13. 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.

  14. 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

  15. 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.

  16. 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

  17. 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.

  18. Structural basis of sterol binding by NPC2, a lysosomal protein deficient in Niemann-Pick type C2 disease.

    PubMed

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

    2007-08-10

    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.

  19. 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.

  20. 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.

  1. Cohort study of neurocognitive functioning and adaptive behavior in children and adolescents with NPC1

    PubMed Central

    THURM, AUDREY; FARMER, CRISTAN; FARHAT, NICOLE YANJANIN; WIGGS, EDYTHE; BLACK, DAVID; PORTER, FORBES D

    2015-01-01

    AIM To describe the neurocognitive and adaptive behavior profile of children and adolescents with Niemann–Pick Disease type C1 (NPC1), a rare genetic disease that frequently presents in childhood, with variable onset and symptom complex involving neurodegeneration. METHOD Thirty-eight participants (20 males, 18 females; mean age 8y 10mo, SD 4y 8mo, range 1–18y) with NPC1 were evaluated through a natural history protocol. RESULTS NPC1 severity was in the mild to moderate range for most participants. Cognitive scores (n=32) ranged from very low to above average; about half of the participants exhibited a clinically significant advantage of Verbal IQ over Non-verbal IQ. Adaptive behavior scores (n=21) were generally in the borderline to impaired range. Longitudinal cognitive data (n=19) suggested a pattern of decreasing scores over time. However, most participants remained at the same general level of functioning throughout the study. INTERPRETATION This study begins to systematically describe the neurocognitive phenotype of children and adolescents with NPC1, identifying heterogeneity and decline, aiding in understanding the natural history of the disease to plan treatment studies. PMID:26586413

  2. Generation and Biological Activities of Oxidized Phospholipids

    PubMed Central

    Oskolkova, Olga V.; Birukov, Konstantin G.; Levonen, Anna-Liisa; Binder, Christoph J.; Stöckl, Johannes

    2010-01-01

    Abstract Glycerophospholipids represent a common class of lipids critically important for integrity of cellular membranes. Oxidation of esterified unsaturated fatty acids dramatically changes biological activities of phospholipids. Apart from impairment of their structural function, oxidation makes oxidized phospholipids (OxPLs) markers of “modified-self” type that are recognized by soluble and cell-associated receptors of innate immunity, including scavenger receptors, natural (germ line-encoded) antibodies, and C-reactive protein, thus directing removal of senescent and apoptotic cells or oxidized lipoproteins. In addition, OxPLs acquire novel biological activities not characteristic of their unoxidized precursors, including the ability to regulate innate and adaptive immune responses. Effects of OxPLs described in vitro and in vivo suggest their potential relevance in different pathologies, including atherosclerosis, acute inflammation, lung injury, and many other conditions. This review summarizes current knowledge on the mechanisms of formation, structures, and biological activities of OxPLs. Furthermore, potential applications of OxPLs as disease biomarkers, as well as experimental therapies targeting OxPLs, are described, providing a broad overview of an emerging class of lipid mediators. Antioxid. Redox Signal. 12, 1009–1059. PMID:19686040

  3. 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.

  4. RNA sequencing of esophageal adenocarcinomas identifies novel fusion transcripts, including NPC1-MELK, arising from a complex chromosomal rearrangement.

    PubMed

    Wang, Zhixiong; Cheng, Yulan; Abraham, John M; Yan, Rong; Liu, Xi; Chen, Wei; Ibrahim, Sariat; Schroth, Gary P; Ke, Xiquan; He, Yulong; Meltzer, Stephen J

    2017-06-22

    Studies of chromosomal rearrangements and fusion transcripts have elucidated mechanisms of tumorigenesis and led to targeted cancer therapies. This study was aimed at identifying novel fusion transcripts in esophageal adenocarcinoma (EAC). To identify new fusion transcripts associated with EAC, targeted RNA sequencing and polymerase chain reaction (PCR) verification were performed in 40 EACs and matched nonmalignant specimens from the same patients. Genomic PCR and Sanger sequencing were performed to find the breakpoint of fusion genes. Five novel in-frame fusion transcripts were identified and verified in 40 EACs and in a validation cohort of 15 additional EACs (55 patients in all): fibroblast growth factor receptor 2 (FGFR2)-GRB2-associated binding protein 2 (GAB2) in 2 of 55 or 3.6%, Niemann-Pick C1 (NPC1)-maternal embryonic leucine zipper kinase (MELK) in 2 of 55 or 3.6%, ubiquitin-specific peptidase 54 (USP54)-calcium/calmodulin dependent protein kinase II γ (CAMK2G) in 2 of 55 or 3.6%, megakaryoblastic leukemia (translocation) 1 (MKL1)-fibulin 1 (FBLN1) in 1 of 55 or 1.8%, and CCR4-NOT transcription complex subunit 2 (CNOT2)-chromosome 12 open reading frame 49 (C12orf49) in 1 of 55 or 1.8%. A genomic analysis indicated that NPC1-MELK arose from a complex interchromosomal translocation event involving chromosomes 18, 3, and 9 with 3 rearrangement points, and this was consistent with chromoplexy. These data indicate that fusion transcripts occur at a stable frequency in EAC. Furthermore, our results indicate that chromoplexy is an underlying mechanism that generates fusion transcripts in EAC. These and other fusion transcripts merit further study as diagnostic markers and potential therapeutic targets in EAC. Cancer 2017. © 2017 American Cancer Society. © 2017 American Cancer Society.

  5. 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.

  6. 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

  7. The implementation in-house dose verification for IMRT and VMAT on breast cancer and NPC cases

    NASA Astrophysics Data System (ADS)

    Azzi, A.; Dimitri, F. Y.; Pawiro, S. A.

    2016-03-01

    IMRT and VMAT have been widely implemented on breast cancer and NPC cases. Dose verification was conducted to ensure the comparison between dose delivered and prescribe dose in TPS to obtain optimal results and the dose delivered. The gafchromic films measurement was done for breast and NPC simulation on RANDO Phantom, irradiating film by 6 MV photon with 200 cGy and scanned with EPSON V700 using 72 dpi. Dose distribution was calculated using the MATLAB-based in-house algorithm. In order to evaluate the gamma index, gafchromic films and MatriXX 2D array were placed in the Multi Cube and irradiated with EPID in same position. Dose distribution differences for IMRT and VMAT with modalities to TPS on film was 5.34% and 6.68% for NPC PTV70; 12.81% and 6.15% for NPC PTV50; 11.14% and 10.80% for breast cancer. Gamma index differences on IMRT with modalities to MatriXX 2D array and EPID was -0.09% and 5.13% for NPC; 0.51% and 2.28% for breast cancer. On the other hand, gamma index difference on VMAT was -1.65% and 1.43% for NPC; 0.19% and 4.38% for breast cancer. This measurement method can be used to verify dose for breast cancer and NPC cases in IMRT and VMAT.

  8. 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.

  9. 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.

  10. Activity cliffs and activity cliff generators based on chemotype-related activity landscapes.

    PubMed

    Pérez-Villanueva, Jaime; Méndez-Lucio, Oscar; Soria-Arteche, Olivia; Medina-Franco, José L

    2015-11-01

    Activity cliffs have large impact in drug discovery; therefore, their detection and quantification are of major importance. This work introduces the metric activity cliff enrichment factor and expands the previously reported activity cliff generator concept by adding chemotype information to representations of the activity landscape. To exemplify these concepts, three molecular databases with multiple biological activities were characterized. Compounds in each database were grouped into chemotype classes. Then, pairwise comparisons of structure similarities and activity differences were calculated for each compound and used to construct chemotype-based structure-activity similarity (SAS) maps. Different landscape distributions among four major regions of the SAS maps were observed for different subsets of molecules grouped in chemotypes. Based on this observation, the activity cliff enrichment factor was calculated to numerically detect chemotypes enriched in activity cliffs. Several chemotype classes were detected having major proportion of activity cliffs than the entire database. In addition, some chemotype classes comprising compounds with smooth structure activity relationships (SAR) were detected. Finally, the activity cliff generator concept was applied to compounds grouped in chemotypes to extract valuable SAR information.

  11. Utilizing the Dyn2 dimerization-zipper as a tool to probe NPC structure and function.

    PubMed

    Flemming, Dirk; Stelter, Philipp; Hurt, Ed

    2014-01-01

    The discovery of dynein light chain 2 (Dyn2) as a member of the nucleoporins in yeast led to a series of applications to study NPC structure and function. Its intriguing ability to act as a hub for the parallel dimerization of two short amino acid sequence motifs (DID) prompted us to utilize it as a tool for probing nucleocytoplasmic transport in vivo. Further, the distinct structure of the Dyn2-DID rod, which is easily visible in the electron microscope, allowed us to develop a precise structural label on proteins or protein complexes. This label was used to identify the position of subunits in NPC subcomplexes or to derive at pseudo-atomic models of single large Nups. The versatility for various applications of the DID-Dyn2 system makes it an attractive molecular tool beyond the nuclear pore and transport field. Copyright © 2014 Elsevier Inc. All rights reserved.

  12. 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. Copyright © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  13. 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

  14. 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…

  15. 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…

  16. Niemann-Pick C1-Like 1 (NPC1L1) Protein in Intestinal and Hepatic Cholesterol Transport

    PubMed Central

    Jia, Lin; Betters, Jenna L.; Yu, Liqing

    2014-01-01

    Increased blood cholesterol is an independent risk factor for atherosclerotic cardiovascular disease. Cholesterol homeostasis in the body is controlled mainly by endogenous synthesis, intestinal absorption, and hepatic excretion. Niemann-Pick C1-Like 1 (NPC1L1) is a polytopic transmembrane protein localized at the apical membrane of enterocytes and the canalicular membrane of hepatocytes. It functions as a sterol transporter to mediate intestinal cholesterol absorption and counterbalances hepatobiliary cholesterol excretion. NPC1L1 is the molecular target of ezetimibe, a potent cholesterol absorption inhibitor that is widely used in treating hypercholesterolemia. Recent findings suggest that NPC1L1 deficiency or ezetimibe treatment also prevents diet-induced hepatic steatosis and obesity in addition to reducing blood cholesterol. Future studies should focus on molecular mechanisms underlying NPC1L1-dependent cholesterol transport and elucidation of how a cholesterol transporter modulates the pathogenesis of metabolic diseases. PMID:20809793

  17. Sentinel-1 data exploitation for geohazard activity map generation

    NASA Astrophysics Data System (ADS)

    Barra, Anna; Solari, Lorenzo; Béjar-Pizarro, Marta; Monserrat, Oriol; Herrera, Gerardo; Bianchini, Silvia; Crosetto, Michele; María Mateos, Rosa; Sarro, Roberto; Moretti, Sandro

    2017-04-01

    This work is focused on geohazard mapping and monitoring by exploiting Sentinel-1 (A and B) data and the DInSAR (Differential interferometric SAR (Synthetic Aperture Radar)) techniques. Sometimes the interpretation of the DInSAR derived product (like the velocity map) can be complex, mostly for a final user who do not usually works with radar. The aim of this work is to generate, in a rapid way, a clear product to be easily exploited by the authorities in the geohazard management: intervention planning and prevention activities. Specifically, the presented methodology has been developed in the framework of the European project SAFETY, which is aimed at providing Civil Protection Authorities (CPA) with the capability of periodically evaluating and assessing the potential impact of geohazards (volcanic activity, earthquakes, landslides and subsidence) on urban areas. The methodology has three phases, the interferograms generation, the activity map generation, in terms of velocity and accumulated deformation (with time-series), and the Active Deformation Area (ADA) map generation. The last one is the final product, derived from the original activity map by analyzing the data in a Geographic Information System (GIS) environment, which isolate only the true deformation areas over the noise. This product can be more easily read by the authorities than the original activity map, i.e. can be better exploited to integrate other information and analysis. This product also permit an easy monitoring of the active areas.

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

    USDA-ARS?s Scientific Manuscript database

    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...

  19. 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

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

    PubMed

    Northoff, Georg

    2017-09-01

    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). Copyright © 2017 Elsevier Inc. All rights reserved.

  1. 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.

  2. 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.

  3. 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…

  4. 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.

  5. New generation non-stationary portable neutron generators for biophysical applications of Neutron Activation Analysis.

    PubMed

    Marchese, N; Cannuli, A; Caccamo, M T; Pace, C

    2017-01-01

    Neutron sources are increasingly employed in a wide range of research fields. For some specific purposes an alternative to existing large-scale neutron scattering facilities, can be offered by the new generation of portable neutron devices. This review reports an overview for such recently available neutron generators mainly addressed to biophysics applications with specific reference to portable non-stationary neutron generators applied in Neutron Activation Analysis (NAA). The review reports a description of a typical portable neutron generator set-up addressed to biophysics applications. New generation portable neutron devices, for some specific applications, can constitute an alternative to existing large-scale neutron scattering facilities. Deuterium-Deuterium pulsed neutron sources able to generate 2.5MeV neutrons, with a neutron yield of 1.0×10(6)n/s, a pulse rate of 250Hz to 20kHz and a duty factor varying from 5% to 100%, when combined with solid-state photon detectors, show that this kind of compact devices allow rapid and user-friendly elemental analysis. "This article is part of a Special Issue entitled "Science for Life" Guest Editor: Dr. Austen Angell, Dr. Salvatore Magazù and Dr. Federica Migliardo". Copyright © 2016 Elsevier B.V. All rights reserved.

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

    DTIC Science & Technology

    2016-09-07

    A single residue in Ebola virus receptor NPC1 influences cellular host range 1   in reptiles 2   3   4   Esther Ndungo1, Andrew S. Herbert2...Chandran#, email: kartik.chandran@einstein.yu.edu 19   John M. Dye#, email: john.m.dye1.civ@mail.mil 20   21   Running title: Ebola virus host range...number of disease outbreaks in human 30   populations, including the current unprecedented Ebola virus disease (EVD) outbreak in Western 31

  7. Bisphenol A promotes cholesterol absorption in Caco-2 cells by up-regulation of NPC1L1 expression.

    PubMed

    Feng, Dan; Zou, Jun; Zhang, Shanshan; Li, Xuechun; Li, Peiyang; Lu, Minqi

    2017-01-06

    Bisphenol A (BPA), an commonly exposed environmental chemicals in humans, has been shown to have a hypercholesterolemic effect with molecular mechanism not clear. Since intestinal cholesterol absorption plays a major role in maintaining total body cholesterol homeostasis, the present study is to investigate whether BPA affects cholesterol absorption in the intestinal Caco-2 cells. The Caco-2 cells were pretreated with BPA at different concentrations for 24 h and then incubated with radioactive micellar cholesterol for 2 h. The absorption of radioactive cholesterol was quantified by liquid scintillation. The expression of Niemann-Pick C1-like 1 (NPC1L1) and sterol regulatory element binding protein-2 (SREBP-2) was analyzed by Western blot and qPCR. We found that confluent Caco-2 cells expressed NPC1L1, and the absorption of cholesterol in the cells was inhibited by ezetimibe, a specific inhibitor of NPC1L1. We then pretreated the cells with 0.1-10 nM BPA for 24 h and found that BPA at 1 and 10 nM doses promoted cholesterol absorption. In addition, we found that the BPA-induced promotion of cholesterol absorption was associated with significant increase in the levels of NPC1L1 protein and NPC1L1 mRNA. Moreover, the stimulatory effects of BPA on cholesterol absorption and NPC1L1 expression could be prevented by blockade of the SREBP-2 pathway. This study provides the first evidence that BPA promotes cholesterol absorption in the intestinal cells and the stimulatory effect of BPA is mediated, at least in part, by SREBP-2-NPC1L1 signaling pathway.

  8. The mobile nucleoporin Nup2p and chromatin-bound Prp20p function in endogenous NPC-mediated transcriptional control

    PubMed Central

    Dilworth, David J.; Tackett, Alan J.; Rogers, Richard S.; Yi, Eugene C.; Christmas, Rowan H.; Smith, Jennifer J.; Siegel, Andrew F.; Chait, Brian T.; Wozniak, Richard W.; Aitchison, John D.

    2005-01-01

    Nuclear pore complexes (NPCs) govern macromolecular transport between the nucleus and cytoplasm and serve as key positional markers within the nucleus. Several protein components of yeast NPCs have been implicated in the epigenetic control of gene expression. Among these, Nup2p is unique as it transiently associates with NPCs and, when artificially tethered to DNA, can prevent the spread of transcriptional activation or repression between flanking genes, a function termed boundary activity. To understand this function of Nup2p, we investigated the interactions of Nup2p with other proteins and with DNA using immunopurifications coupled with mass spectrometry and microarray analyses. These data combined with functional assays of boundary activity and epigenetic variegation suggest that Nup2p and the Ran guanylyl-nucleotide exchange factor, Prp20p, interact at specific chromatin regions and enable the NPC to play an active role in chromatin organization by facilitating the transition of chromatin between activity states. PMID:16365162

  9. 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.

  10. The Distribution of Active Force Generators Controls Mitotic Spindle Position

    NASA Astrophysics Data System (ADS)

    Grill, Stephan W.; Howard, Jonathon; Schäffer, Erik; Stelzer, Ernst H. K.; Hyman, Anthony A.

    2003-07-01

    During unequal cell divisions a mitotic spindle is eccentrically positioned before cell cleavage. To determine the basis of the net force imbalance that causes spindle displacement in one-cell Caenorhabditis elegans embryos, we fragmented centrosomes with an ultraviolet laser. Analysis of the mean and variance of fragment speeds suggests that the force imbalance is due to a larger number of force generators pulling on astral microtubules of the posterior aster relative to the anterior aster. Moreover, activation of heterotrimeric guanine nucleotide-binding protein (G protein) α subunits is required to generate these astral forces.

  11. 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

  12. 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.

  13. 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.

  14. Active Interrogation Using Electronic Neutron Generators for Nuclear Safeguards Applications

    SciTech Connect

    David L. Chichester; Edward H. Seabury

    2008-08-01

    Active interrogation, a measurement technique which uses a radiation source to probe materials and generate unique signatures useful for characterizing those materials, is a powerful tool for assaying special nuclear material. The most commonly used technique for performing active interrogation is to use an electronic neutron generator as the probe radiation source. Exploiting the unique operating characteristics of these devices, including their monoenergetic neutron emissions and their ability to operate in pulsed modes, presents a number of options for performing prompt and delayed signature analyses using both photon and neutron sensors. A review of literature in this area shows multiple applications of the active neutron interrogation technique for performing nuclear nonproliferation measurements. Some examples include measuring the plutonium content of spent fuel, assaying plutonium residue in spent fuel hull claddings, assaying plutonium in aqueous fuel reprocessing process streams, and assaying nuclear fuel reprocessing facility waste streams to detect and quantify fissile material. This paper discusses the historical use of this technique and examines its context within the scope and challenges of next-generation nuclear fuel cycles and advanced concept nuclear fuel cycle facilities.

  15. 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.

  16. 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.

  17. Micrurus snake venoms activate human complement system and generate anaphylatoxins

    PubMed Central

    2012-01-01

    Background The genus Micrurus, coral snakes (Serpentes, Elapidae), comprises more than 120 species and subspecies distributed from the south United States to the south of South America. Micrurus snake bites can cause death by muscle paralysis and further respiratory arrest within a few hours after envenomation. Clinical observations show mainly neurotoxic symptoms, although other biological activities have also been experimentally observed, including cardiotoxicity, hemolysis, edema and myotoxicity. Results In the present study we have investigated the action of venoms from seven species of snakes from the genus Micrurus on the complement system in in vitro studies. Several of the Micrurus species could consume the classical and/or the lectin pathways, but not the alternative pathway, and C3a, C4a and C5a were generated in sera treated with the venoms as result of this complement activation. Micrurus venoms were also able to directly cleave the α chain of the component C3, but not of the C4, which was inhibited by 1,10 Phenanthroline, suggesting the presence of a C3α chain specific metalloprotease in Micrurus spp venoms. Furthermore, complement activation was in part associated with the cleavage of C1-Inhibitor by protease(s) present in the venoms, which disrupts complement activation control. Conclusion Micrurus venoms can activate the complement system, generating a significant amount of anaphylatoxins, which may assist due to their vasodilatory effects, to enhance the spreading of other venom components during the envenomation process. PMID:22248157

  18. Interaction between miR-572 and PPP2R2C and their effects on proliferation, migration and invasion of nasopharyngeal carcinoma (NPC) cells.

    PubMed

    Yan, Lei; Cai, Kerui; Liang, Jun; Liu, Haifeng; Liu, Yang; Gui, Jinqiu

    2017-05-19

    We investigated the regulation of miR-572 on PPP2R2C and studied their effects on proliferation, migration as well as invasion of nasopharyngeal carcinoma (NPC) cells. NPC tissues and normal tissues were collected, and the expressions of miR-572 and PPP2R2C were detected by RT-PCR. And Western blot was applied to detect the expression of PPP2R2C protein. The target relationship between miR-572 and PPP2R2C was confirmed by dual luciferase reporter gene assay. MTT assay and flow cytometry was applied to investigate the viability and apoptosis of NPC cells. Transwell as well as wound healing assays were used to respectively detect the invasion and migration ability of NPC cells. MiR-572 was high expressed in NPC tissues as well as NPC cells, and PPP2R2C was low expressed in NPC tissues than normal samples. MiR-572 could bind to 3' UTR of PPP2R2C and decrease its expression. Over-expressed miR-572 and decreased PPP2R2C expression could both inhibit proliferation, invasion and induce apoptosis of NPC cells. MiR-572 could promote the proliferation and invasion of NPC through down-regulating PPP2R2C directly.

  19. 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.

  20. Long Noncoding RNA Highly Upregulated in Liver Cancer Activates p53-p21 Pathway and Promotes Nasopharyngeal Carcinoma Cell Growth.

    PubMed

    Jiang, Xue; Liu, Weiwei

    2017-07-01

    Dysfunction of lncRNA has been found in the nasopharyngeal carcinoma (NPC); however, the effect of lncRNA expression on NPC tumorigenesis as well as the molecular mechanism of lncRNA in the pathogenesis of NPC remain largely unknown. In this study, we showed that highly upregulated in liver cancer (HULC), the first identified lncRNA in hepatocellular carcinoma, is highly expressed in NPC patients and correlated with a poor prognosis of cancer patients. Overexpressed HULC promotes NPC cell growth. Downregulation of HULC activated p53 and induced the increased expression of p21, which finally caused cell cycle arrest and cell apoptosis. These results suggested that HULC acts as an oncogenic lncRNA in NPC and a potential therapeutic target in NPC treatment.

  1. 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

  2. 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

  3. 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

  4. Intrinsically-generated fluctuating activity in excitatory-inhibitory networks.

    PubMed

    Mastrogiuseppe, Francesca; Ostojic, Srdjan

    2017-04-01

    Recurrent networks of non-linear units display a variety of dynamical regimes depending on the structure of their synaptic connectivity. A particularly remarkable phenomenon is the appearance of strongly fluctuating, chaotic activity in networks of deterministic, but randomly connected rate units. How this type of intrinsically generated fluctuations appears in more realistic networks of spiking neurons has been a long standing question. To ease the comparison between rate and spiking networks, recent works investigated the dynamical regimes of randomly-connected rate networks with segregated excitatory and inhibitory populations, and firing rates constrained to be positive. These works derived general dynamical mean field (DMF) equations describing the fluctuating dynamics, but solved these equations only in the case of purely inhibitory networks. Using a simplified excitatory-inhibitory architecture in which DMF equations are more easily tractable, here we show that the presence of excitation qualitatively modifies the fluctuating activity compared to purely inhibitory networks. In presence of excitation, intrinsically generated fluctuations induce a strong increase in mean firing rates, a phenomenon that is much weaker in purely inhibitory networks. Excitation moreover induces two different fluctuating regimes: for moderate overall coupling, recurrent inhibition is sufficient to stabilize fluctuations; for strong coupling, firing rates are stabilized solely by the upper bound imposed on activity, even if inhibition is stronger than excitation. These results extend to more general network architectures, and to rate networks receiving noisy inputs mimicking spiking activity. Finally, we show that signatures of the second dynamical regime appear in networks of integrate-and-fire neurons.

  5. 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.

  6. 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.

  7. 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.

  8. 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.

  9. 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

  10. 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.

  11. 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.

  12. NPC1 defect results in abnormal platelet formation and function: studies in Niemann-Pick disease type C1 patients and zebrafish.

    PubMed

    Louwette, Sophie; Régal, Luc; Wittevrongel, Christine; Thys, Chantal; Vandeweeghde, Gwenny; Decuyper, Elisa; Leemans, Peter; De Vos, Rita; Van Geet, Chris; Jaeken, Jaak; Freson, Kathleen

    2013-01-01

    Niemann-Pick type C is a lysosomal storage disease associated with mutations in NPC1 or NPC2, resulting in an accumulation of cholesterol in the endosomal-lysosomal system. Niemann-Pick type C has a clinical spectrum that ranges from a neonatal rapidly fatal disorder to an adult-onset chronic neurodegenerative disease combined with remarkably, in some cases, hematological defects such as thrombocytopenia, anemia and petechial rash. A role of NPC1 in hematopoiesis was never shown. Here, we describe platelet function abnormalities in three unrelated patients with a proven genetic and biochemical NPC1 defect. Their platelets have reduced aggregations, P-selectin expression and ATP secretions that are compatible with the observed abnormal alpha and reduced dense granules as studied by electron microscopy and CD63 staining after platelet spreading. Their blood counts were normal. NPC1 expression was shown in platelets and megakaryocytes (MKs). In vitro differentiated MKs from NPC1 patients exhibit hyperproliferation of immature MKs with different CD63(+) granules and abnormal cellular accumulation of cholesterol as shown by filipin stainings. The role of NPC1 in megakaryopoiesis was further studied using zebrafish with GFP-labeled thrombocytes or DsRed-labeled erythrocytes. NPC1 depletion in zebrafish resulted in increased cell death in the brain and abnormal cellular accumulation of filipin. NPC1-depleted embryos presented with thrombocytopenia and mild anemia as studied by flow cytometry and real-time QPCR for specific blood cell markers. In conclusion, this is the first report, showing a role of NPC1 in platelet function and formation but further studies are needed to define how cholesterol storage interferes with these processes.

  13. 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

  14. 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.

  15. 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.

  16. 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.

  17. Stat3 inhibitor Stattic exhibits potent antitumor activity and induces chemo- and radio-sensitivity in nasopharyngeal carcinoma.

    PubMed

    Pan, Yunbao; Zhou, Fuling; Zhang, Ronghua; Claret, Francois X

    2013-01-01

    Nasopharyngeal carcinoma (NPC) is an Epstein-Barr virus-associated malignancy most common in East Asia, Africa and Alaska. Radiotherapy and cisplatin-based chemotherapy are the main treatment options. Unfortunately, disease response to concurrent chemoradiotherapy varies among patients with NPC, and many cases are resistant to cisplatin and radiotherapy. Signal transducer and activator of transcription 3 (Stat3) has been implicated in the development and progression of various solid tumors. In this study, we assessed the activation and expression of Stat3 in NPC cells. We found that Stat3 was activated and could be blocked by the small molecule inhibitor Stattic. The inhibition of Stat3 in NPC cells by Stattic decreased the expression of cyclin D1 in a dose- and time-dependent manner. Thus, Stattic was used to target Stat3 in NPC cell lines. We found that Stattic could inhibit cell viability and proliferation in NPC cells and significantly induced apoptosis. Additionally, Stat3 transfection attenuated, whereas Stat3 knockdown enhanced, the effects of Stattic upon cell viability inhibition and apoptosis induction. Furthermore, Stattic sensitized NPC cells to cisplatin and ionizing radiation (IR) by preventing cell proliferation and inducing apoptosis. Taken together, Stattic inhibit Stat3 and display antitumor effect in NPC, and enhanced chemosensitivity and radiosensitivity in NPC. Therefore, our findings provide the base for more rational approaches to treat NPC in the clinic.

  18. 77 FR 16222 - Agency Information Collection Activities; Proposed Collection; Comment Request; Generator...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-03-20

    ... AGENCY Agency Information Collection Activities; Proposed Collection; Comment Request; Generator... this action are private as well as State, Local, or Tribal Governments. Title: Generator Standards... Agency (EPA) has finalized an alternative set of generator requirements applicable to laboratories owned...

  19. Improved catalytic activity of laser generated bimetallic and trimetallic nanoparticles.

    PubMed

    Singh, Rina; Soni, R K

    2014-09-01

    We report synthesis of silver nanoparticles, bimetallic (Al2O3@Ag) nanoparticles and trimetallic (Al2O3@AgAu) nanoparticles by nanosecond pulse laser ablation (PLA) in deionized water. Two-step laser ablation methodologies were adopted for the synthesis of bi- and tri-metallic nanoparticles. In this method a silver or gold target was ablated in colloidal solution of γ-alumina nanoparticles prepared by PLA. The TEM image analysis of bimetallic and trimetallic particles reveals deposition of fine silver particles and Ag-Au alloy particles, respectively, on large alumina particles. The laser generated nanoparticles were tested for catalytic reduction of 4-nitrophenol to 4-aminophenol and showed excellent catalytic behaviour. The catalytic rate was greatly improved by incorporation of additional metal in silver nanoparticles. The catalytic efficiency of trimetallic Al2O3@AgAu for reduction of 4-nitrophenol to 4-aminophenol was remarkably enhanced and the catalytic reaction was completed in just 5 sec. Even at very low concentration, both Al2O3@Ag nanoparticles and Al2O3@AgAu nanoparticles showed improved rate of catalytic reduction than monometallic silver nanoparticles. Our results demonstrate that alumina particles in the solution not only provide the active sites for particle dispersion but also improve the catalytic activity.

  20. 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.

  1. 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.

  2. Detection of nasopharyngeal carcinoma susceptibility with single nucleotide polymorphism analysis using next-generation sequencing technology

    PubMed Central

    Wu, Mu-Yun; Huang, Shu-Jing; Yang, Fan; Qin, Xin-Tian; Liu, Dong; Ding, Ying; Yang, Shu; Wang, Xi-Cheng

    2017-01-01

    Nasopharyngeal carcinoma (NPC) is a head and neck cancer with high incidence in South China and East Asia. To provide a theoretical basis for NPC risk screening and early prevention, we conducted a meta-analysis of relevant literature on the association of single nucleotide polymorphisms (SNP)s with NPC susceptibility. Further, expression of 15 candidate SNPs identified in the meta-analysis was evaluated in a cohort of NPC patients and healthy volunteers using next-generation sequencing technology. Among the 15 SNPs detected in the meta-analysis, miR-146a (rs2910164, C>G), HCG9 (rs3869062, A>G), HCG9 (rs16896923, T>C), MMP2 (rs243865, C>T), GABBR1 (rs2076483, T>C), and TP53 (rs1042522, C>G) were associated with decreased susceptibility to NPC, while GSTM1 (+/DEL), IL-10 (rs1800896, A>G), MDM2 (rs2279744, T>G), MDS1-EVI1 (rs6774494, G>A), XPC (rs2228000, C>T), HLA-F (rs3129055, T>C), SPLUNC1 (rs2752903, T>C; and rs750064, A>G), and GABBR1 (rs29232, G>A) were associated with increased susceptibility to NPC. In our case-control study, an association with increased risk for NPC was found for the AG vs AA genotype in HCG9 (rs3869062, A>G). In addition, heterozygous deletion of the GSTM1 allele was associated with increased susceptibility to NPC, while an SNP in GABBR1 (rs29232, G>A) was associated with decreased risk, and might thus have a protective role on NPC carcinogenesis. This work provides the first comprehensive assessment of SNP expression and its relationship to NPC risk. It suggests the need for well-designed, larger confirmatory studies to validate its findings. PMID:28881764

  3. 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.

  4. Women's income generation activities in Merowe Province, Northern State, Sudan.

    PubMed

    Pitamber, S; Osama, S

    1994-06-01

    Merowe province in rural northern Sudan has been divided into three local government council areas: Merowe, Karima, and Ed Debba. A government program was instituted to increase the welfare of residents and food production. A baseline survey of 490 respondents was conducted in order to ascertain how illiterate women viewed development in the area and to provide useful information for program design and implementation. Women from 24 villages were administered questionnaires, observed in their daily activities, and engaged in discussion in a local meeting place. Discussions were also held with members of the local Popular Committee. Demographic information was very sketchy about age, and 48% had no formal education in writing and reading. General reading and writing skills of the remainder were very poor. There were 500 female children and 502 male children, and the sex ratio varied among the 3 council areas. 52% were married and 14% were divorced or widowed and living with relatives. The average monthly income was from Ls. 700 to Ls. 3000 based on reports from only 59.3% of respondents. Most of the women had skills in food processing and 25.7% were skilled in handicrafts. Water was obtained primarily from local wells and not decontaminated before use. Pit latrines were the standard. One bathing facility was available in the compound for the entire council area. Health units were either in each village or within 20-30 minutes walk. Child mortality was 4.3% in Merowe province. 77 children 0-5 years old died out of a total of 1002 live births. Life expectancy was 41-50 years for women and 61-70 years for men. Cleanliness and healthful eating were observed. 58% owned no land; plots were under 5 feddans and usually half a feddan. 92.1% had no bank account and 90% had no experience with loans. 70.2% were indifferent about involvement in an income generation program. 26% were interested in part-time participation. Only 3.9% desired full-time participation. 8.6% said they

  5. NPC2, the protein deficient in Niemann-Pick C2 disease, consists of multiple glycoforms that bind a variety of sterols.

    PubMed

    Liou, Heng-Ling; Dixit, Sayali S; Xu, Sujuan; Tint, G Stephen; Stock, Ann M; Lobel, Peter

    2006-12-01

    Niemann-Pick C disease is a fatal neurodegenerative disorder characterized by an endolysosomal accumulation of cholesterol and other lipids. One form of the disease is caused by a deficiency in NPC2, a soluble lysosomal glycoprotein that binds cholesterol. To better understand the biological function of NPC2 and how its deficiency results in disease, we have characterized the structural and functional properties of recombinant human protein. Highly purified NPC2 consists of a complex mixture of glycosylated isoforms, similar to that observed in human brain autopsy specimens. Mass spectrometric analysis revealed that of the three potential N-linked glycosylation sites present in the mature protein, Asn-19 is not utilized; Asn-39 is linked to an endoglycosidase H (Endo H)-sensitive oligosaccharide, and Asn-116 is variably utilized, either being unmodified or linked to Endo H-sensitive or Endo H-resistant oligosaccharides. All glycoforms are endocytosed and ameliorate the cholesterol storage phenotype of NPC2-deficient fibroblasts. In addition, the purified preparation contains a mixture of both free and lipid-bound protein. All glycoforms bind cholesterol, and sterol binding to NPC2 significantly alters its behavior upon cation-exchange chromatography. Based on this observation, we developed chromatography-based binding assays and determined that NPC2 forms an equimolar complex with the fluorescent cholesterol analog dehydroergosterol. In addition, we find that NPC2 binds a range of cholesterol-related molecules (cholesterol precursors, plant sterols, some oxysterols, cholesterol sulfate, cholesterol acetate, and 5-alpha-cholestan-3-one) and that 27-hydroxysterol accumulates in NPC2-deficient mouse liver. Binding was not detected for various glycolipids, phospholipids, or fatty acids. These biochemical properties support a direct and specialized function of NPC2 in lysosomal sterol transport.

  6. 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

    2017-02-01

    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. Copyright © 2016 The Authors. Published by Elsevier Inc. All rights reserved.

  7. 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.

  8. 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.

  9. 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.

  10. 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.

  11. 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.

  12. Prevention of oxLDL uptake leads to decreased atherosclerosis in hematopoietic NPC1-deficient Ldlr(-/-) mice.

    PubMed

    Jeurissen, Mike L J; Walenbergh, Sofie M A; Houben, Tom; Gijbels, Marion J J; Li, Jieyi; Hendrikx, Tim; Oligschlaeger, Yvonne; van Gorp, Patrick J; Binder, Christoph J; Donners, Marjo M P C; Shiri-Sverdlov, Ronit

    2016-12-01

    Atherosclerosis is a chronic inflammatory disease of medium and large vessels and is typically characterized by the predominant accumulation of low-density lipoprotein (LDL)-cholesterol inside macrophages that reside in the vessel walls. Previous studies clearly demonstrated an association specifically between the oxidized type of LDL (oxLDL) and atherosclerotic lesion formation. Further observations revealed that these atherosclerotic lesions displayed enlarged, lipid-loaded lysosomes. By increasing natural antibodies against oxLDL, pneumococcal vaccination has been shown to reduce atherosclerosis in LDL receptor knockout (Ldlr(-/-)) mice. Relevantly, loss of the lysosomal membrane protein Niemann-Pick Type C1 (NPC1) led to lysosomal accumulation of various lipids and promoted atherosclerosis. Yet, the importance of lysosomal oxLDL accumulation inside macrophages, compared to non-modified LDL, in atherosclerosis has never been established. By transplanting NPC1 bone marrow into lethally irradiated Ldlr(-/-) mice, a hematopoietic mouse model for lysosomal cholesterol accumulation was created. Through injections with heat-inactivated pneumococci, we aimed to demonstrate the specific contribution of lysosomal oxLDL accumulation inside macrophages in atherosclerosis development. While there were no differences in plaque morphology, a reduction in plaque size and plaque inflammation was found in immunized NPC1(mut)-transplanted mice, compared to non-immunized NPC1(mut)-transplanted mice. Lysosomal oxLDL accumulation within macrophages contributes to murine atherosclerosis. Future intervention strategies should focus specifically on preventing oxLDL, unlike non-modified LDL, from being internalized into lysosomes. Such an intervention can have an additive effect to current existing treatments against atherosclerosis. Copyright © 2016 The Authors. Published by Elsevier Ireland Ltd.. All rights reserved.

  13. 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

  14. 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

  15. 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.

  16. 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

  17. 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…

  18. 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…

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

    PubMed

    Berle, Jan Øystein; Løberg, Else-Marie; Fasmer, Ole Bernt

    2013-05-06

    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. 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. 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.

  20. 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…

  1. 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…

  2. An Analysis of Generational Differences Among Active Duty Members

    DTIC Science & Technology

    2004-03-01

    studied differences between Baby Boomers (born 1946 – 1964), Generation Xers (born 1965 – 1977), and Millenials (born 1978 – 1995). They found that work...1946-1964) Generation X (1965-1978) Millenials (1979-1994) Cufaude (2000) Matures (1909-1945...Mean & Standard Deviation Comparison Smola & Sutton (2002) Study Sample (2003) 2003 Survey Items Boomers Gen-X Boomers Gen-X Millenials

  3. Early Hippocampal i-LTP and LOX-1 Overexpression Induced by Anoxia: A Potential Role in Neurodegeneration in NPC Mouse Model.

    PubMed

    Lo Castro, Adriana; Murdocca, Michela; Pucci, Sabina; Zaratti, Anna; Greggi, Chiara; Sangiuolo, Federica; Tancredi, Virginia; Frank, Claudio; D'Arcangelo, Giovanna

    2017-07-05

    Niemann-Pick type C disease (NPCD) is an autosomal recessive storage disorder, characterized by abnormal sequestration of unesterified cholesterol within the late endo-lysosomal compartment of cells. In the central nervous system, hypoxic insults could result in low-density lipoprotein (LDL) oxidation and Lectin-like oxidized LDL receptor-1 (LOX-1) induction, leading to a pathological hippocampal response, namely, ischemic long-term potentiation (i-LTP). These events may correlate with the progressive neural loss observed in NPCD. To test these hypotheses, hippocampal slices from Wild Type (WT) and NPC1(-/-) mice were prepared, and field potential in the CA1 region was analyzed during transient oxygen/glucose deprivation (OGD). Moreover, LOX-1 expression was evaluated by RT-qPCR, immunocytochemical, and Western blot analyses before and after an anoxic episode. Our results demonstrate the development of a precocious i-LTP in NPC1(-/-) mice during OGD application. We also observed a higher expression of LOX-1 transcript and protein in NPC1(-/-) mice with respect to WT mice; after anoxic damage to LOX-1 expression, a further increase in both NPC1(-/-) and WT mice was observed, although the protein expression seems to be delayed, suggesting a different kinetic of induction. These data clearly suggest an elevated susceptibility to neurodegeneration in NPC1(-/-) mice due to oxidative stress. The observed up-regulation of LOX-1 in the hippocampus of NPC1(-/-) mice may also open a new scenario in which new biomarkers can be identified.

  4. 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.

  5. Active electroreception in Gymnotus omari: imaging, object discrimination, and early processing of actively generated signals.

    PubMed

    Caputi, Angel A; Castelló, María E; Aguilera, Pedro A; Pereira, Carolina; Nogueira, Javier; Rodríguez-Cattaneo, Alejo; Lezcano, Carolina

    2008-01-01

    Weakly electric fishes "electrically illuminate" the environment in two forms: pulse fishes emit a succession of discrete electric discharges while wave fishes emit a continuous wave. These strategies are present in both taxonomic groups of weakly electric fishes, mormyrids and gymnotids. As a consequence one can distinguish four major types of active electrosensory strategies evolving in parallel. Pulse gymnotids have an electrolocating strategy common with pulse mormyrids, but brains of pulse and wave gymnotids are alike. The beating strategy associated to other differences in the electrogenic system and electrosensory responses suggests that similar hardware might work in a different mode for processing actively generated electrosensory images. In this review we summarize our findings in pulse gymnotids' active electroreception and outline a primary agenda for the next research.

  6. 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

  7. 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.

  8. 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.

  9. Electrophilic Aldehydes Generated by Sperm Metabolism Activate Mitochondrial Reactive Oxygen Species Generation and Apoptosis by Targeting Succinate Dehydrogenase*

    PubMed Central

    Aitken, R. John; Whiting, Sara; De Iuliis, Geoffry N.; McClymont, Samantha; Mitchell, Lisa A.; Baker, Mark A.

    2012-01-01

    Oxidative stress is a major cause of defective sperm function in cases of male infertility. Such stress is known to be associated with high levels of superoxide production by the sperm mitochondria; however, the causes of this aberrant activity are unknown. Here we show that electrophilic aldehydes such as 4-hydroxynonenal (4HNE) and acrolein, generated as a result of lipid peroxidation, target the mitochondria of human spermatozoa and stimulate mitochondrial superoxide generation in a dose- and time-dependent manner. The activation of mitochondrial electron leakage by 4HNE is shown to involve the disruption of succinate dehydrogenase activity and subsequent activation of an intrinsic apoptotic cascade beginning with a loss of mitochondrial membrane potential and terminating in oxidative DNA adduct formation, DNA strand breakage, and cell death. A tight correlation between spontaneous mitochondrial superoxide generation and 4HNE content (R2 = 0.89) in untreated populations of human spermatozoa emphasized the pathophysiological significance of these findings. The latter also provide a biochemical explanation for the self-perpetuating nature of oxidative stress in the male germ line, with the products of lipid peroxidation stimulating free radical generation by the sperm mitochondria in a positive feedback loop. PMID:22851170

  10. 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

  11. Liposomal Packaging Generates Wnt Protein with In Vivo Biological Activity

    PubMed Central

    Zhao, Ludan; Kim, Jae-Beom; ten Berge, Derk; Ponnusamy, Karthik; Carre, A. Lyonel; Dudek, Henryk; Zachlederova, Marie; McElhaney, Michael; Brunton, Shirley; Gunzner, Janet; Callow, Marinella; Polakis, Paul; Costa, Mike; Zhang, Xiaoyan M.; Helms, Jill A.; Nusse, Roel

    2008-01-01

    Wnt signals exercise strong cell-biological and regenerative effects of considerable therapeutic value. There are, however, no specific Wnt agonists and no method for in vivo delivery of purified Wnt proteins. Wnts contain lipid adducts that are required for activity and we exploited this lipophilicity by packaging purified Wnt3a protein into lipid vesicles. Rather than being encapsulated, Wnts are tethered to the liposomal surface, where they enhance and sustain Wnt signaling in vitro. Molecules that effectively antagonize soluble Wnt3a protein but are ineffective against the Wnt3a signal presented by a cell in a paracrine or autocrine manner are also unable to block liposomal Wnt3a activity, suggesting that liposomal packaging mimics the biological state of active Wnts. When delivered subcutaneously, Wnt3a liposomes induce hair follicle neogenesis, demonstrating their robust biological activity in a regenerative context. PMID:18698373

  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. 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

  14. 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.

  15. 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.

  16. 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

  17. 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.

  18. 20 CFR 627.225 - Employment generating activities.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... Section 627.225 Employees' Benefits EMPLOYMENT AND TRAINING ADMINISTRATION, DEPARTMENT OF LABOR GENERAL... decisions about community job training needs; subscriptions to relevant publications; general dissemination... training (OJT) opportunities, as defined in § 627.240; and other allowable JTPA activities in the private...

  19. Multi-Generational Learning: Developing Informed Advocates and Active Children

    ERIC Educational Resources Information Center

    Carson, Linda M.

    2004-01-01

    The creation of multifaceted, multigenerational programs in college communities could create a ripple effect of advocacy for physical education with students and parents becoming agents for change in their own families, communities, cultures, and professions. This article describes community-based physical activity programs at West Virginia…

  20. 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…

  1. 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…

  2. 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

  3. 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.

  4. Activation of Central Pattern Generator for Respiration Following Complete High Cervical Spinal Cord Interruption

    DTIC Science & Technology

    2016-09-01

    AWARD NUMBER: W81XWH-15-1-0324 TITLE: Activation of Central Pattern Generator for Respiration Following Complete High Cervical Spinal Cord...COVERED 1 Sep 2015 - 31 Aug 2016 4. TITLE AND SUBTITLE Activation of Central Pattern Generator for Respiration Following Complete High Cervical...the control of a spinal central pattern generator (CPG) for breathing. These inhibitory spinal interneurons were hypothesized to contribute to

  5. Control of Hydrogen Generation from Water Molecules Dissociated by Activated Aluminum Particles Based on Fuzzy Logic

    NASA Astrophysics Data System (ADS)

    Maekawa, Koji; Takahara, Kenji; Kajiwara, Toshinori; Watanabe, Masao

    This paper proposes a control system to keep hydrogen generation by a reaction between water and activated aluminum particles at desired level. Because the activated aluminum particles are produced shredded aluminum sawdust, the characteristics of hydrogen generation vary depending on its samples. Therefore, the fuzzy control system to determine the quantum of the activated aluminum particles is designed based on the measured characteristics of hydrogen generation. Error form a desired value, error rate and dead time of the reaction are chosen as the labels of the proposed fuzzy membership functions. The reactor vessel that the activated aluminum particles are put into is developed to generate hydrogen continuously. Three types of aluminum particles of the characteristic are used for the experiments. The proposed system is confirmed to be useful for the control of hydrogen generation, coping with the effect of reacting characteristic changes according to the activated aluminum samples.

  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. A new model predictive control algorithm by reducing the computing time of cost function minimization for NPC inverter in three-phase power grids.

    PubMed

    Taheri, Asghar; Zhalebaghi, Mohammad Hadi

    2017-08-17

    This paper presents a new control strategy based on finite-control-set model-predictive control (FCS-MPC) for Neutral-point-clamped (NPC) three-level converters. Containing some advantages like fast dynamic response, easy inclusion of constraints and simple control loop, makes the FCS-MPC method attractive to use as a switching strategy for converters. However, the large amount of required calculations is a problem in the widespread of this method. In this way, to resolve this problem this paper presents a modified method that effectively reduces the computation load compare with conventional FCS-MPC method and at the same time does not affect on control performance. The proposed method can be used for exchanging power between electrical grid and DC resources by providing active and reactive power compensations. Experiments on three-level converter for three Power Factor Correction (PFC), inductive and capacitive compensation modes verify the good and comparable performance. The results have been simulated using MATLAB/SIMULINK software. Copyright © 2017 ISA. Published by Elsevier Ltd. All rights reserved.

  8. 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.

  9. 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

  10. 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.

  11. 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.

  12. 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

  13. 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.

  14. Synaptically Activated Burst-Generating Conductances Underlie a Group-Pacemaker Mechanism for Respiratory Rhythm Generation in Mammals

    PubMed Central

    Del Negro, Christopher A.; Hayes, John A.; Pace, Ryland W.; Brush, Benjamin R.; Teruyama, Ryoichi; Feldman, Jack L.

    2012-01-01

    Breathing, chewing and walking are critical life-sustaining behaviors in mammals that consist essentially of simple rhythmic movements. Breathing movements in particular involve the diaphragm, thorax, and airways but emanate from a network in the lower brain stem. This network can be studied in reduced preparations in vitro and using simplified mathematical models that make testable predictions. An iterative approach that employs both in vitro and in silico models has ruled out canonical mechanisms for respiratory rhythm that involve reciprocal inhibition and pacemaker properties. We present an alternative model in which emergent network properties play the key rhythmogenic role. Specifically, we show evidence that synaptically activated burst-generating conductances – which are only available in the context of network activity – engender robust periodic bursts in respiratory neurons. Because the cellular burst-generating mechanism is linked to network synaptic drive we dub this type of system a group pacemaker. PMID:21111204

  15. 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

  16. 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).

  17. 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

  18. Generation of radicals and antimalarial activity of dispiro-1,2,4-trioxolanes

    NASA Astrophysics Data System (ADS)

    Denisov, E. T.; Denisova, T. G.

    2013-01-01

    The kinetic schemes of the intramolecular oxidation of radicals generated from substituted dispiro-1,2,4-trioxolanes (seven compounds) in the presence of Fe2+ and oxygen were built. Each radical reaction was defined in terms of enthalpy, activation energy, and rate constant. The kinetic characteristics were calculated by the intersecting parabolas method. The competition between the radical reactions was considered. The entry of radicals generated by each compound into the volume was calculated. High antimalarial activity was found for 1,2,4-trioxolanes, which generated hydroxyl radicals. The structural features of trioxolanes responsible for the generation of hydroxyl radicals were determined.

  19. Early Hippocampal i-LTP and LOX-1 Overexpression Induced by Anoxia: A Potential Role in Neurodegeneration in NPC Mouse Model

    PubMed Central

    Lo Castro, Adriana; Murdocca, Michela; Pucci, Sabina; Greggi, Chiara; Sangiuolo, Federica; Tancredi, Virginia; Frank, Claudio; D’Arcangelo, Giovanna

    2017-01-01

    Niemann-Pick type C disease (NPCD) is an autosomal recessive storage disorder, characterized by abnormal sequestration of unesterified cholesterol within the late endo-lysosomal compartment of cells. In the central nervous system, hypoxic insults could result in low-density lipoprotein (LDL) oxidation and Lectin-like oxidized LDL receptor-1 (LOX-1) induction, leading to a pathological hippocampal response, namely, ischemic long-term potentiation (i-LTP). These events may correlate with the progressive neural loss observed in NPCD. To test these hypotheses, hippocampal slices from Wild Type (WT) and NPC1−/− mice were prepared, and field potential in the CA1 region was analyzed during transient oxygen/glucose deprivation (OGD). Moreover, LOX-1 expression was evaluated by RT-qPCR, immunocytochemical, and Western blot analyses before and after an anoxic episode. Our results demonstrate the development of a precocious i-LTP in NPC1−/− mice during OGD application. We also observed a higher expression of LOX-1 transcript and protein in NPC1−/− mice with respect to WT mice; after anoxic damage to LOX-1 expression, a further increase in both NPC1−/− and WT mice was observed, although the protein expression seems to be delayed, suggesting a different kinetic of induction. These data clearly suggest an elevated susceptibility to neurodegeneration in NPC1−/− mice due to oxidative stress. The observed up-regulation of LOX-1 in the hippocampus of NPC1−/− mice may also open a new scenario in which new biomarkers can be identified. PMID:28678158

  20. Next-generation electroceramic fibers for active control

    NASA Astrophysics Data System (ADS)

    Bystricky, Pavel; Pascucci, Marina R.; Strock, Harold B.

    2002-07-01

    Lead-based PMN-31PT and lead-free BNBZT fibers in the 250- 500 micrometer diameter range were produced using CeraNova's proprietary extrusion technology. Various recrystallization approaches were investigated, including seeded solid state conversion and self-seeded texturing, with the goal of obtaining single-crystalline or textured macrocrystalline fibers. Grains in excess of 100 micrometers - and exceeding 1 mm in some cases - with surface and bulk coverage approaching 100 percent, were obtained in a narrow temperature range and under carefully controlled atmosphere conditions. Large grain growth in BNBZT required the presence of BaSrTiO3 or SrTiO3 seeds and temperatures in the 1150-1200 degrees C range. In PMN-31PT, nearly compete recrystalline was observed in unseeded material at relatively low temperature and short time, and improved performance was achieved with a two-step sintering schedule and slightly extended time. While conduction effects have not yet allowed compete assessment of recrystalline BNBZT, PMN-31PT fibers have shown excellent piezoelectric properties with remanent polarization in excess of 30(mu) C/cm2 and coercive field of 4.5kV/cm. When incorporated into active fiber composites, the latter fibers' performance of 2000 microstrain in superior to average PZT-based production composites. Efforts are under way to induce preferred orientation in the large crystal in order to maximize performance.

  1. 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.

  2. Berberine suppresses tumorigenicity and growth of nasopharyngeal carcinoma cells by inhibiting STAT3 activation induced by tumor associated fibroblasts

    PubMed Central

    2013-01-01

    Background Cortidis rhizoma (Huanglian) and its major therapeutic component, berberine, have drawn extensive attention in recent years for their anti-cancer properties. Growth inhibitory effects of berberine on multiple types of human cancer cells have been reported. Berberine inhibits invasion, induces cell cycle arrest and apoptosis in human cancer cells. The anti-inflammatory property of berberine, involving inhibition of Signal Transducer and Activator of Transcription 3 (STAT3) activation, has also been documented. Methods In this study, we have examined the effects of berberine on tumorigenicity and growth of nasopharyngeal carcinoma (NPC) cells and their relationship to STAT3 signaling using both in vivo and in vitro models. Results Berberine effectively inhibited the tumorigenicity and growth of an EBV-positive NPC cell line (C666-1) in athymic nude mice. Inhibition of tumorigenic growth of NPC cells in vivo was correlated with effective inhibition of STAT3 activation in NPC cells inside the tumor xenografts grown in nude mice. In vitro, berberine inhibited both constitutive and IL-6-induced STAT3 activation in NPC cells. Inhibition of STAT3 activation by berberine induced growth inhibition and apoptotic response in NPC cells. Tumor-associated fibroblasts were found to secret IL-6 and the conditioned medium harvested from the fibroblasts also induced STAT3 activation in NPC cells. Furthermore, STAT3 activation by conditioned medium of tumor-associated fibroblasts could be blocked by berberine or antibodies against IL-6 and IL-6R. Conclusions Our observation that berberine effectively inhibited activation of STAT3 induced by tumor-associated fibroblasts suggests a role of berberine in modulating the effects of tumor stroma on the growth of NPC cells. The effective inhibition of STAT3 activation in NPC cells by berberine supports its potential use in the treatment of NPC. PMID:24380387

  3. 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

  4. Comparison of markers of coagulation activation and thrombin generation test in uncomplicated pregnancies.

    PubMed

    Joly, Berangere; Barbay, Virginie; Borg, Jeanne-Yvonne; Le Cam-Duchez, Veronique

    2013-09-01

    Pregnancy is a well-established risk factor for venous thromboembolism, and is associated with a state of hypercoagulability or parameters of thrombin generation. Currently, there is a lack of consensual data on thrombin generation during pregnancy. This study aimed to find a sensitive and specific biological marker of coagulation activation and to identify parameters of thrombin generation. The population included 101 women with uncomplicated pregnancies. The objective of this study was to correlate thrombin generation test (measured at 5pM tissue factor, 4μM lipids and without thrombomodulin), with fibrinogen and markers of blood coagulation activation: D-dimer, prothrombin fragments 1+2 (F1+2), thrombin-antithrombin complexes (TAT) and fibrin monomer complexes (FMC) in these women. Internal quality control was performed in each set of experiments. Fibrinogen, D-dimer, F1+2, and TAT concentrations increased significantly throughout pregnancy, and were correlated with term of pregnancy. In our study, thrombin generation seemed to increase early on, and then remained stable throughout normal pregnancy, in contrast with other markers of blood coagulation activation, excepting FMC. The latter are subject to large inter-individual variations, especially during second trimester. No correlation was demonstrated between thrombin generation parameters and other activation markers. While markers of coagulation activation significantly increased during pregnancy, thrombin generation increased only early on and remains stable during pregnancy. Finding a sensitive and specific biological marker for vascular pregnancy complications, such as FMC and thrombin generation levels, requires further investigation. © 2013.

  5. Vesicles generated during storage of red blood cells enhance the generation of radical oxygen species in activated neutrophils.

    PubMed

    Jank, Herbert; Salzer, Ulrich

    2011-01-18

    Erythrocytes are known to shed vesicles in vivo, under various conditions in vitro, and, with impact for transfusion medicine, during storage of red blood cell concentrates (Vsto vesicles). Vsto vesicles of blood transfusions have been shown to deliver glycosylphosphatidylinositol-linked proteins to recipient erythrocytes, to display prothrombotic activity, and to have an inhibitory effect on macrophages. The interaction of Vsto vesicles with and their effect on neutrophilic granulocytes has not yet been studied in detail. Fluorescently labeled Vsto and calcium-induced vesicles were prepared in order to study the uptake of labeled vesicular components by neutrophils as compared to the process of phagocytosis of zymosan using flow cytometry and confocal microscopy. The activating effect of Vsto vesicles on neutrophils was addressed by a luminometric assay for stimulated radical oxygen species (ROS) generation.Coincubation of vesicles and neutrophils results in a transfer of vesicular components to the cells. This uptake is different from a phagocytotic process and is enhanced upon interference with the cellular actin cytoskeleton. Preincubation of neutrophils with Vsto vesicles results in an enhanced ROS generation by neutrophils,which is further increased upon fMLP stimulation and during zymosan phagocytosis. The activating effect of Vsto vesicles on neutrophils might be due to the specific accumulation of lysophospholipids in Vsto vesicles and should be considered as a possible contributor to the pathogenesis of transfusion-related acute lung injury.

  6. 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…

  7. 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…

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

    PubMed

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

    2015-12-15

    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. © The Author 2015. Published by Oxford University Press. All rights reserved. For

  9. 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.

  10. 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

  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. 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.

  13. 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.

  14. Extracellular generation of hydrogen peroxide is responsible for activation of EGF receptor by ultraviolet A radiation.

    PubMed

    von Montfort, Claudia; Fernau, Niklas S; Beier, Juliane I; Sies, Helmut; Klotz, Lars-Oliver

    2006-11-01

    Receptor tyrosine kinases such as the epidermal growth factor receptor (EGFR) have been proposed to be activated in cells exposed to ultraviolet A (UVA) radiation (320-400 nm) and to be involved in photocarcinogenesis. Singlet oxygen and hydrogen peroxide are being discussed as mediators of the activation of signal transduction pathways by UVA. It is demonstrated here that EGFR is not activated in cells exposed to UVA in the absence of extracellular photosensitizers. Rather, UVA was capable of activating the EGFR and the related ErbB2 receptor tyrosine kinase in HeLa cells and human keratinocytes only under conditions that allowed for the extracellular photochemical generation of H(2)O(2), such as when cells were covered with cell culture medium during exposure to UVA. Pretreatment of cells with vanadate was required for UVA-induced EGFR activation, pointing to the involvement of protein tyrosine phosphatases. Unlike H(2)O(2), photochemically generated singlet oxygen did not activate EGFR but instead impaired the activation of EGFR by its ligand, EGF. In summary, extracellularly generated H(2)O(2) mediates UVA-induced activation of the EGFR and of ErbB2, whereas intracellular generation of reactive oxygen species upon exposure of cells to UVA is not sufficient for activation of the receptor.

  15. Contribution of generative leisure activities to cognitive function in elderly Sri Lankan adults.

    PubMed

    Maselko, Joanna; Sebranek, Matthew; Mun, Mirna H; Perera, Bilesha; Ahs, Jill; Ostbye, Truls

    2014-09-01

    To examine the unique contribution of generative leisure activities, defined as activities motivated by a concern for others and a need to contribute something to the next generation. Cross-sectional survey. Peri-urban and rural area in southern Sri Lanka. Community-dwelling adults aged 60 and older (N = 252). The main predictors were leisure activities, grouped into generative, social, or solitary. The main outcome was cognitive function, assessed using the Montreal Cognitive Assessment (MoCA) and the Informant Questionnaire on Cognitive Decline in the Elderly (IQCODE). More-frequent engagement in generative leisure activities was associated with higher levels of cognitive function, independent of the effect of other social and solitary leisure activities. In a fully adjusted model combining all three leisure activities, generative activities independently predicted cognitive function as measured using the MoCA (β = 0.47, 95% confidence interval (CI) = 0.11-0.83) and the IQCODE (β = -0.81, 95% CI = -1.54 to -0.09). In this combined model, solitary activities were also independently associated with slower cognitive decline using the MoCA (β = 0.40, 95% CI = 0.16-0.64) but not the IQCODE (β = -0.38, 95% CI = -0.88-0.12); the association with social activities did not reach statistical significance with either measure. These associations did not differ meaningfully according to sex. Generative leisure activities are a promising area for the development of interventions aimed at reducing cognitive decline in elderly adults. © 2014, Copyright the Authors Journal compilation © 2014, The American Geriatrics Society.

  16. Regulation of H2O2 generation in thyroid cells does not involve Rac1 activation.

    PubMed

    Fortemaison, N; Miot, F; Dumont, J E; Dremier, S

    2005-01-01

    The H2O2 generating system of the thyrocyte and the O2- generating system of macrophages and leukocytes present numerous functional analogies. The main constituent enzymes belong to the NADPH oxidase (NOX) family (Duox/ThOX for the thyroid and NOX2 /gp91phox for the leukocytes and macrophages), and in both cell types, H2O2 generation is activated by the intracellular generation of Ca2+ and diacylglycerol signals. Nevertheless, although the controls involved in these two systems are similar, their mechanisms are different. The main factors controlling O2- production by NOX2 are the cytosolic proteins p67phox and p47phox, and Rac, a small GTP-binding protein. We have previously reported that there is no expression of p67phox and p47phox in thyrocytes. Here, we investigated whether Rac1 is an actor in the thyroid H2O2-generating system. Ionomycin- and carbamylcholine-stimulated H2O2 generation was measured in dog thyroid cells pretreated with the Clostridium difficile toxin B, which inhibits Rac proteins. Activation of Rac1 was measured in response to agents stimulating H2O2 production, using the CRIB domain of PAK1 as a probe in a glutathione S-transferase (GST) pull-down assay. Among the various agents inducing H2O2 generation in dog thyrocytes, carbamylcholine is the only one which activates Rac1, whereas phorbol ester and calcium increase alone have no effect, and cAMP inactivates it. Moreover, whereas toxin B inhibits the stimulation of O2- generation by phorbol ester in leukocytes, it does not inhibit H2O2 generation induced by carbamylcholine and ionomycin in dog thyrocytes. Unlike in leukocytes, Rac proteins do not play a role in H2O2 generation in thyroid cells. A different regulatory cascade for the control of H2O2 generation remains to be defined.

  17. 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…

  18. 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.…

  19. 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.…

  20. Generating Language Activities in Real-Time for English Learners Using Language Muse

    ERIC Educational Resources Information Center

    Burstein, Jill; Madnani, Nitin; Sabatini, John; McCaffrey, Dan; Biggers, Kietha; Dreier, Kelsey

    2017-01-01

    K-12 education standards in the U.S. require all students to read complex texts across many subject areas. The "Language Muse™ Activity Palette" is a web-based language-instruction application that uses NLP algorithms and lexical resources to automatically generate language activities and support English language learners' content…

  1. 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)

  2. 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…

  3. 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…

  4. 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)

  5. 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…

  6. 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…

  7. 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…

  8. 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

  9. 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.

  10. 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.

  11. 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.

  12. Force and power generating mechanism(s) in active muscle as revealed from temperature perturbation studies

    PubMed Central

    Ranatunga, K W

    2010-01-01

    The basic characteristics of the process of force and power generation in active muscle that have emerged from temperature studies are examined. This is done by reviewing complementary findings from temperature-dependence studies and rapid temperature-jump (T-jump) experiments and from intact and skinned fast mammalian muscle fibres. In isometric muscle, a small T-jump leads to a characteristic rise in force showing that crossbridge force generation is endothermic (heat absorbed) and associated with increased entropy (disorder). The sensitivity of the T-jump force generation to added inorganic phosphate (Pi) indicates that a T-jump enhances an early step in the actomyosin (crossbridge) ATPase cycle before Pi-release. During muscle lengthening when steady force is increased, the T-jump force generation is inhibited. Conversely, during shortening when steady force is decreased, the T-jump force generation is enhanced in a velocity-dependent manner, showing that T-jump force generation is strain sensitive. Within the temperature range of ∼5–35°C, the temperature dependence of steady active force is sigmoidal both in isometric and in shortening muscle. However, in shortening muscle, the endothermic character of force generation becomes more pronounced with increased velocity and this can, at least partly, account for the marked increase with warming of the mechanical power output of active muscle. PMID:20660565

  13. 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.

  14. Generation, language, body mass index, and activity patterns in Hispanic children.

    PubMed

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

    2010-02-01

    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. To investigate associations among generation and language with relative weight (BMI z-scores), physical activity, screen time, and participation in extracurricular activities (i.e., sports, clubs) in a U.S.-based, nationally representative sample of Hispanic children. Participants included 2012 Hispanic children aged 6-11 years from the cross-sectional 2003 National Survey of Children's Health. Children were grouped according to generational status (first, second, or third), and the primary language spoken in the home (English versus 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. Compared to third-generation, English speakers, first- and second-generation, non-English speakers were more than two times more likely to be obese. Moreover, first-generation, non-English speakers were half as likely to engage in regular physical activity and sports. Both first- and second-generation, non-English speakers were less likely to participate in clubs compared to second- and third-generation, English speakers. Overall, non-English-speaking groups reported less screen time compared to third-generation, English speakers. 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. Copyright 2010 American Journal of Preventive Medicine. Published by Elsevier Inc. All rights reserved.

  15. 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

  16. Gap Junctions Are Essential for Generating the Correlated Spike Activity of Neighboring Retinal Ganglion Cells

    PubMed Central

    Paul, David L.; Wang, Jack T.; Huberman, Andrew D.; Bloomfield, Stewart A.

    2013-01-01

    Neurons throughout the brain show spike activity that is temporally correlated to that expressed by their neighbors, yet the generating mechanism(s) remains unclear. In the retina, ganglion cells (GCs) show robust, concerted spiking that shapes the information transmitted to central targets. Here we report the synaptic circuits responsible for generating the different types of concerted spiking of GC neighbors in the mouse retina. The most precise concerted spiking was generated by reciprocal electrical coupling of GC neighbors via gap junctions, whereas indirect electrical coupling to a common cohort of amacrine cells generated the correlated activity with medium precision. In contrast, the correlated spiking with the lowest temporal precision was produced by shared synaptic inputs carrying photoreceptor noise. Overall, our results demonstrate that different synaptic circuits generate the discrete types of GC correlated activity. Moreover, our findings expand our understanding of the roles of gap junctions in the retina, showing that they are essential for generating all forms of concerted GC activity transmitted to central brain targets. PMID:23936012

  17. 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.

  18. 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.

  19. 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.

  20. 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

  1. 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.

  2. 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.

  3. 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

  4. Selective optogenetic activation of arcuate kisspeptin neurons generates pulsatile luteinizing hormone secretion

    PubMed Central

    Han, Su Young; McLennan, Timothy; Czieselsky, Katja; Herbison, Allan E.

    2015-01-01

    Normal reproductive functioning in mammals depends upon gonadotropin-releasing hormone (GnRH) neurons generating a pulsatile pattern of gonadotropin secretion. The neural mechanism underlying the episodic release of GnRH is not known, although recent studies have suggested that the kisspeptin neurons located in the arcuate nucleus (ARN) may be involved. In the present experiments we expressed channelrhodopsin (ChR2) in the ARN kisspeptin population to test directly whether synchronous activation of these neurons would generate pulsatile luteinizing hormone (LH) secretion in vivo. Characterization studies showed that this strategy targeted ChR2 to 70% of all ARN kisspeptin neurons and that, in vitro, these neurons were activated by 473-nm blue light with high fidelity up to 30 Hz. In vivo, the optogenetic activation of ARN kisspeptin neurons at 10 and 20 Hz evoked high amplitude, pulse-like increments in LH secretion in anesthetized male mice. Stimulation at 10 Hz for 2 min was sufficient to generate repetitive LH pulses. In diestrous female mice, only 20-Hz activation generated significant increments in LH secretion. In ovariectomized mice, 5-, 10-, and 20-Hz activation of ARN kisspeptin neurons were all found to evoke LH pulses. Part of the sex difference, but not the gonadal steroid dependence, resulted from differential pituitary sensitivity to GnRH. Experiments in kisspeptin receptor-null mice, showed that kisspeptin was the critical neuropeptide underlying the ability of ARN kisspeptin neurons to generate LH pulses. Together these data demonstrate that synchronized activation of the ARN kisspeptin neuronal population generates pulses of LH. PMID:26443858

  5. Inference Generation during Text Comprehension by Adults with Right Hemisphere Brain Damage: Activation Failure Versus Multiple Activation.

    ERIC Educational Resources Information Center

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

    2004-01-01

    ourse comprehensionEvidence 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…

  6. 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.

  7. 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.

  8. 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.

  9. 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.

  10. 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.

  11. The antimicrobial activity of prototype modified honeys that generate reactive oxygen species (ROS) hydrogen peroxide.

    PubMed

    Cooke, Jonathan; Dryden, Matthew; Patton, Thomas; Brennan, James; Barrett, John

    2015-01-28

    Antimicrobial resistance continues to be a global issue in healthcare organisations. Honey has long been shown to possess wound healing and antimicrobial properties that are dependent on a number of physical and chemical properties of the honey. We tested the antimicrobial activity of a medicinal honey, Surgihoney® (SH) and two prototype modified honeys made by Apis mellifera (honeybee) against Staphylococcus aureus (NCIMB 9518). We also examined the modified honey prototypes for the ability to generate reactive oxygen species (ROS) by changing the level of production of hydrogen peroxide from the samples. Surgihoney® (SH) was compared with two modified honeys, Prototype 1 (PT1) and Prototype 2 (PT2) using a bioassay method against a standard strain of Staphylococcus aureus. Further work studied the rate of generation of ROS hydrogen peroxide from these preparations. Surgihoney® antimicrobial activity was shown to be largely due to ROS hydrogen peroxide production. By modification of Surgihoney®, two more potent honey prototypes were shown to generate between a two- and three-fold greater antibacterial activity and up to ten times greater ROS peroxide activity. Surgihoney® is a clinically available wound antiseptic dressing that shows good antimicrobial activity. Two further honey prototypes have been shown to have antimicrobial activity that is possible to be enhanced due to demonstrated increases in ROS peroxide activity.

  12. Lipopolysaccharide (LPS)-mediated macrophage activation: the role of calcium in the generation of tumoricidal activity

    SciTech Connect

    Drysdale, B.E.; Shin, H.S.

    1986-03-01

    As the authors reported, calcium ionophore, A23187, activates macrophages (M theta) for tumor cell killing and the activated M theta produce a soluble cytotoxic factor (M theta-CF) that is similar if not identical to tumor necrosis factor. Based on these observations they have investigated whether calcium is involved in the activation mediated by another potent M theta activator, LPS. The authors have shown that A23187 caused uptake of extracellular /sup 45/Ca/sup + +/ but LPS did not. They have examined the effect of depleting extracellular calcium by using medium containing no added calcium containing 1.0 mM EGTA. In no case did depletion result in decreased M theta-CF production by the M theta activated with LPS. Measurements using the fluorescent, intracellular calcium indicator, Quin 2 have also been performed. While ionomycin, caused a rapid change in the Quin-2 signal, LPS at a concentration even in excess of that required to activate the M theta caused no change in the signal. When high doses of Quin 2 or another intracellular chelator, 8-(diethylaminol-octyl-3,4,5-trimethoxybenzoate, were used to treat M theta, M theta-CF production decreased and cytotoxic activity was impaired. These data indicate that one or more of the processes involved in M theta-CF production does require calcium, but that activation mediated by LPS occurs without the influx of extracellular calcium or redistribution of intracellular calcium.

  13. 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. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  14. 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),…

  15. 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),…

  16. Nuclear envelope breakdown in starfish oocytes proceeds by partial NPC disassembly followed by a rapidly spreading fenestration of nuclear membranes

    PubMed Central

    Lénárt, Péter; Rabut, Gwénaël; Daigle, Nathalie; Hand, Arthur R.; Terasaki, Mark; Ellenberg, Jan

    2003-01-01

    Breakdown of the nuclear envelope (NE) was analyzed in live starfish oocytes using a size series of fluorescently labeled dextrans, membrane dyes, and GFP-tagged proteins of the nuclear pore complex (NPC) and the nuclear lamina. Permeabilization of the nucleus occurred in two sequential phases. In phase I the NE became increasingly permeable for molecules up to ∼40 nm in diameter, concurrent with a loss of peripheral nuclear pore components over a time course of 10 min. The NE remained intact on the ultrastructural level during this time. In phase II the NE was completely permeabilized within 35 s. This rapid permeabilization spread as a wave from one epicenter on the animal half across the nuclear surface and allowed free diffusion of particles up to ∼100 nm in diameter into the nucleus. While the lamina and nuclear membranes appeared intact at the light microscopic level, a fenestration of the NE was clearly visible by electron microscopy in phase II. We conclude that NE breakdown in starfish oocytes is triggered by slow sequential disassembly of the NPCs followed by a rapidly spreading fenestration of the NE caused by the removal of nuclear pores from nuclear membranes still attached to the lamina. PMID:12654902

  17. How to reduce the accumulation of autophagic vacuoles in NPC1-deficient neurons: a comparison of two pharmacological strategies.

    PubMed

    Meske, Volker; Priesnitz, Timm; Albert, Frank; Ohm, Thomas Georg

    2015-02-01

    A disturbed autophagic pathway leads to chronically increased levels of autophagic vacuoles in Niemann Pick Type-C 1 (NPC1) deficient neurons. Since these accumulations potentially contribute to neuronal cell death associated with the disease, we investigated two pharmacological strategies which potentially reduce the number of autophagic structures under following aspects: efficiency, sustainability and effect on neuronal cell viability. The strategies comprised (i) an interruption of the autophagic flux by the class III PI3K inhibitor 3-methyladenine (3-MA) and (ii) an acceleration of the autophagic execution by 2-hydroxypropyl-β-cyclodextrin (pCD). Our data show that the inhibition of autophagy with 3-MA only initially reduced the number of autophagic vacuoles in cultured neurons. Prolonged treatments with the PI3K-inhibitor reversed this lowering effect. The re-increase in the number of autophagic vacuoles was combined with a defect in the integrity of lysosomes which endangered further survival of cells. The treatment with pCD evoked a slow but sustained reduction of autophagic structures and had no negative effects on neuronal survival.

  18. 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.

  19. Simultaneous Active Source Control of Blade Row Interaction Generated Discrete Tones

    NASA Technical Reports Server (NTRS)

    Sawyer, Scott; Fleeter, Sanford

    1999-01-01

    Unsteady blade row interactions in turbomachines generate discrete-frequency tones at blade pass frequency and its harmonics. Specific circumferential acoustic modes are generated. However, only certain of these modes propagate upstream and downstream to the far field, with these the discrete frequency noise received by an observer. This paper is directed at experimentally demonstrating the viability of active noise control utilizing active airfoils to generate propagating spatial modes that interact with and simultaneously cancel the upstream and downstream propagating acoustic modes. This is accomplished by means of fundamental experiments performed in the Purdue Annular Cascade Research Facility configured with 16 rotor blades and 18 stator vanes. At blade pass frequency, only the k(sub 0) = -2 spatial mode propagates. Significant simultaneous noise reductions are achieved for these upstream and downstream propagating spatial modes over a wide range of operating conditions.

  20. 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.

  1. 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.

  2. 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.

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

    USDA-ARS?s Scientific Manuscript database

    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...

  4. 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...

  5. 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...

  6. 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...

  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. 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.

  9. The nucleic acid scavenger polyamidoamine third-generation dendrimer inhibits fibroblast activation and granulation tissue contraction.

    PubMed

    Holl, Eda K; Bond, Jennifer E; Selim, Maria A; Ehanire, Tosan; Sullenger, Bruce; Levinson, Howard

    2014-09-01

    Pathologic cutaneous scarring affects over 40 million people worldwide and costs billions of dollars annually. Understanding mechanisms of fibroblast activation and granulation tissue contraction is the first step toward preventing pathologic scarring. The authors hypothesize that nucleic acids increase fibroblast activation and cause granulation tissue contraction and that sequestration of nucleic acids by application of a nucleic acid scavenger dendrimer, polyamidoamine third-generation dendrimer, will decrease pathologic scarring. In vitro experiments were performed to assess the effect of nucleic acids on pathologic scar-associated fibroblast activity. The effect of nucleic acids on cytokine production and migration on mouse fibroblasts was evaluated. Immunofluorescence microscopy was used to determine the effect of nucleic acids on the differentiation of human primary fibroblasts into myofibroblasts. Using a murine model, the effect of polyamidoamine third-generation dendrimer on granulation tissue contraction was evaluated by gross and histologic parameters. Mouse fibroblasts stimulated with nucleic acids had increased cytokine production (i.e., transforming growth factor-β, monocyte chemotactic protein 1, interleukin-10, tumor necrosis factor-α, and interferon-γ), migration, and differentiation into myofibroblasts. Polyamidoamine third-generation dendrimer blocked cytokine production, migration, and differentiation into myofibroblasts. Using a murine model of granulation tissue contraction, polyamidoamine third-generation dendrimer decreased wound contraction and angiogenesis. Collagen deposition in polyamidoamine third-generation dendrimer-treated tissues was aligned more randomly and whorl-like compared with control tissue. The data demonstrate that nucleic acid-stimulated fibroblast activation and granulation tissue contraction are blocked by polyamidoamine third-generation dendrimer. Sequestration of pathogen-associated molecular patterns may be an

  10. The Nova Scotia (type D) form of Niemann-Pick disease is caused by a G3097-->T transversion in NPC1.

    PubMed

    Greer, W L; Riddell, D C; Gillan, T L; Girouard, G S; Sparrow, S M; Byers, D M; Dobson, M J; Neumann, P E

    1998-07-01

    Niemann-Pick type D (NPD) disease is a progressive neurodegenerative disorder characterized by the accumulation of tissue cholesterol and sphingomyelin. This disorder is relatively common in southwestern Nova Scotia, because of a founder effect. Our previous studies, using classic linkage analysis of this large extended kindred, defined the critical gene region to a 13-cM chromosome segment between D18S40 and D18S66. A recently isolated gene from this region, NPC1, is mutated in the majority of patients with Niemann-Pick type C disease. We have identified a point mutation within this gene (G3097-->T; Gly992-->Trp) that shows complete linkage disequilibrium with NPD, confirming that NPD is an allelic variant of NPC1.

  11. VLDL hydrolysis by LPL activates PPAR-alpha through generation of unbound fatty acids.

    PubMed

    Ruby, Maxwell A; Goldenson, Benjamin; Orasanu, Gabriela; Johnston, Thomas P; Plutzky, Jorge; Krauss, Ronald M

    2010-08-01

    Recent evidence suggests that lipoproteins serve as circulating reservoirs of peroxisomal proliferator activated receptor (PPAR) ligands that are accessible through lipolysis. The present study was conducted to determine the biochemical basis of PPAR-alpha activation by lipolysis products and their contribution to PPAR-alpha function in vivo. PPAR-alpha activation was measured in bovine aortic endothelial cells following treatment with human plasma, VLDL lipolysis products, or oleic acid. While plasma failed to activate PPAR-alpha, oleic acid performed similarly to VLDL lipolysis products. Therefore, fatty acids are likely to be the PPAR-alpha ligands generated by VLDL lipolysis. Indeed, unbound fatty acid concentration determined PPAR-alpha activation regardless of fatty acid source, with PPAR-alpha activation occurring only at unbound fatty acid concentrations that are unachievable under physiological conditions without lipase action. In mice, a synthetic lipase inhibitor (poloxamer-407) attenuated fasting-induced changes in expression of PPAR-alpha target genes. Apolipoprotein CIII (apoCIII), an endogenous inhibitor of lipoprotein and hepatic lipase, regulated access to the lipoprotein pool of PPAR-alpha ligands, because addition of exogenous apoCIII inhibited, and removal of endogenous apoCIII potentiated, lipolytic PPAR-alpha activation. These data suggest that the PPAR-alpha response is generated by unbound fatty acids released locally by lipase activity and not by circulating plasma fatty acids.

  12. Theoretical models of spontaneous activity generation and propagation in the developing retina.

    PubMed

    Godfrey, Keith B; Eglen, Stephen J

    2009-12-01

    Spontaneous neural activity is present in many parts of the developing nervous system, including visual, auditory and motor areas. In the developing retina, nearby neurons are spontaneously active and produce propagating patterns of activity, known as retinal waves. Such activity is thought to instruct the refinement of retinal axons. In this article we review several computational models used to help evaluate the mechanisms that might be responsible for the generation of retinal waves. We then discuss the models relative to the molecular mechanisms underlying wave activity, including gap junctions, neurotransmitters and second messenger systems. We examine how well the models represent these mechanisms and propose areas for future modelling research. The retinal wave models are also discussed in relation to models of spontaneous activity in other areas of the developing nervous system.

  13. 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.

  14. Next generation FIX muteins with FVIII-independent activity for alternative treatment of hemophilia A.

    PubMed

    Quade-Lyssy, P; Abriss, D; Milanov, P; Ungerer, C; Königs, C; Seifried, E; Schüttrumpf, J

    2014-11-01

    FVIII neutralizing antibodies are the main complication of substitution therapy in hemophilia A (HA); auto-antibodies against FVIII causing acquired HA can also occur. Treatment of inhibitor patients remains challenging because prophylactic treatment with existing FVIII bypassing agents, all based on constitutively active coagulation factors, is difficult due to their short half-life. To generate zymogenic FIX variants with FVIII-independent activity for gene- and protein-based therapy for HA. Modifications were introduced into FIX based on current knowledge of FIX structure and FVIII-independent function followed by random screening. Activity, thrombin generation and FX activation by FIX mutants were characterized in the presence and absence of FVIII. Phenotype correction of promising candidates was assessed by the tail-clip assay in FVIII-knockout mice. About 1600 clones were screened and three mutations (L6F, S102N and E185D) identified, which improved FVIII-independent activity in combination with our previously described variant FIX-ITV. By systematic combination of all mutations, six FIX mutants with the desired bypassing activity were designed. Candidate mutants FIX-IDAV and FIX-FIAV demonstrated the most efficient thrombin generation in FVIII-deficient plasma and had considerably increased activities towards FX in the absence of FVIII, in that they showed an up to 5-fold increase in catalytic efficiency. Expression of FIX-IDAV in FVIII knockout mice reduced blood loss after the tail-clip assay, even in the presence of neutralizing FVIII antibodies. Activatable bioengineered FIX molecules (as opposed to pre-activated coagulation factors) with FVIII-independent activity might be a promising tool for improving HA treatment, especially for patients with inhibitors. © 2014 International Society on Thrombosis and Haemostasis.

  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. 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.

  17. 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.

  18. Sphingomyelinase D Activity in Model Membranes: Structural Effects of in situ Generation of Ceramide-1-Phosphate

    PubMed Central

    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. PMID:22558302

  19. 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.

  20. 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.

  1. 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.

  2. Increase of reactive oxygen species generation in cerebral cortex slices after the transiently enhanced metabolic activity.

    PubMed

    Sasaki, Toru; Awaji, Takuji; Shimada, Kazuyoshi; Sasaki, Haruyo

    2017-10-01

    Under certain conditions such as hypoxia-reoxygenation, the generation of reactive oxygen species (ROS) increases following hypoxia caused by a decreased oxygen supply. As another hypoxic condition, an excess neural activity status including epileptic seizure induces a decrease in tissue oxygen partial pressure (pO2) caused by enhanced oxygen utilization; however, whether ROS generation increases following the hypoxic status induced by transiently enhanced energy metabolism in brain tissue currently remains unknown. We herein investigated ROS-dependent chemiluminescence in cerebral cortex slices during the restoration of transiently enhanced energy metabolism induced by a high-potassium treatment with tissue pO2 changes and redox balance. ROS generation in the tissue was enhanced after high-potassium-induced hypoxia, but not by the reversed order of the treatment: control-potassium then high-potassium treatment, high-potassium treatment alone, and control-potassium treatment alone. The high-potassium treatment induced a transient decrease in tissue pO2 and a shift in the tissue redox balance towards reduction. The transient shift in the tissue redox balance towards reduction with enhanced metabolic activity and its recovery may correlate with ROS generation. This phenomenon may mimic ROS generation following the hypoxic status induced by transiently enhanced energy metabolism. Copyright © 2017 Elsevier Ireland Ltd and Japan Neuroscience Society. All rights reserved.

  3. 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

  4. The cholesterol-binding protein NPC2 restrains recruitment of stromal macrophage-lineage cells to early-stage lung tumours

    PubMed Central

    Kamata, Tamihiro; Jin, Hong; Giblett, Susan; Patel, Bipin; Patel, Falguni; Foster, Charles; Pritchard, Catrin

    2015-01-01

    The tumour microenvironment is known to play an integral role in facilitating cancer progression at advanced stages, but its function in some pre-cancerous lesions remains elusive. We have used the V600EBRAF-driven mouse lung model that develop premalignant lesions to understand stroma–tumour interactions during pre-cancerous development. In this model, we have found that immature macrophage-lineage cells (IMCs) producing PDGFA, TGFβ and CC chemokines are recruited to the stroma of premalignant lung adenomas through CC chemokine receptor 1 (CCR1)-dependent mechanisms. Stromal IMCs promote proliferation and transcriptional alterations suggestive of epithelial–mesenchymal transition in isolated premalignant lung tumour cells ex vivo, and are required for the maintenance of early-stage lung tumours in vivo. Furthermore, we have found that IMC recruitment to the microenvironment is restrained by the cholesterol-binding protein, Niemann-Pick type C2 (NPC2). Studies on isolated cells ex vivo confirm that NPC2 is secreted from tumour cells and is taken up by IMCs wherein it suppresses secretion of the CCR1 ligand CC chemokine 6 (CCL6), at least in part by facilitating its lysosomal degradation. Together, these findings show that NPC2 secreted by premalignant lung tumours suppresses IMC recruitment to the microenvironment in a paracrine manner, thus identifying a novel target for the development of chemopreventive strategies in lung cancer. PMID:26183450

  5. 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

  6. 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

  7. 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

  8. 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.

  9. Enhanced Ionospheric Plasma Bubble Generation in More Active Atmospheric Intertropical Convergence Zone

    NASA Astrophysics Data System (ADS)

    Yue, X.; Li, G.; Otsuka, Y.; Ning, B.; Abdu, M. A.; Yamamoto, M.; Wan, W.; Liu, L.; Abadi, P.

    2016-12-01

    Using two low latitude steerable backscatter radars in Southeast Asia, the Kototabang (0.2°S, 100.3°E; dip lat 10.4°S) and Sanya (18.4°N, 109.6°E; dip lat 12.8°N) VHF radars, the characteristics of equatorial plasma bubbles (backscatter plumes) during the autumn equinox of 2012-2013 over the two sites separated in longitude by 1000 km were simultaneously investigated. The beam steering measurements reveal frequent occurrences of multiple plumes over both radar sites. We find that the total numbers of nights with plume/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 Intertropical Convergence Zone (ITCZ) is situated around the longitude of Kototabang. Considering that the two stations are separated only by about 9° in longitude where the magnetic declination and the magnetic equator offset from the geographic equator are almost the same, we surmise that the evening pre-reversal enhancement of the eastward electric field (PRE) was unlikely to cause any difference in the bubble generation rate between the two longitudes. The enhanced ionospheric bubble generation at Kototabang longitude could be caused by a higher gravity wave activity associated with the more active ITCZ.

  10. 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.

  11. 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.

  12. 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.

  13. An embryonic myosin converter domain influences Drosophila indirect flight muscle stretch activation, power generation and flight.

    PubMed

    Wang, Qian; Newhard, Christopher S; Ramanath, Seemanti; Sheppard, Debra; Swank, Douglas M

    2014-01-15

    Stretch activation (SA) is critical to the flight ability of insects powered by asynchronous, indirect flight muscles (IFMs). An essential muscle protein component for SA and power generation is myosin. Which structural domains of myosin are significant for setting SA properties and power generation levels is poorly understood. We made use of the transgenic techniques and unique single muscle myosin heavy chain gene of Drosophila to test the influence of the myosin converter domain on IFM SA and power generation. Replacing the endogenous converter with an embryonic version decreased SA tension and the rate of SA tension generation. The alterations in SA properties and myosin kinetics from the converter exchange caused power generation to drop to 10% of control fiber power when the optimal conditions for control fibers - 1% muscle length (ML) amplitude and 150 Hz oscillation frequency - were applied to fibers expressing the embryonic converter (IFI-EC). Optimizing conditions for IFI-EC fiber power production, by doubling ML amplitude and decreasing oscillation frequency by 60%, improved power output to 60% of optimized control fiber power. IFI-EC flies altered their aerodynamic flight characteristics to better match optimal fiber power generation conditions as wing beat frequency decreased and wing stroke amplitude increased. This enabled flight in spite of the drastic changes to fiber mechanical performance.

  14. 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

  15. 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).

  16. Optically active sum-frequency generation as an advanced tool for chiral metallopolymer material

    NASA Astrophysics Data System (ADS)

    Taupier, Grégory; Torres-Werlé, Maria; Boeglin, Alex; Maisse-François, Aline; Achard, Thierry; Bellemin-Laponnaz, Stéphane; Dorkenoo, Kokou Dodzi Honorat

    2017-01-01

    Metallopolymers incorporating metal ions and polytopic ligands offer the advantage to be easily obtained through a self-assembly process in solution but offer great prospects in the development of multifunctional, smart, even self-healing, materials. We have found that chiral enantiopure ligands containing bis(oxazoline) units in combination with Ni(II) salts generate well-defined thin films either by drop casting or by spin-coating and we demonstrate that the condensation process of these chiral metallosupramolecular assemblies can be characterized through optically active sum-frequency generation.

  17. Effects of microbial activity on perfluorinated carboxylic acids (PFCAs) generation during aerobic biotransformation of fluorotelomer alcohols in activated sludge.

    PubMed

    Yu, Xiaolong; Nishimura, Fumitake; Hidaka, Taira

    2017-08-18

    Biotransformation of fluorotelomer alcohols (FTOHs) in wastewater treatment plants (WWTPs) can release toxic intermediates and perfluorinated carboxylic acids (PFCAs) to the aqueous environment. However, little information is known about the role of relevant microbial activity (i.e., autotrophs and/or heterotrophs) in biotransformation of FTOHs. Additionally, the dynamics of microbial community in sludge after exposure to FTOHs remain unclear. In the present research, using domestic and industrial WWTP sludge, we performed lab-scale batch experiments to characterize the FTOHs biodegradation property under aerobic condition. Both heterotrophs and the autotrophs were associated with FTOHs biotransformation. However, the microbial activity influenced PFCAs generation efficiency. Autotrophs based on ammonia oxidation (50mgN/L) resulted in more effective generation of PFCAs than heterotrophs based on glucose (200mgC/L) metabolism. Moreover, autotrophs generated more amounts of short-chain PFCAs (carbon number ≤7) than the heterotrophs. The ammonia monooxygenase (AMO) in ammonia oxidizing microorganisms (AOMs) are suggested as responsible for the enhanced generation of PFCAs during FTOHs biotransformation. In the sludge that had been exposed to poly- and perfluorinated alkyl substances in an industrial WWTP, Chlorobi was the predominant microorganisms (36.9%), followed by Proteobacteria (20.2%), Bacteroidetes (11.1%), Chloroflexi (6.2%), Crenarchaeota (5.6%), Planctomycetes (4.2%), and Acidobacteria (3.5%). In the present research, the dosed 8:2 FTOH (12.1mg/L) and its biotransformation products (intermediates and PFCAs) could force a shift in microbial community composition in the sludge. After 192h, Proteobacteria significantly increased and dominated. These results provide knowledge for comprehending the effects of microbial activity on FTOHs biodegradation and the information about interaction between microbial community and the exposure to FTOHs in activated

  18. 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.

  19. Active torque generation by the actomyosin cell cortex drives left-right symmetry breaking.

    PubMed

    Naganathan, Sundar Ram; Fürthauer, Sebastian; Nishikawa, Masatoshi; Jülicher, Frank; Grill, Stephan W

    2014-12-17

    Many developmental processes break left-right (LR) symmetry with a consistent handedness. LR asymmetry emerges early in development, and in many species the primary determinant of this asymmetry has been linked to the cytoskeleton. However, the nature of the underlying chirally asymmetric cytoskeletal processes has remained elusive. In this study, we combine thin-film active chiral fluid theory with experimental analysis of the C. elegans embryo to show that the actomyosin cortex generates active chiral torques to facilitate chiral symmetry breaking. Active torques drive chiral counter-rotating cortical flow in the zygote, depend on myosin activity, and can be altered through mild changes in Rho signaling. Notably, they also execute the chiral skew event at the 4-cell stage to establish the C. elegans LR body axis. Taken together, our results uncover a novel, large-scale physical activity of the actomyosin cytoskeleton that provides a fundamental mechanism for chiral morphogenesis in development.

  20. 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.

  1. 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.

  2. Metformin Facilitates Amyloid-β Generation by β- and γ-Secretases via Autophagy Activation.

    PubMed

    Son, Sung Min; Shin, Hong-Joon; Byun, Jayoung; Kook, Sun Young; Moon, Minho; Chang, Yu Jin; Mook-Jung, Inhee

    2016-01-01

    The evidence of strong pathological associations between type 2 diabetes and Alzheimer's disease (AD) has increased in recent years. Contrary to suggestions that anti-diabetes drugs may have potential for treating AD, we demonstrate here that the insulin sensitizing anti-diabetes drug metformin (Glucophage®) increased the generation of amyloid-β (Aβ), one of the major pathological hallmarks of AD, by promoting β- and γ-secretase-mediated cleavage of amyloid-β protein precursor (AβPP) in SH-SY5Y cells. In addition, we show that metformin caused autophagosome accumulation in Tg6799 AD model mice. Extremely high γ-secretase activity was also detected in autophagic vacuoles, apparently a novel site of Aβ peptide generation. Together, these data suggest that metformin-induced accumulation of autophagosomes resulted in increased γ-secretase activity and Aβ generation. Additional experiments indicated that metformin increased phosphorylation of AMP-activated protein kinase, which activates autophagy by suppressing mammalian target of rapamycin (mTOR). The suppression of mTOR then induces the abnormal accumulation of autophagosomes. We conclude that metformin, an anti-diabetes drug, may exacerbate AD pathogenesis by promoting amyloidogenic AβPP processing in autophagosomes.

  3. Fabrication Processes to Generate Concentration Gradients in Polymer Solar Cell Active Layers.

    PubMed

    Inaba, Shusei; Vohra, Varun

    2017-05-09

    Polymer solar cells (PSCs) are considered as one of the most promising low-cost alternatives for renewable energy production with devices now reaching power conversion efficiencies (PCEs) above the milestone value of 10%. These enhanced performances were achieved by developing new electron-donor (ED) and electron-acceptor (EA) materials as well as finding the adequate morphologies in either bulk heterojunction or sequentially deposited active layers. In particular, producing adequate vertical concentration gradients with higher concentrations of ED and EA close to the anode and cathode, respectively, results in an improved charge collection and consequently higher photovoltaic parameters such as the fill factor. In this review, we relate processes to generate active layers with ED-EA vertical concentration gradients. After summarizing the formation of such concentration gradients in single layer active layers through processes such as annealing or additives, we will verify that sequential deposition of multilayered active layers can be an efficient approach to remarkably increase the fill factor and PCE of PSCs. In fact, applying this challenging approach to fabricate inverted architecture PSCs has the potential to generate low-cost, high efficiency and stable devices, which may revolutionize worldwide energy demand and/or help develop next generation devices such as semi-transparent photovoltaic windows.

  4. Fabrication Processes to Generate Concentration Gradients in Polymer Solar Cell Active Layers

    PubMed Central

    Inaba, Shusei; Vohra, Varun

    2017-01-01

    Polymer solar cells (PSCs) are considered as one of the most promising low-cost alternatives for renewable energy production with devices now reaching power conversion efficiencies (PCEs) above the milestone value of 10%. These enhanced performances were achieved by developing new electron-donor (ED) and electron-acceptor (EA) materials as well as finding the adequate morphologies in either bulk heterojunction or sequentially deposited active layers. In particular, producing adequate vertical concentration gradients with higher concentrations of ED and EA close to the anode and cathode, respectively, results in an improved charge collection and consequently higher photovoltaic parameters such as the fill factor. In this review, we relate processes to generate active layers with ED–EA vertical concentration gradients. After summarizing the formation of such concentration gradients in single layer active layers through processes such as annealing or additives, we will verify that sequential deposition of multilayered active layers can be an efficient approach to remarkably increase the fill factor and PCE of PSCs. In fact, applying this challenging approach to fabricate inverted architecture PSCs has the potential to generate low-cost, high efficiency and stable devices, which may revolutionize worldwide energy demand and/or help develop next generation devices such as semi-transparent photovoltaic windows. PMID:28772878

  5. 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. Copyright 2010 Elsevier Inc. All rights reserved.

  6. MEG source localization of spatially extended generators of epileptic activity: comparing entropic and hierarchical bayesian approaches.

    PubMed

    Chowdhury, Rasheda Arman; Lina, Jean Marc; Kobayashi, Eliane; Grova, Christophe

    2013-01-01

    Localizing the generators of epileptic activity in the brain using Electro-EncephaloGraphy (EEG) or Magneto-EncephaloGraphy (MEG) signals is of particular interest during the pre-surgical investigation of epilepsy. Epileptic discharges can be detectable from background brain activity, provided they are associated with spatially extended generators. Using realistic simulations of epileptic activity, this study evaluates the ability of distributed source localization methods to accurately estimate the location of the generators and their sensitivity to the spatial extent of such generators when using MEG data. Source localization methods based on two types of realistic models have been investigated: (i) brain activity may be modeled using cortical parcels and (ii) brain activity is assumed to be locally smooth within each parcel. A Data Driven Parcellization (DDP) method was used to segment the cortical surface into non-overlapping parcels and diffusion-based spatial priors were used to model local spatial smoothness within parcels. These models were implemented within the Maximum Entropy on the Mean (MEM) and the Hierarchical Bayesian (HB) source localization frameworks. We proposed new methods in this context and compared them with other standard ones using Monte Carlo simulations of realistic MEG data involving sources of several spatial extents and depths. Detection accuracy of each method was quantified using Receiver Operating Characteristic (ROC) analysis and localization error metrics. Our results showed that methods implemented within the MEM framework were sensitive to all spatial extents of the sources ranging from 3 cm(2) to 30 cm(2), whatever were the number and size of the parcels defining the model. To reach a similar level of accuracy within the HB framework, a model using parcels larger than the size of the sources should be considered.

  7. MEG Source Localization of Spatially Extended Generators of Epileptic Activity: Comparing Entropic and Hierarchical Bayesian Approaches

    PubMed Central

    Chowdhury, Rasheda Arman; Lina, Jean Marc; Kobayashi, Eliane; Grova, Christophe

    2013-01-01

    Localizing the generators of epileptic activity in the brain using Electro-EncephaloGraphy (EEG) or Magneto-EncephaloGraphy (MEG) signals is of particular interest during the pre-surgical investigation of epilepsy. Epileptic discharges can be detectable from background brain activity, provided they are associated with spatially extended generators. Using realistic simulations of epileptic activity, this study evaluates the ability of distributed source localization methods to accurately estimate the location of the generators and their sensitivity to the spatial extent of such generators when using MEG data. Source localization methods based on two types of realistic models have been investigated: (i) brain activity may be modeled using cortical parcels and (ii) brain activity is assumed to be locally smooth within each parcel. A Data Driven Parcellization (DDP) method was used to segment the cortical surface into non-overlapping parcels and diffusion-based spatial priors were used to model local spatial smoothness within parcels. These models were implemented within the Maximum Entropy on the Mean (MEM) and the Hierarchical Bayesian (HB) source localization frameworks. We proposed new methods in this context and compared them with other standard ones using Monte Carlo simulations of realistic MEG data involving sources of several spatial extents and depths. Detection accuracy of each method was quantified using Receiver Operating Characteristic (ROC) analysis and localization error metrics. Our results showed that methods implemented within the MEM framework were sensitive to all spatial extents of the sources ranging from 3 cm2 to 30 cm2, whatever were the number and size of the parcels defining the model. To reach a similar level of accuracy within the HB framework, a model using parcels larger than the size of the sources should be considered. PMID:23418485

  8. Calcium and stretch activation modulate power generation in Drosophila flight muscle.

    PubMed

    Wang, Qian; Zhao, Cuiping; Swank, Douglas M

    2011-11-02

    Many animals regulate power generation for locomotion by varying the number of muscle fibers used for movement. However, insects with asynchronous flight muscles may regulate the power required for flight by varying the calcium concentration ([Ca(2+)]). In vivo myoplasmic calcium levels in Drosophila flight muscle have been found to vary twofold during flight and to correlate with aerodynamic power generation and wing beat frequency. This mechanism can only be possible if [Ca(2+)] also modulates the flight muscle power output and muscle kinetics to match the aerodynamic requirements. We found that the in vitro power produced by skinned Drosophila asynchronous flight muscle fibers increased with increasing [Ca(2+)]. Positive muscle power generation started at pCa = 5.8 and reached its maximum at pCa = 5.25. A twofold variation in [Ca(2+)] over the steepest portion of this curve resulted in a two- to threefold variation in power generation and a 1.2-fold variation in speed, matching the aerodynamic requirements. To determine the mechanism behind the variation in power, we analyzed the tension response to muscle fiber-lengthening steps at varying levels of [Ca(2+)]. Both calcium-activated and stretch-activated tensions increased with increasing [Ca(2+)]. However, calcium tension saturated at slightly lower [Ca(2+)] than stretch-activated tension, such that as [Ca(2+)] increased from pCa = 5.7 to pCa = 5.4 (the range likely used during flight), stretch- and calcium-activated tension contributed 80% and 20%, respectively, to the total tension increase. This suggests that the response of stretch activation to [Ca(2+)] is the main mechanism by which power is varied during flight. Copyright © 2011 Biophysical Society. Published by Elsevier Inc. All rights reserved.

  9. 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.

  10. 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

  11. Systems biology of coagulation initiation: kinetics of thrombin generation in resting and activated human blood.

    PubMed

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

    2010-09-30

    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 (T(i)) of 1 to 2 hours (depending on donor), while activation of platelets with the GPVI-activator convulxin reduces T(i) 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 T(i) 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

  12. 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.

  13. 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.

  14. 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

  15. Generation of Active Protein Phosphatase 2A Is Coupled to Holoenzyme Assembly

    PubMed Central

    Hombauer, Hans; Weismann, David; Mudrak, Ingrid; Stanzel, Claudia; Fellner, Thomas; Lackner, Daniel H; Ogris, Egon

    2007-01-01

    Protein phosphatase 2A (PP2A) is a prime example of the multisubunit architecture of protein serine/threonine phosphatases. Until substrate-specific PP2A holoenzymes assemble, a constitutively active, but nonspecific, catalytic C subunit would constitute a risk to the cell. While it has been assumed that the severe proliferation impairment of yeast lacking the structural PP2A subunit, TPD3, is due to the unrestricted activity of the C subunit, we recently obtained evidence for the existence of the C subunit in a low-activity conformation that requires the RRD/PTPA proteins for the switch into the active conformation. To study whether and how maturation of the C subunit is coupled with holoenzyme assembly, we analyzed PP2A biogenesis in yeast. Here we show that the generation of the catalytically active C subunit depends on the physical and functional interaction between RRD2 and the structural subunit, TPD3. The phenotype of the tpd3Δ strain is therefore caused by impaired, rather than increased, PP2A activity. TPD3/RRD2-dependent C subunit maturation is under the surveillance of the PP2A methylesterase, PPE1, which upon malfunction of PP2A biogenesis, prevents premature generation of the active C subunit and holoenzyme assembly by counteracting the untimely methylation of the C subunit. We propose a novel model of PP2A biogenesis in which a tightly controlled activation cascade protects cells from untargeted activity of the free catalytic PP2A subunit. PMID:17550305

  16. How Activism Features in the Career Lives of Four Generations of Canadian Nurses.

    PubMed

    MacDonnell, Judith A; Buck-McFadyen, Ellen

    2016-11-01

    Recent nursing research using a critical feminist lens challenges the prevailing view of political inertia in nursing. This comparative life history study using a critical feminist lens explores the relevance of activism with four generations of Canadian nurses. Purposeful sampling of Ontario nurses resulted in 40 participants who were diverse in terms of generation, practice setting, and activist practice. Interviews and focus groups were completed with the sample of Ontario registered nurses or undergraduate and graduate nursing students: 8 Generation X, 9 Generation Y (Millennials), 20 Boomers, and 3 Overboomers. Factors such as professional norms and personal and organizational supports shaped contradictory nursing activist identities, practices, and impacts. Gendered norms, organizational dynamics, and the political landscape influenced the meanings nurses attributed to critical incidents and influences that prompted activism inside and outside the workplace, shaping the transformative potential of nursing. Despite its limitations, the study has implications for creating professional and organizational supports for consideration of health politics and policy, and spaces for dialogue to support practice and research aligned with social justice goals.

  17. 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.

  18. 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.

  19. Protrusion force microscopy reveals oscillatory force generation and mechanosensing activity of human macrophage podosomes

    NASA Astrophysics Data System (ADS)

    Labernadie, Anna; Bouissou, Anaïs; Delobelle, Patrick; Balor, Stéphanie; Voituriez, Raphael; Proag, Amsha; Fourquaux, Isabelle; Thibault, Christophe; Vieu, Christophe; Poincloux, Renaud; Charrière, Guillaume M.; Maridonneau-Parini, Isabelle

    2014-11-01

    Podosomes are adhesion structures formed in monocyte-derived cells. They are F-actin-rich columns perpendicular to the substrate surrounded by a ring of integrins. Here, to measure podosome protrusive forces, we designed an innovative experimental setup named protrusion force microscopy (PFM), which consists in measuring by atomic force microscopy the deformation induced by living cells onto a compliant Formvar sheet. By quantifying the heights of protrusions made by podosomes onto Formvar sheets, we estimate that a single podosome generates a protrusion force that increases with the stiffness of the substratum, which is a hallmark of mechanosensing activity. We show that the protrusive force generated at podosomes oscillates with a constant period and requires combined actomyosin contraction and actin polymerization. Finally, we elaborate a model to explain the mechanical and oscillatory activities of podosomes. Thus, PFM shows that podosomes are mechanosensing cell structures exerting a protrusive force.

  20. A novel holding mechanism for next generation active wireless capsule endoscopy.

    PubMed

    Woods, Stephen P; Constandinou, Timothy G

    2015-08-01

    This paper proposes that next generation wireless capsule endoscopy (WCE) technology will feature active mechanical components (i.e. actuated) as opposed to current systems that are predominantly passive (e.g. for imaging purposes). Future systems will integrate microsystems that use micro-actuators to, for example, perform micro-surgery, take tissue samples, deliver medication, etc. In this paper we detail a novel, ultra-compact integrated mechanism for resisting peristalsis and describe how this can be fabricated in Nylon 6 using CNC milling. The holding action is achieved by extending an "anchor" spanning an effective 60.4mm circumference, for a 11.0mm diameter WCE. This function is achieved by a mechanism that occupies only 347.0mm(3) volume, including mechanics and actuator. This shows how exploiting conventional manufacturing processes can result in a radical change in the capabilities of WCE systems and empower the next generation of active devices.

  1. 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.

  2. 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.

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

    PubMed

    Munoz-Organero, Mario; Ruiz-Blazquez, Ramona

    2017-02-08

    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.

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

    DTIC Science & Technology

    2009-03-07

    reducing strain ( Geobacter sulfurreducens ) produced less power than a mixed culture dominated by the same strain. Using a newly developed U-tube MFC, we...Pictures by D. Xing). CBET 0730359, Pi-Logan. Microbial fuel cell architecture Comparison of electrode reduction activities of Geobacter sulfurreducens ...and an enriched consortium in an air-cathode microbial fuel cell. An electricity generating bacterium, Geobacter sulfurreducens PCA, was inoculated

  6. Discovery of BMS-955176, a Second Generation HIV-1 Maturation Inhibitor with Broad Spectrum Antiviral Activity

    PubMed Central

    2016-01-01

    HIV-1 maturation inhibition (MI) has been clinically validated as an approach to the control of HIV-1 infection. However, identifying an MI with both broad polymorphic spectrum coverage and good oral exposure has been challenging. Herein, we describe the design, synthesis, and preclinical characterization of a potent, orally active, second generation HIV-1 MI, BMS-955176 (2), which is currently in Phase IIb clinical trials as part of a combination antiretroviral regimen. PMID:27326328

  7. Discovery of BMS-955176, a Second Generation HIV-1 Maturation Inhibitor with Broad Spectrum Antiviral Activity.

    PubMed

    Regueiro-Ren, Alicia; Liu, Zheng; Chen, Yan; Sin, Ny; Sit, Sing-Yuen; Swidorski, Jacob J; Chen, Jie; Venables, Brian L; Zhu, Juliang; Nowicka-Sans, Beata; Protack, Tricia; Lin, Zeyu; Terry, Brian; Samanta, Himadri; Zhang, Sharon; Li, Zhufang; Beno, Brett R; Huang, Xiaohua S; Rahematpura, Sandhya; Parker, Dawn D; Haskell, Roy; Jenkins, Susan; Santone, Kenneth S; Cockett, Mark I; Krystal, Mark; Meanwell, Nicholas A; Hanumegowda, Umesh; Dicker, Ira B

    2016-06-09

    HIV-1 maturation inhibition (MI) has been clinically validated as an approach to the control of HIV-1 infection. However, identifying an MI with both broad polymorphic spectrum coverage and good oral exposure has been challenging. Herein, we describe the design, synthesis, and preclinical characterization of a potent, orally active, second generation HIV-1 MI, BMS-955176 (2), which is currently in Phase IIb clinical trials as part of a combination antiretroviral regimen.

  8. 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.

  9. 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.

  10. 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.

  11. [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.

  12. 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

  13. 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.

  14. Modulation of Na+/K+ ATPase Activity by Hydrogen Peroxide Generated through Heme in L. amazonensis

    PubMed Central

    Rocco-Machado, Nathália; Cosentino-Gomes, Daniela; Meyer-Fernandes, José Roberto

    2015-01-01

    Leishmania amazonensis is a protozoan parasite that occurs in many areas of Brazil and causes skin lesions. Using this parasite, our group showed the activation of Na+/K+ ATPase through a signaling cascade that involves the presence of heme and protein kinase C (PKC) activity. Heme is an important biomolecule that has pro-oxidant activity and signaling capacity. Reactive oxygen species (ROS) can act as second messengers, which are required in various signaling cascades. Our goal in this work is to investigate the role of hydrogen peroxide (H2O2) generated in the presence of heme in the Na+/K+ ATPase activity of L. amazonensis. Our results show that increasing concentrations of heme stimulates the production of H2O2 in a dose-dependent manner until a concentration of 2.5 μM heme. To confirm that the effect of heme on the Na+/K+ ATPase is through the generation of H2O2, we measured enzyme activity using increasing concentrations of H2O2 and, as expected, the activity increased in a dose-dependent manner until a concentration of 0.1 μM H2O2. To investigate the role of PKC in this signaling pathway, we observed the production of H2O2 in the presence of its activator phorbol 12-myristate 13-acetate (PMA) and its inhibitor calphostin C. Both showed no effect on the generation of H2O2. Furthermore, we found that PKC activity is increased in the presence of H2O2, and that in the presence of calphostin C, H2O2 is unable to activate the Na+/K+ ATPase. 100 μM of Mito-TEMPO was capable of abolishing the stimulatory effect of heme on Na+/K+ ATPase activity, indicating that mitochondria might be the source of the hydrogen peroxide production induced by heme. The modulation of L. amazonensis Na+/K+ ATPase by H2O2 opens new possibilities for understanding the signaling pathways of this parasite. PMID:26070143

  15. 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

  16. 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

  17. 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

  18. Chloride channels activated by swell can regulate the NADPH oxidase generated membrane depolarisation in activated human neutrophils

    SciTech Connect

    Ahluwalia, Jatinder

    2008-01-11

    Chloride channels activated by swell have important functions in many physiological processes. The phagocyte NADPH oxidase is essential for host defence and it generates superoxide by transferring electrons from the donor NADPH to the acceptor O{sub 2}. This electron current, induces a depolarisation of the plasma membrane. In this study, I report that chloride channels activated by swell can counteract the depolarisation induced by the NADPH oxidase. When a chloride conductance was activated by swelling, its inhibition by either 50 {mu}M NPPB or removing external chloride, depolarised the plasma membrane potential to +26 mV {+-} 3.1 (n = 4) and +40 {+-} 1 mV (n = 4), respectively. These channels were partially inhibited by the NADPH oxidase inhibitor AEBSF (1 mM) and potently inhibited by ZnCl{sub 2} (3 mM). These currents were not activated by a phosphorylation step and elevations in intracellular calcium did not appear to activate chloride currents similar to those activated by swell.

  19. Microbicidal Activity of Vascular Peroxidase 1 in Human Plasma via Generation of Hypochlorous Acid

    PubMed Central

    Li, Hong; Cao, Zehong; Moore, D. Ray; Jackson, Patricia L.; Barnes, Stephen; Lambeth, J. David; Thannickal, Victor J.

    2012-01-01

    Members of the heme peroxidase family play an important role in host defense. Myeloperoxidase (MPO) is expressed in phagocytes and is the only animal heme peroxidase previously reported to be capable of using chloride ion as a substrate to form the highly microbicidal species hypochlorous acid (HOCl) at neutral pH. Despite the potent bacterial killing activity of HOCl, individuals who fail to express MPO typically show only a modest increase in some fungal infections. This may point to the existence of redundant host defense mechanisms. Vascular peroxidase 1 (VPO1) is newly discovered member of the heme peroxidase family. VPO1 is expressed in cells of the cardiovascular system and is secreted into the bloodstream. In the present study, we investigate whether VPO1 is capable of generating HOCl and its role in host defense. Like MPO, VPO1 in the presence of H2O2 and chloride generates HOCl. VPO1-dependent HOCl generation was demonstrated by chlorination of taurine and tyrosine using mass spectrometry. In addition, the VPO1/H2O2/Cl− system can cause the chlorination of monochlorodimedone and the oxidation of 5-thio-2-nitrobenzoic acid. Purified VPO1 and VPO1 in plasma mediate bacterial killing that is dependent on chloride and H2O2; killing is inhibited by peroxidase inhibitors and by the H2O2 scavenger catalase. In the presence of erythrocytes, bacterial killing by VPO1 is slightly reduced. Thus, VPO1, in addition to MPO, is the second member of the heme peroxidase family capable of generating HOCl under physiological conditions. VPO1 is likely to participate in host defense, with bactericidal activity mediated through the generation of HOCl. PMID:22526679

  20. Spindle oscillations during asymmetric cell division require a threshold number of active cortical force generators.

    PubMed

    Pecreaux, Jacques; Röper, Jens-Christian; Kruse, Karsten; Jülicher, Frank; Hyman, Anthony A; Grill, Stephan W; Howard, Jonathon

    2006-11-07

    Asymmetric division of the C. elegans zygote is due to the posterior-directed movement of the mitotic spindle during metaphase and anaphase. During this movement along the anterior-posterior axis, the spindle oscillates transversely. These motions are thought to be driven by a force-generating complex-possibly containing the motor protein cytoplasmic dynein-that is located at the cell cortex and pulls on microtubules growing out from the spindle poles. A theoretical analysis indicates that the oscillations might arise from mechanical coordination of the force-generating motors, and this coordination is mediated by the load dependence of the motors' detachment from the microtubules. The model predicts that the motor activity must exceed a threshold for oscillations to occur. We have tested the existence of a threshold by using RNA interference to gradually reduce the levels of dynein light intermediate chain as well as GPR-1 and GPR-2 that are involved in the G protein-mediated regulation of the force generators. We found an abrupt cessation of oscillations as expected if the motor activity dropped below a threshold. Furthermore, we can account for the complex choreography of the mitotic spindle-the precise temporal coordination of the buildup and die-down of the transverse oscillations with the posterior displacement-by a gradual increase in the processivity of a single type of motor machinery during metaphase and anaphase. The agreement between our results and modeling suggests that the force generators themselves have the intrinsic capability of generating oscillations when opposing forces exceed a threshold.

  1. 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

  2. The role of cellular coupling in the spontaneous generation of electrical activity in uterine tissue.

    PubMed

    Xu, Jinshan; Menon, Shakti N; Singh, Rajeev; Garnier, Nicolas B; Sinha, Sitabhra; Pumir, Alain

    2015-01-01

    The spontaneous emergence of contraction-inducing electrical activity in the uterus at the beginning of labor remains poorly understood, partly due to the seemingly contradictory observation that isolated uterine cells are not spontaneously active. It is known, however, that the expression of gap junctions increases dramatically in the approach to parturition, by more than one order of magnitude, which results in a significant increase in inter-cellular electrical coupling. In this paper, we build upon previous studies of the activity of electrically excitable smooth muscle cells (myocytes) and investigate the mechanism through which the coupling of these cells to electrically passive cells results in the generation of spontaneous activity in the uterus. Using a recently developed, realistic model of uterine muscle cell dynamics, we investigate a system consisting of a myocyte coupled to passive cells. We then extend our analysis to a simple two-dimensional lattice model of the tissue, with each myocyte being coupled to its neighbors, as well as to a random number of passive cells. We observe that different dynamical regimes can be observed over a range of gap junction conductances: at low coupling strength, corresponding to values measured long before delivery, the activity is confined to cell clusters, while the activity for high coupling, compatible with values measured shortly before delivery, may spread across the entire tissue. Additionally, we find that the system supports the spontaneous generation of spiral wave activity. Our results are both qualitatively and quantitatively consistent with observations from in vitro experiments. In particular, we demonstrate that the increase in inter-cellular electrical coupling observed experimentally strongly facilitates the appearance of spontaneous action potentials that may eventually lead to parturition.

  3. Role of mucosa in generating spontaneous activity in the guinea pig seminal vesicle.

    PubMed

    Takeya, Mitsue; Hashitani, Hikaru; Hayashi, Tokumasa; Higashi, Ryuhei; Nakamura, Kei-Ichiro; Takano, Makoto

    2017-07-15

    The mucosa may have neuron-like functions as urinary bladder mucosa releases bioactive substances that modulate sensory nerve activity as well as detrusor muscle contractility. However, such mucosal function in other visceral organs remains to be established. The role of mucosa in generating spontaneous contractions in seminal vesicles (SVs), a paired organ in the male reproductive tract, was investigated. The intact mucosa is essential for the generation of spontaneous phasic contractions of SV smooth muscle arising from electrical slow waves and corresponding increases in intracellular Ca(2+) . These spontaneous events primarily depend on Ca(2+) handling by sarco-endoplasmic reticulum Ca(2+) stores. A population of mucosal cells developed spontaneous rises in intracellular Ca(2+) relying on sarco-endoplasmic reticulum Ca(2+) handling. The spontaneously active cells in the SV mucosa appear to drive spontaneous activity in smooth muscle either by sending depolarizing signals and/or by releasing humoral substances. The role of the mucosa in generating the spontaneous activity of guinea-pig seminal vesicle (SV) was explored. Changes in contractility, membrane potential and intracellular Ca(2+) dynamics of SV smooth muscle cells (SMCs) were recorded using isometric tension recording, intracellular microelectrode recording and epi-fluorescence Ca(2+) imaging, respectively. Mucosa-intact but not mucosa-denuded SV preparations generated TTX- (1 μm) resistant spontaneous phasic contractions that were abolished by nifedipine (3 μm). Consistently, SMCs developed mucosa-dependent slow waves (SWs) that triggered action potentials and corresponding Ca(2+) flashes. Nifedipine (10 μm) abolished the action potentials and spontaneous contractions, while suppressing the SWs and Ca(2+) flashes. Both the residual SWs and spontaneous Ca(2+) transients were abolished by cyclopiazonic acid (CPA, 10 μm), a sarco-endoplasmic reticulum Ca(2+) -ATPase (SERCA) inhibitor. DIDS (300

  4. Superoxide radical generation and Mn- and Cu-Zn superoxide dismutases activities in human leukemic cells.

    PubMed

    Kato, Masahiko; Minakami, Hisanori; Kuroiwa, Minoru; Kobayashi, Yasuyuki; Oshima, Shigeru; Kozawa, Kunihisa; Morikawa, Akihiro; Kimura, Hirokazu

    2003-03-01

    Mn- and Cu-Zn superoxide dismutase (SOD) activities and generation of superoxide radicals (O(2) (-)) were assessed in leukemic cells from 10 patients with acute myeloid or monocytic leukemia (AML) and 10 patients with acute lymphoblastic leukemia (ALL), using a sensitive, specific chemiluminescence method. Leukemic cells were classified according to the French-American-British classification. M4 AML cells from two patients produced some O(2) (-) upon stimulation with opsonized zymosan (OZ), phorbol myristate acetate (PMA) or N-formyl-methionyl-leucyl-phenylalanine (FMLP), but less than normal granulocytes or monocytes. M5b AML cells from one patient produced as much O(2) (-) in response to these stimulants as normal monocytes. No O(2) (-) generation was induced in other types of leukemic cells. Total SOD activity in AML cells was significantly greater in normal granulocytes, but was only half of the activity in ALL cells. Mn-SOD in AML cells was very low or undetectable. These results suggest that except in M5b cells, decreased O(2) (-) production may contribute to susceptibility to infections in AML patients. Decreased Mn-SOD activity in AML cells may predispose them to oxidative stress. Copyright 2002 John Wiley & Sons, Ltd.

  5. 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.

  6. Engineering streptokinase for generation of active site-labeled plasminogen analogs*

    PubMed Central

    Laha, Malabika; Panizzi, Peter; Nahrendorf, Matthias; Bock, Paul E.

    2011-01-01

    We previously demonstrated that streptokinase (SK) can be used to generate active site-labeled fluorescent analogs of plasminogen (Pg) by virtue of its non-proteolytic activation of the zymogen. The method is versatile and allows for stoichiometric and active site-specific incorporation of any one of many molecular probes. The limitation of the labeling approach is that it is both time-consuming and low yield. Here we demonstrate an improved method for the preparation of labeled Pg analogs by the use of an engineered SK mutant fusion protein with both COOH- and NH2-terminal His6-tags. The NH2-terminal tag is followed by a tobacco etch virus proteinase cleavage site to ensure that the SK Ile1 residue, essential for conformational activation of Pg, is preserved. The SK COOH-terminal Lys414 residue and residues Arg253-Leu260 in the SK β-domain were deleted to prevent cleavage by plasmin (Pm), and to disable Pg substrate binding to the SK·Pg*/Pm catalytic complexes, respectively. Near-elimination of Pm generation with the SKΔ(R253-L260)ΔK414-His6 mutant increased the yield of labeled Pg 2.6-fold and reduced the time required >2-fold. The versatility of the labeling method was extended to the application of Pg labeled with a near-infrared probe to quantitate Pg receptors on immune cells by flow cytometry. PMID:21570944

  7. 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.

  8. 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.

  9. 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

  10. 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.

  11. Distinguishing How from Why the Mind Wanders: A Process-Occurrence Framework for Self-Generated Mental Activity

    ERIC Educational Resources Information Center

    Smallwood, Jonathan

    2013-01-01

    Cognition can unfold with little regard to the events taking place in the environment, and such self-generated mental activity poses a specific set of challenges for its scientific analysis in both cognitive science and neuroscience. One problem is that the spontaneous onset of self-generated mental activity makes it hard to distinguish the events…

  12. Silver-ion-mediated reactive oxygen species generation affecting bactericidal activity.

    PubMed

    Park, Hee-Jin; Kim, Jee Yeon; Kim, Jaeeun; Lee, Joon-Hee; Hahn, Ji-Sook; Gu, Man Bock; Yoon, Jeyong

    2009-03-01

    Silver ions have been widely used as disinfectants that inhibit bacterial growth by inhibiting the essential enzymatic functions of the microorganism via interaction with the thiol-group of l-cysteine. However, silver-ion-mediated perturbation of the bacterial respiratory chain has raised the possibility of reactive oxygen species (ROS) generation. We used bacterial reporter strains specifically responding to superoxide radicals and found that silver-ion-mediated ROS-generation affected bactericidal activity. Almost half the log reduction in Escherichia coli and Staphylococcus aureus populations (model strains for gram negative and positive bacteria, respectively) caused by silver-ion disinfection was attributed to ROS-mediated bactericidal activity. The major form of ROS generated was the superoxide-radical; H(2)O(2) was not induced. Furthermore, silver ions strongly enhanced paraquat-induced oxidative stress, indicating close correlation and synergism between the conventional and ROS-mediated silver toxicity. Our results suggest that further studies in silver-based disinfection systems should consider the oxygen concentration and ROS reaction.

  13. 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

  14. 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

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

    PubMed Central

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

    2013-01-01

    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. PMID:24303193

  16. 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.

  17. Sublethal Caspase Activation Promotes Generation of Cardiomyocytes from Embryonic Stem Cells

    PubMed Central

    Österholm, Cecilia; Wang, Heng; Beltrán-Rodríguez, Antonio; Varas-Godoy, Manuel; Månsson-Broberg, Agneta; Uhlén, Per; Simon, András; Grinnemo, Karl-Henrik

    2015-01-01

    Generation of new cardiomyocytes is critical for cardiac repair following myocardial injury, but which kind of stimuli is most important for cardiomyocyte regeneration is still unclear. Here we explore if apoptotic stimuli, manifested through caspase activation, influences cardiac progenitor up-regulation and cardiomyocyte differentiation. Using mouse embryonic stem cells as a cellular model, we show that sublethal activation of caspases increases the yield of cardiomyocytes while concurrently promoting the proliferation and differentiation of c-Kit+/α-actininlow cardiac progenitor cells. A broad-spectrum caspase inhibitor blocked these effects. In addition, the caspase inhibitor reversed the mRNA expression of genes expressed in cardiomyocytes and their precursors. Our study demonstrates that sublethal caspase-activation has an important role in cardiomyocyte differentiation and may have significant implications for promoting cardiac regeneration after myocardial injury involving exogenous or endogenous cell sources. PMID:25763592

  18. Comparison and Analysis of Inter-Subject Variability of Simulated Magnetic Activity Generated from Gastric Electrical Activity

    PubMed Central

    Komuro, Rie; Cheng, Leo K.; Pullan, Andrew J.

    2014-01-01

    Electrogastrograms (EGGs) produced from gastric electrical activity (GEA) are used as a non-invasive method to aid in the assessment of a subject’s gastric condition. It has been documented that recordings of the magnetic activity generated from GEA are more reliable. Typically, with magnetic measurements of GEA, only activity perpendicular to the body is recorded. Also, external anatomical landmarks are used to position the magnetic recording devices, SQUIDs, (Superconducting Quantum Interference Devices) over the stomach with no allowance made for body habitus. In the work presented here, GEA and its corresponding magnetic activity are simulated. Using these data, we investigate the effects of using a standard SQUID location as well as a customized SQUID position and the contribution the magnetic component perpendicular to the body makes to the magnetic field. We also explore the effects of the stomach wall thickness on the resultant magnetic fields. The simulated results show that the thicker the wall, the larger the magnitude of the magnetic field holding the same signal patterns. We conclude that most of the magnetic activity arising from GEA occurs in a plane parallel to the anterior body. We also conclude that using a standard SQUID position can be suboptimal. PMID:18330701

  19. Comparison and analysis of inter-subject variability of simulated magnetic activity generated from gastric electrical activity.

    PubMed

    Komuro, Rié; Cheng, Leo K; Pullan, Andrew J

    2008-06-01

    Electrogastrograms (EGGs) produced from gastric electrical activity (GEA) are used as a non-invasive method to aid in the assessment of a subject's gastric condition. It has been documented that recordings of the magnetic activity generated from GEA are more reliable. Typically, with magnetic measurements of GEA, only activity perpendicular to the body is recorded. Also, external anatomical landmarks are used to position the magnetic recording devices, SQUIDs, (Superconducting Quantum Interference Devices) over the stomach with no allowance made for body habitus. In the work presented here, GEA and its corresponding magnetic activity are simulated. Using these data, we investigate the effects of using a standard SQUID location as well as a customized SQUID position and the contribution the magnetic component perpendicular to the body makes to the magnetic field. We also explore the effects of the stomach wall thickness on the resultant magnetic fields. The simulated results show that the thicker the wall, the larger the magnitude of the magnetic field holding the same signal patterns. We conclude that most of the magnetic activity arising from GEA occurs in a plane parallel to the anterior body. We also conclude that using a standard SQUID position can be suboptimal.

  20. Structure of human nSMase2 reveals an interdomain allosteric activation mechanism for ceramide generation.

    PubMed

    Airola, Michael V; Shanbhogue, Prajna; Shamseddine, Achraf A; Guja, Kip E; Senkal, Can E; Maini, Rohan; Bartke, Nana; Wu, Bill X; Obeid, Lina M; Garcia-Diaz, Miguel; Hannun, Yusuf A

    2017-07-11

    Neutral sphingomyelinase 2 (nSMase2, product of the SMPD3 gene) is a key enzyme for ceramide generation that is involved in regulating cellular stress responses and exosome-mediated intercellular communication. nSMase2 is activated by diverse stimuli, including the anionic phospholipid phosphatidylserine. Phosphatidylserine binds to an integral-membrane N-terminal domain (NTD); however, how the NTD activates the C-terminal catalytic domain is unclear. Here, we identify the complete catalytic domain of nSMase2, which was misannotated because of a large insertion. We find the soluble catalytic domain interacts directly with the membrane-associated NTD, which serves as both a membrane anchor and an allosteric activator. The juxtamembrane region, which links the NTD and the catalytic domain, is necessary and sufficient for activation. Furthermore, we provide a mechanistic basis for this phenomenon using the crystal structure of the human nSMase2 catalytic domain determined at 1.85-Å resolution. The structure reveals a DNase-I-type fold with a hydrophobic track leading to the active site that is blocked by an evolutionarily conserved motif which we term the "DK switch." Structural analysis of nSMase2 and the extended N-SMase family shows that the DK switch can adopt different conformations to reposition a universally conserved Asp (D) residue involved in catalysis. Mutation of this Asp residue in nSMase2 disrupts catalysis, allosteric activation, stimulation by phosphatidylserine, and pharmacological inhibition by the lipid-competitive inhibitor GW4869. Taken together, these results demonstrate that the DK switch regulates ceramide generation by nSMase2 and is governed by an allosteric interdomain interaction at the membrane interface.

  1. Single cell active force generation under dynamic loading - Part I: AFM experiments.

    PubMed

    Weafer, P P; Reynolds, N H; Jarvis, S P; McGarry, J P

    2015-11-01

    A novel series of experiments are performed on single cells using a bespoke AFM system where the response of cells to dynamic loading at physiologically relevant frequencies is uncovered. Measured forces for the untreated cells are dramatically different to cytochalasin-D (cyto-D) treated cells, indicating that the contractile actin cytoskeleton plays a critical role in the response of cells to dynamic loading. Following a change in applied strain magnitude, while maintaining a constant applied strain rate, the compression force for contractile cells recovers to 88.9±7.8% of the steady state force. In contrast, cyto-D cell compression forces recover to only 38.0±6.7% of the steady state force. Additionally, untreated cells exhibit strongly negative (pulling) forces during unloading half-cycles when the probe is retracted. In comparison, negligible pulling forces are measured for cyto-D cells during probe retraction. The current study demonstrates that active contractile forces, generated by actin-myosin cross-bridge cycling, dominate the response of single cells to dynamic loading. Such active force generation is shown to be independent of applied strain magnitude. Passive forces generated by the applied deformation are shown to be of secondary importance, exhibiting a high dependence on applied strain magnitude, in contrast to the active forces in untreated cells. A novel series of experiments are performed on single cells using a bespoke AFM system where the response of cells to dynamic loading at physiologically relevant frequencies is uncovered. Contractile cells, which contain the active force generation machinery of the actin cytoskeleton, are shown to be insensitive to applied strain magnitude, exhibiting high resistance to dynamic compression and stretching. Such trends are not observed for cells in which the actin cytoskeleton has been chemically disrupted. These biomechanical insights have not been previously reported. This detailed characterisation of

  2. 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.

  3. 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.

  4. Mechanical and free living comparisons of four generations of the Actigraph activity monitor

    PubMed Central

    2012-01-01

    Background More studies include multiple generations of the Actigraph activity monitor. So far no studies have compared the output including the newest generation and investigated the impact on the output of the activity monitor when enabling the low frequency extension (LFE) option. The aims were to study the responses of four generations (AM7164, GT1M, GT3X and GT3X+) of the Actigraph activity monitor in a mechanical setup and a free living environment with and without enabling the LFE option. Methods The monitors were oscillated in a mechanical setup using two radii in the frequency range 0.25-3.0 Hz. Following the mechanical study a convenience sample (N = 20) wore three monitors (one AM7164 and two GT3X) for 24 hours. Results The AM7164 differed from the newer generations across frequencies (p < 0.05) in the mechanical setup. The AM7164 produced a higher output at the lower and at the highest intensities, whereas the output was lower at the middle intensities in the mid-range compared to the newer generations. The LFE option decreased the differences at the lower frequencies, but increased differences at the higher. In free living, the mean physical activity level (PA) of the GT3X was 18 counts per minute (CPM) (8%) lower compared to the AM7164 (p < 0.001). Time spent in sedentary intensity was 26.6 minutes (95% CI 15.6 to 35.3) higher when assessed by the GT3X compared to the AM7164 (p < 0.001). Time spend in light and vigorous PA were 23.3 minutes (95% CI 31.8 to 14.8) and 11.7 minutes (95% CI 2.8 to 0.7) lower when assessed by the GT3X compared to the AM7164 (p < 0.05). When enabling the LFE the differences in the sedentary and light PA intensity (<333 counts*10 sec-1) were attenuated (p > 0.05 for differences between generations) thus attenuated the difference in mean PA (p > 0.05) when the LFE option was enabled. However, it did not attenuate the difference in time spend in vigorous PA and it introduced a difference

  5. 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

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

    PubMed

    Garden, Derek L F; Rinaldi, Arianna; Nolan, Matthew F

    2017-02-15

    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. 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 calcium-activated

  7. 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.

  8. 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.

  9. Generation of high-frequency electric field activity by turbulence in the Earth's magnetotail

    NASA Astrophysics Data System (ADS)

    Stawarz, J. E.; Ergun, R. E.; Goodrich, K. A.

    2015-03-01

    Bursty bulk flow (BBF) events, frequently observed in the magnetotail, carry significant energy and mass from the tail region at distances that are often greater than 20 RE into the near-Earth plasma sheet at ˜10 RE where the flow is slowed and/or diverted. This region at ˜10 RE is referred to as the BBF braking region. A number of possible channels are available for the transfer or dissipation of energy in BBF events including adiabatic heating of particles, the propagation of Alfvén waves out of the BBF braking region and into the auroral region, diverted flow out of the braking region, and energy dissipation within the braking region itself. This study investigates the generation of intense high-frequency electric field activity observed within the braking region. When present, these intense electric fields have power above the ion cyclotron frequency and almost always contain nonlinear structures such as electron phase space holes and double layers, which are often associated with field-aligned currents. A hypothesis in which the observed high-frequency electric field activity is generated by field-aligned currents resulting from turbulence in the BBF braking region is considered. Although linear Alfvén waves can generate field-aligned currents, based on theoretical calculations, the required currents are likely not the result of linear waves. Observations from the Time History of Events and Macroscale Interactions during Substorms satellites support the picture of a turbulent plasma leading to the generation of nonlinear kinetic structures. This work provides a possible mechanism for energy dissipation in turbulent plasmas.

  10. [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.

  11. Intracellular reactive oxygen species in monocytes generated by photosensitive chromophores activated with blue light.

    PubMed

    Bouillaguet, Serge; Owen, Brandi; Wataha, John C; Campo, Marino A; Lange, Norbert; Schrenzel, Jacques

    2008-08-01

    Disinfection of the tooth pulp-canal system is imperative to successful endodontic therapy. Yet, studies suggest that 30-50% of current endodontic treatments fail from residual bacterial infection. Photodynamic therapy using red-light chromophores (630 nm) to induce antimicrobial death mediated by generated reactive oxygen species (ROS) has been reported, but red-light also may thermally damage resident tissues. In the current study, we tested the hypothesis that several blue light chromophores (380-500 nm) generate intracellular reactive oxygen species but are not cytotoxic to mammalian cells. THP1 monocytes were exposed to 10 microM of four chromophores (chlorin e6, pheophorbide-a, pheophorbide-a-PLL, and riboflavin) for 30 min before activation with blue light (27J/cm(2), 60s). After activation, intracellular ROS were measured using a dihydrofluorescein diacetate technique, and cytotoxicity was determined by measuring mitochondrial activity with the MTT method. All photosensitizers produced intracellular ROS levels that were dependent on both the presence of the photosensitizer and blue light exposure. Riboflavin and pheophorbide-a-PLL produced the highest levels of ROS. Photosensitizers except riboflavin exhibited cytotoxicity above 10 microM, and all except pheophorbide-a-PLL were more cytotoxic after blue light irradiation. The current study demonstrated the possible utility of blue light chromophores as producers of ROS that would be useful for endodontic disinfection.

  12. Chemotherapeutic agents subvert tumor immunity by generating agonists of platelet-activating factor

    PubMed Central

    Sahu, Ravi P.; Ocana, Jesus A.; Harrison, Kathleen A.; Ferracini, Matheus; Touloukian, Christopher E.; Al-Hassani, Mohammed; Sun, Louis; Loesch, Mathew; Murphy, Robert C.; Althouse, Sandra K.; Perkins, Susan M.; Speicher, Paul J.; Tyler, Douglas S.; Konger, Raymond L.; Travers, Jeffrey B.

    2014-01-01

    Oxidative stress suppresses host immunity by generating oxidized lipid agonists of the platelet-activating factor receptor (PAF-R). Because many classical chemotherapeutic drugs induce reactive oxygen species (ROS), we investigated whether these drugs might subvert host immunity by activating PAF-R. Here we show that PAF-R agonists are produced in melanoma cells by chemotherapy that is administered in vitro, in vivo or in human subjects. Structural characterization of the PAF-R agonists induced revealed multiple oxidized glycerophosphocholines that are generated non-enzymatically. In a murine model of melanoma, chemotherapeutic administration could augment tumor growth by a PAF-R-dependent process that could be blocked by treatment with antioxidants or cyclooxygenase-2 inhibitors or by depletion of regulatory T cells. Our findings reveal how PAF-R agonists induced by chemotherapy treatment can promote treatment failure. Further, they offer new insights into how to improve the efficacy of chemotherapy by blocking its heretofore unknown impact on PAF-R activation. PMID:25304264

  13. Theta-Modulated Gamma-Band Synchronization Among Activated Regions During a Verb Generation Task

    PubMed Central

    Doesburg, Sam M.; Vinette, Sarah A.; Cheung, Michael J.; Pang, Elizabeth W.

    2012-01-01

    Expressive language is complex and involves processing within a distributed network of cortical regions. Functional MRI and magnetoencephalography (MEG) have identified brain areas critical for expressive language, but how these regions communicate across the network remains poorly understood. It is thought that synchronization of oscillations between neural populations, particularly at a gamma rate (>30 Hz), underlies functional integration within cortical networks. Modulation of gamma rhythms by theta-band oscillations (4–8 Hz) has been proposed as a mechanism for the integration of local cell coalitions into large-scale networks underlying cognition and perception. The present study tested the hypothesis that these oscillatory mechanisms of functional integration were present within the expressive language network. We recorded MEG while subjects performed a covert verb generation task. We localized activated cortical regions using beamformer analysis, calculated inter-regional phase locking between activated areas, and measured modulation of inter-regional gamma synchronization by theta phase. The results show task-dependent gamma-band synchronization among regions activated during the performance of the verb generation task, and we provide evidence that these transient and periodic instances of high-frequency connectivity were modulated by the phase of cortical theta oscillations. These findings suggest that oscillatory synchronization and cross-frequency interactions are mechanisms for functional integration among distributed brain areas supporting expressive language processing. PMID:22707946

  14. 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.

  15. Mapping Active Stream Lengths as a Tool for Understanding Spatial Variations in Runoff Generation

    NASA Astrophysics Data System (ADS)

    Erwin, E. G.; Gannon, J. P.; Zimmer, M. A.

    2016-12-01

    Recent studies have shown temporary stream channels respond in complex ways to precipitation. By investigating how stream networks expand and recede throughout rain events, we may further develop our understanding of runoff generation. This study focused on mapping the expansion and contraction of the stream network in two headwater catchments characterized by differing soil depths and slopes, located in North Carolina, USA. The first is a 43 ha catchment located in the Southern Appalachian region, characterized by incised, steep slopes and soils of varying thickness. The second is a 3.3 ha catchment located in the Piedmont region, characterized as low relief with deep, highly weathered soils. Over a variety of flow conditions, surveys of the entire stream network were conducted at 10 m intervals to determine presence or absence of surface water. These surveys revealed several reaches within the networks that were intermittent, with perennial flow upstream and downstream. Furthermore, in some tributaries, the active stream head moved up the channel in response to precipitation and at others it remained anchored in place. Moreover, when repeat surveys were performed during the same storm, hysteresis was observed in active stream length variations: stream length was not the same on the rising limb and falling limb of the hydrograph. These observations suggest there are different geomorphological controls or runoff generation processes occurring spatially throughout these catchments. Observations of wide spatial and temporal variability of active stream length over a variety of flow conditions suggest runoff dynamics, generation mechanisms, and contributing flowpath depths producing streamflow may be highly variable and not easily predicted from streamflow observations at a fixed point. Finally, the observation of similar patterns in differing geomorphic regions suggests these processes extend beyond unique site characterizations.

  16. 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.

  17. Antiplatelet Aggregation Activity of Walnut Hull Extract via Suppression of Reactive Oxygen Species Generation and Caspase Activation.

    PubMed

    Meshkini, Azadeh; Tahmasbi, Masoumeh

    2017-06-01

    Walnut hull (wal hull) is an agricultural by-product that is widely used in traditional medicine for alleviating pain and treating skin diseases, however, recently it has gained much attention in modern pharmacology due to its antioxidant properties. The current study was aimed to determine the total phenolic, flavonoid, and tannin content of Persian wal hull extract and evaluate its biological effects on platelet function. Experimental data showed that acetone extract of wal hulls has a high content of polyphenolic compounds and antioxidant properties. The analytical study of crude extract by gas chromatography-mass spectrometry demonstrated different types of high- and low-molecular-weight compounds that are basically and biologically important. Moreover, an in vitro study revealed that wal hull extract at a concentration of 50 μg/mL inhibited thrombin-induced platelet aggregation and protein secretion by 50%, without any cytotoxic effects on platelets. The examined extract suppressed reactive oxygen species generation and also caspase activation in thrombin-stimulated platelets. Identically, N-acetylcysteine inhibited the increase of reactive oxygen species level induced by thrombin in platelets, and supported a link between cellular redox status and caspase activation in activated platelets. Presumably, the antiplatelet activity of wal hull extract is related to its polyphenolic compounds and their antioxidant properties. Therefore, wal hulls can be considered as a candidate for thrombotic disorders. Copyright © 2017. Published by Elsevier B.V.

  18. 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.

  19. 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

  20. A Survey of Combat Models for Use in CARF (Combat Active Replacement Factor) Value Generation

    DTIC Science & Technology

    1985-03-01

    requirements for the degree of MASTER OF SCIENCE IN CPERATIONS RESEARCH from the NAVAL POSTGRADUATE SCHOOL darch 1985 Author: tv Approved by :... - n Z ...GENERATION OF A WARTIME ACTIVE REPLACEMENT FACTOR (WARF) 65 A. THE PREPROCESSER ................. 66 B. THE COMBAT SAMPLE GEERATOR (COSAGE) MODEL 66 C. THE...en -T,, , -T T L T 17, V 7777% T -T T K 97 7 Ř 7 T W 71 T 7 - Z W.; ., . 7. , II. THE CARF GENERA7ION SYSTEM TODAY The previous and current CARF

  1. 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.

  2. 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.

  3. Wind-Driven Coastal Generation of Annual Mesoscale Eddy Activity in the California Current

    DTIC Science & Technology

    1993-02-01

    DATES COVERED 12/93 FINAL 10/ 1 /90-11/30/91 4. TintL AND SUBTITLE S. FUNDING NUMBERS Wind-driven Coastal Generation of Annual Mesoscale Eddy Activity in...winds. The other is a I 1 /2-layer primitive equation (PE) model with the ability to represent the coastal response to winds but not baroclinic...California Current ALEJANDRO PARES-SIERRA* WARREN B. WHITE " AD C_-K TA:* ’Cenifo de InrwoitCenriai£ ivnp S9,PCYw’ . 1 ,r Lnu~naaj (.Msnucnpt tecei~ed 𔄀 S c

  4. 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.

  5. 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

  6. 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.

  7. 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

    2017-05-01

    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. © 2017 Wiley Periodicals, Inc.

  8. 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.

  9. Active torque generation by the actomyosin cell cortex drives left–right symmetry breaking

    PubMed Central

    Naganathan, Sundar Ram; Fürthauer, Sebastian; Nishikawa, Masatoshi; Jülicher, Frank; Grill, Stephan W

    2014-01-01

    Many developmental processes break left–right (LR) symmetry with a consistent handedness. LR asymmetry emerges early in development, and in many species the primary determinant of this asymmetry has been linked to the cytoskeleton. However, the nature of the underlying chirally asymmetric cytoskeletal processes has remained elusive. In this study, we combine thin-film active chiral fluid theory with experimental analysis of the C. elegans embryo to show that the actomyosin cortex generates active chiral torques to facilitate chiral symmetry breaking. Active torques drive chiral counter-rotating cortical flow in the zygote, depend on myosin activity, and can be altered through mild changes in Rho signaling. Notably, they also execute the chiral skew event at the 4-cell stage to establish the C. elegans LR body axis. Taken together, our results uncover a novel, large-scale physical activity of the actomyosin cytoskeleton that provides a fundamental mechanism for chiral morphogenesis in development. DOI: http://dx.doi.org/10.7554/eLife.04165.001 PMID:25517077

  10. 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

  11. Assessment of immunoglobulin concentrates on thrombogenic activity by thrombin generation assay, prekallikrein activator assay, and size-exclusion chromatography.

    PubMed

    Seifner, Alexandra; Beck, Gerhard; Bayer, Patrick; Eichmeir, Stephanie; Lackner, Friedrich; Rögelsperger, Olga; Weber, Katharina; Wollein, Gabriele

    2014-02-01

    Immunoglobulin G (IgG) concentrates have recently been found to be contaminated with procoagulant impurities causing thromboembolic events (TEEs) in vivo. In this study the question was raised whether a thrombin generation assay (TGA) will be able to characterize IgG samples from the Austrian market with regard to their thrombogenic potential. A total of 44 IgG concentrates have been assayed by TGA employing pooled normal plasma and Factor (F)XI-deficient plasma (FXIdp). Furthermore, the prekallikrein activator assay including determination of blank values, size-exclusion chromatography, and further test systems required for batch release testing of IgG concentrates according to the European Pharmacopeia (Pharm. Eur.) were carried out. All samples complied with acceptance criteria stated in the Plarm. Eur. and/or prescribed by the marketing approval. One intravenous immunoglobulin (IVIG) involved in TEEs exceeded a threshold level of 350 nmol peak thrombin, which was not exceeded after change of manufacture and by all the other IVIGs tested. Two hyperimmune globulins revealed elevated peak thrombin levels of up to 810 nmol in FXI and up to 285 nmol in FXIdp. The study indicates that the TGA is able to reliably predict procoagulant activities probably associated with the presence of FXIa and potential thrombogenicity. Comparison of thrombin generation with product-specific acceptance criteria as well as variables from other test systems as amidolytic activity and molecular size can help to monitor IgG quality and manufacturing changes with regard to thrombogenicity. © 2013 American Association of Blood Banks.

  12. 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.

  13. 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

  14. Activity rhythm of Drosophila kept in complete darkness for 1300 generations.

    PubMed

    Imafuku, Michio; Haramura, Takashi

    2011-03-01

    Locomotor activity rhythms of dark stock flies of Drosophila melanogaster kept in complete darkness for 700 to 1340 generations were examined. The stock was established by the late Prof. S. Mori in November 1954 to investigate long-term effects of darkness on organisms. The activity of flies was recorded under three types of light conditions: DD after LD12:12, and DD after exposure to a 3.5 h (P3) or 7.5 h (P7) light pulse. In all of these conditions, the experimental dark flies exhibited clear circadian rhythms similar to those of control light flies. We compare our results with those of various studies on troglobites.

  15. 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.

  16. Activation of the NRF2 antioxidant program generates an imbalance in central carbon metabolism in cancer.

    PubMed

    Sayin, Volkan I; LeBoeuf, Sarah E; Singh, Simranjit X; Davidson, Shawn M; Biancur, Douglas; Guzelhan, Betul S; Alvarez, Samantha W; Wu, Warren L; Karakousi, Triantafyllia R; Zavitsanou, Anastasia Maria; Ubriaco, Julian; Muir, Alexander; Karagiannis, Dimitris; Morris, Patrick J; Thomas, Craig J; Possemato, Richard; Vander Heiden, Matthew G; Papagiannakopoulos, Thales

    2017-10-02

    During tumorigenesis, the high metabolic demand of cancer cells results in increased production of reactive oxygen species. To maintain oxidative homeostasis, tumor cells increase their antioxidant production through hyperactivation of the NRF2 pathway, which promotes tumor cell growth. Despite the extensive characterization of NRF2-driven metabolic rewiring, little is known about the metabolic liabilities generated by this reprogramming. Here, we show that activation of NRF2, in either mouse or human cancer cells, leads to increased dependency on exogenous glutamine through increased consumption of glutamate for glutathione synthesis and glutamate secretion by xc(-) antiporter system. Together, this limits glutamate availability for the tricarboxylic acid cycle and other biosynthetic reactions creating a metabolic bottleneck. Cancers with genetic or pharmacological activation of the NRF2 antioxidant pathway have a metabolic imbalance between supporting increased antioxidant capacity over central carbon metabolism, which can be therapeutically exploited.

  17. 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.

  18. Tandem surface microfluidic lithography and activation to generate patch pattern biospecific ligand and cell arrays.

    PubMed

    Pulsipher, Abigail; Yousaf, Muhammad N

    2010-03-16

    We report a rapid, inexpensive, and flexible methodology that combines microfluidic lithography and oxidative activation to pattern and chemically alter selective regions of SAMs on gold for subsequent chemoselective ligand immobilization. We demonstrate that PCC, a mild oxidant, can be used to convert hydroxyl-terminated SAMs to aldehydes and decorated with a variety of oxyamine-containing molecules. This strategy is compatible with cell culture and was employed to create a biospecific ligand platform for peptide-mediated, cell adhesion arrays. By using a number of different ligands and characterization tools, we showed that the generation of both cell patterning and ligand microarray patterning can be achieved. SAM formation, activation, ligand immobilization, and biospecific cell patterning are characterized by contact angle, cyclic voltammetry (CV), X-ray photoelectron spectroscopy (XPS) (Supporting Information), scanning electron microscopy (SEM), and fluorescence microscopy.

  19. Automatic generation of active coordinates for quantum dynamics calculations: Application to the dynamics of benzene photochemistry

    SciTech Connect

    Lasorne, Benjamin; Sicilia, Fabrizio; Bearpark, Michael J.; Robb, Michael A.; Worth, Graham A.; Blancafort, Lluis

    2008-03-28

    A new practical method to generate a subspace of active coordinates for quantum dynamics calculations is presented. These reduced coordinates are obtained as the normal modes of an analytical quadratic representation of the energy difference between excited and ground states within the complete active space self-consistent field method. At the Franck-Condon point, the largest negative eigenvalues of this Hessian correspond to the photoactive modes: those that reduce the energy difference and lead to the conical intersection; eigenvalues close to 0 correspond to bath modes, while modes with large positive eigenvalues are photoinactive vibrations, which increase the energy difference. The efficacy of quantum dynamics run in the subspace of the photoactive modes is illustrated with the photochemistry of benzene, where theoretical simulations are designed to assist optimal control experiments.

  20. 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.

  1. Generation of High Frequency Electric Field Activity by Turbulence in the Earth's Magnetotail

    NASA Astrophysics Data System (ADS)

    Stawarz, J. E.; Ergun, R.

    2013-12-01

    Bursty Bulk Flow (BBF) events, frequently observed in the magnetotail, carry significant energy and mass from the tail region at ~20 RE into the near-earth plasma sheet at ~10 RE, which is often referred to as the BBF 'braking region'. A number of possible channels are available for the transfer or dissipation of energy in BBF events including adiabatic heating of ions and electrons, the propagation of Alfvén waves out of the BBF braking region and into the auroral region, and energy dissipation within the braking region itself. This study investigates the generation of strong high frequency electric field activity observed within the braking region. A theory by which the large and small scales are coupled through a turbulent cascade of Alfvén waves, generated by the BBF braking event, is considered. At small kinetic spatial scales magnetic field aligned currents can be generated. These currents can be unstable to high frequency electrostatic waves, as well as, non-linear electrostatic structures such as double layers and electron phase space holes that are observed in the breaking region. The theoretical work is supported by observations from the THEMIS satellites. This work provides a possible mechanism for the dissipation of energy in turbulent plasma environments.

  2. 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

  3. 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.

  4. 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.

  5. 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.

  6. 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

  7. 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.

  8. 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.

  9. 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

  10. 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

  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. 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.

  14. Isolating early cortical generators of visual-evoked activity: a systems identification approach.

    PubMed

    Murphy, Jeremy W; Kelly, Simon P; Foxe, John J; Lalor, Edmund C

    2012-07-01

    The VESPA (visual-evoked spread spectrum analysis) method estimates the impulse response of the visual system using a continuously varying stimulus. It has been used recently to address both basic cognitive and neurophysiologic questions as well as those surrounding clinical populations. Although the components of the average VESPA response are highly reminiscent of the early components of the visual-evoked potential (VEP) when measured over midline occipital locations, the two responses are acquired in different ways and, thus, they cannot be regarded as being equivalent. To further characterize the relationship between the VESPA and the VEP and the generative mechanisms underlying them, we recorded EEG from 31 subjects in response to checkerboard-based VEP and VESPA stimuli. We found that, across subjects, the amplitudes of the VEP C1 component and the VESPA C1 component were highly correlated, whereas the VEP P1 and the VESPA P1 bore no statistical relationship. Furthermore, we found that C1 and P1 amplitudes were significantly correlated in the VESPA but not in the VEP. We believe these findings point to the presence of common generators underlying the VESPA C1 and the VEP C1. We argue further that the VESPA P1, in light of its strong relationship to the VESPA C1, likely reflects further activation of the same cortical generators. Given the lack of correlation between the VEP P1 and each of these three other components, it is likely that the underlying generators of this particular component are more varied and widespread, as suggested previously. We discuss the implications of these relationships for basic and clinical research using the VESPA and for the assessment of additive-evoked versus phase-reset contributions to the VEP.

  15. 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.

  16. Cathepsin L colocalizes with chromogranin a in chromaffin vesicles to generate active peptides.

    PubMed

    Biswas, Nilima; Rodriguez-Flores, Juan L; Courel, Maite; Gayen, Jiaur R; Vaingankar, Sucheta M; Mahata, Manjula; Torpey, Justin W; Taupenot, Laurent; O'Connor, Daniel T; Mahata, Sushil K

    2009-08-01

    Chromogranin A (CgA), the major soluble protein in chromaffin granules, is proteolytically processed to generate biologically active peptides including the catecholamine release inhibitory peptide catestatin. Here we sought to determine whether cysteine protease cathepsin L (CTSL), a novel enzyme for proteolytic processing of neuropeptides, acts like the well-established serine proteases [prohormone convertase (PC)1/3 or PC2] to generate catestatin by proteolytic processing of CgA. We found that endogenous CTSL colocalizes with CgA in the secretory vesicles of primary rat chromaffin cells. Transfection of PC12 cells with an expression plasmid encoding CTSL directed expression of CTSL toward secretory vesicles. Deconvolution fluorescence microscopy suggested greater colocalization of CTSL with CgA than the lysosomal marker LGP110. The overexpression of CTSL in PC12 cells caused cleavage of full-length CgA. CTSL also cleaved CgA in vitro, in time- and dose-dependent fashion, and specificity of the process was documented through E64 (thiol reagent) inhibition. Mass spectrometry on CTSL-digested recombinant CgA identified a catestatin-region peptide, corresponding to CgA(360-373). The pool of peptides generated from the CTSL cleavage of CgA inhibited nicotine-induced catecholamine secretion from PC12 cells. CTSL processing in the catestatin region was diminished by naturally occurring catestatin variants, especially Pro370Leu and Gly364Ser. Among the CTSL-generated peptides, a subset matched those found in the catestatin region in vivo. These findings indicate that CgA can be a substrate for the cysteine protease CTSL both in vitro and in cella, and their colocalization within chromaffin granules in cella suggests the likelihood of an enzyme/substrate relationship in vivo.

  17. TMEM16A channels generate Ca2+-activated Cl− currents in cerebral artery smooth muscle cells

    PubMed Central

    Thomas-Gatewood, Candice; Neeb, Zachary P.; Bulley, Simon; Adebiyi, Adebowale; Bannister, John P.; Leo, M. Dennis

    2011-01-01

    Transmembrane protein (TMEM)16A channels are recently discovered membrane proteins that display electrophysiological properties similar to classic Ca2+-activated Cl− (ClCa) channels in native cells. The molecular identity of proteins that generate ClCa currents in smooth muscle cells (SMCs) of resistance-size arteries is unclear. Similarly, whether cerebral artery SMCs generate ClCa currents is controversial. Here, using molecular biology and patch-clamp electrophysiology, we examined TMEM16A channel expression and characterized Cl− currents in arterial SMCs of resistance-size rat cerebral arteries. RT-PCR amplified transcripts for TMEM16A but not TMEM16B–TMEM16H, TMEM16J, or TMEM16K family members in isolated pure cerebral artery SMCs. Western blot analysis using an antibody that recognized recombinant (r)TMEM16A channels detected TMEM16A protein in cerebral artery lysates. Arterial surface biotinylation and immunofluorescence indicated that TMEM16A channels are located primarily within the arterial SMC plasma membrane. Whole cell ClCa currents in arterial SMCs displayed properties similar to those generated by rTMEM16A channels, including Ca2+ dependence, current-voltage relationship linearization by an elevation in intracellular Ca2+ concentration, a Nerstian shift in reversal potential induced by reducing the extracellular Cl− concentration, and a negative reversal potential shift when substituting extracellular I− for Cl−. A pore-targeting TMEM16A antibody similarly inhibited both arterial SMC ClCa and rTMEM16A currents. TMEM16A knockdown using small interfering RNA also inhibited arterial SMC ClCa currents. In summary, these data indicate that TMEM16A channels are expressed, insert into the plasma membrane, and generate ClCa currents in cerebral artery SMCs. PMID:21856902

  18. 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

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

    PubMed Central

    Schultz, Wolfram

    2015-01-01

    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 towards 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 for goal-directed behavior. The existence of neuronal planning activity in the amygdala suggests an important role for this structure in guiding behavior towards internally generated, distant goals. PMID:25622146

  20. 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.

  1. 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

  2. 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.

  3. New evidence for persistent impact-generated hydrothermal activity in the Miocene Ries impact structure, Germany

    NASA Astrophysics Data System (ADS)

    Arp, Gernot; Kolepka, Claudia; Simon, Klaus; Karius, Volker; Nolte, Nicole; Hansen, Bent T.

    2013-12-01

    The extent of impact-generated hydrothermal activity in the 24 km sized Ries impact structure has been controversially discussed. To date, mineralogical and isotopic investigations point to a restriction of hydrothermal activity to the impact-melt bearing breccias, specifically the crater-fill suevite. Here, we present new petrographic, geochemical, and isotopic data of postimpact carbonate deposits, which indicate a hydrothermal activity more extended than previously assumed. Specifically, carbonates of the Erbisberg, a spring mound located upon the inner crystalline ring of the crater, show travertine facies types not seen in any of the previously investigated sublacustrine soda lake spring mounds of the Ries basin. In particular, the streamer carbonates, which result from the encrustation of microbial filaments in subaerial spring effluents between 60 and 70 °C, are characteristic of a hydrothermal origin. While much of the primary geochemical and isotopic signatures in the mound carbonates have been obliterated by diagenesis, a postimpact calcite vein from brecciated gneiss of the subsurface crater floor revealed a flat rare earth element pattern with a clear positive Eu anomaly, indicating a hydrothermal fluid convection in the crater basement. Finally, the strontium isotope stratigraphic correlation of the travertine mound with the crater basin succession suggests a hydrothermal activity for about 250,000 yr after the impact, which would be much longer than previously assumed.

  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. Control of APP processing and Abeta generation level by BACE1 enzymatic activity and transcription.

    PubMed

    Li, Yu; Zhou, Weihui; Tong, Yigang; He, Guiqiong; Song, Weihong

    2006-02-01

    Deposition of amyloid beta protein (Abeta) is one of the characteristic features of Alzheimer's disease (AD) neuropathology. Beta-secretase, a beta-site APP cleaving enzyme 1 (BACE1), is essential for Abeta biosynthesis. Although inhibition of BACE1 is considered a valid therapeutic target for AD, the enzymatic dynamics of BACE1 in regulating APP processing and Abeta generation has not yet been fully defined. To examine this issue, tightly controlled inducible BACE1 gene expression was established in the neuronal cell line N2ABP1 and the non-neuronal cell line E2BP1 using an ecdysone-inducible system. The BACE1 protein level was increased in a time- and dosage-dependent manner in the inducible BACE1 stable cells by treatment with inducer ponasterone A. The generation of APP CTFbeta, the beta-secretase product, increased proportionally with the level of BACE1 protein expression. However, Abeta40/42 production sharply increased to the plateau level with a relatively small increase in BACE1 expression. Although further increasing BACE1 expression increased beta-secretase activity, it had no additional effect on Abeta production. Furthermore, we found that BACE1 mRNA levels and BACE1 promoter activity were significantly lower than APP mRNA levels and APP promoter activity. Our data demonstrate that lower BACE transcription is responsible for the minority of APP undergoing the amyloidogenic pathway and relatively lower Abeta production in the normal conditions, and that a slight increase in BACE1 can induce a dramatic elevation in Abeta production, indicating that the increase in BACE1 can potentially increase neuritic plaque formation in the pathological condition.

  7. 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

  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. Under What Conditions Are Embedded versus Generative Cognitive Strategy Activators Effective? Two Prescriptive Models for Designing Instruction.

    ERIC Educational Resources Information Center

    Darwazeh, Afnan N.

    In an embedded cognitive-strategy system, learners are forced to use a given cognitive strategy prepared by the instructor, designer, or researcher. In a generative cognitive-strategy system, learners are directed to generate a certain cognitive strategy activator by themselves. This is a review of experimental and theoretical studies conducted on…

  11. Solvent-free synthesis, coating and morphogenesis of conductive polymer materials through spontaneous generation of activated monomers.

    PubMed

    Muramatsu, Ryo; Oaki, Yuya; Kuwabara, Kento; Hayashi, Kosei; Imai, Hiroaki

    2014-10-14

    Synthesis, coating, and morphogenesis of conductive polymers were achieved on a variety of substrates through spontaneous generation of activated monomer vapors under ambient pressure and low temperature conditions. The present approach facilitates the generation of complex hierarchical morphologies and the conductive coating for improvement of electrochemical properties.

  12. 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.

  13. 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

  14. 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.

  15. 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

  16. 3-Hydroxykynurenine, a Tryptophan Metabolite Generated during the Infection, Is Active Against Trypanosoma cruzi.

    PubMed

    Knubel, Carolina P; Insfran, Constanza; Martinez, Fernando F; Diaz Lujan, Cintia; Fretes, Ricardo E; Theumer, Martin G; Cervi, Laura; Motran, Claudia C

    2017-07-13

    The antiparasitic activity of 3-hydroxykynurenine (3-HK), one of the major tryptophan catabolites of the kynurenine pathway, against both Trypanosoma cruzi evolutive forms that are important for human infection, trypomastigotes (Tps) and amastigotes (Am), possible targets in the parasite and the drug toxicity to mammalian cells have been investigated. 3-HK showed a potent activity against Am with IC50 values in the micromolar concentration range, while the IC50 values to cause Tps death was ∼6000-times higher, indicating that the replicative form present in the vertebrate hosts is much more susceptible to 3-HK than bloodstream Tps. In addition, 3-HK showed activity against Tps and Am, at concentrations that did not exhibit toxicity to mammalian cells. Ultrastructural analysis and flow cytometry studies indicated that Am and Tps mitochondrion and nuclei contain 3-HK targets. The potency and selectivity of 3-HK, which is generated during T. cruzi infection in human and mice, suggest that 3-HK may be a suitable candidate for drug research and development for Chagas disease.

  17. 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.

  18. 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.

  19. 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

  20. Combining generative and discriminative models for semantic segmentation of CT scans via active learning.

    PubMed

    Iglesias, Juan Eugenio; Konukoglu, Ender; Montillo, Albert; Tu, Zhuowen; Criminisi, Antonio

    2011-01-01

    This paper presents a new supervised learning framework for the efficient recognition and segmentation of anatomical structures in 3D computed tomography (CT), with as little training data as possible. Training supervised classifiers to recognize organs within CT scans requires a large number of manually delineated exemplar 3D images, which are very expensive to obtain. In this study, we borrow ideas from the field of active learning to optimally select a minimum subset of such images that yields accurate anatomy segmentation. The main contribution of this work is in designing a combined generative-discriminative model which: i) drives optimal selection of training data; and ii) increases segmentation accuracy. The optimal training set is constructed by finding unlabeled scans which maximize the disagreement between our two complementary probabilistic models, as measured by a modified version of the Jensen-Shannon divergence. Our algorithm is assessed on a database of 196 labeled clinical CT scans with high variability in resolution, anatomy, pathologies, etc. Quantitative evaluation shows that, compared with randomly selecting the scans to annotate, our method decreases the number of training images by up to 45%. Moreover, our generative model of body shape substantially increases segmentation accuracy when compared to either using the discriminative model alone or a generic smoothness prior (e.g. via a Markov Random Field).

  1. 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.

  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. Coherent combining of second-harmonic generators by active phase control of the fundamental waves.

    PubMed

    Odier, Alice; Durécu, Anne; Melkonian, Jean-Michel; Lombard, Laurent; Lefebvre, Michel; Bourdon, Pierre

    2017-08-15

    Coherent beam combining by active phase control could be useful for power scaling fiber-laser-pumped optical frequency converters. However, a fast phase modulator operating at the frequency-converted wavelength, a non-standard component, would be necessary. Fortunately, nonlinear conversion processes rely on a phase-matching condition allowing for indirect phase control using standard phase modulators. In this Letter, coherent combining of second-harmonic generators is demonstrated in both birefringent and quasi-phase-matching schemes in CW regime. Phase control operates at the fundamental wavelength, using all-fiber electro-optic modulators. An excellent beam combination is achieved with a residual phase error of λ/30 on the second-harmonic wave.

  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. 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.