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Sample records for activation profiles depending

  1. Mass spectrometric analysis of activity-dependent changes of neuropeptide profile in the snail, Helix pomatia.

    PubMed

    Pirger, Z; Lubics, A; Reglodi, D; Laszlo, Z; Mark, L; Kiss, T

    2010-12-01

    Terrestrial snails are able to transform themselves into inactivity ceasing their behavioral activity under unfavorable environmental conditions. In the present study, we report on the activity-dependent changes of the peptide and/or polypeptide profile in the brain and hemolymph of the snail, Helix pomatia, using MALDI TOF and quadrupole mass spectrometry. The present data indicate that the snails respond to low temperature by increasing or decreasing the output of selected peptides. Average mass spectra of the brain and hemolymph revealed numerous peaks predominantly present during the active state (19 and 10 peptides/polypeptides, respectively), while others were observed only during hibernation (11 and 13). However, there were peptides and/or polypeptides or their fragments present irrespective of the activity states (49 and 18). The intensity of fourteen peaks that correspond to previously identified neuropeptides varied in the brain of active snails compared to those of hibernating animals. Among those the intensity of eight peptides increased significantly in active animals while in hibernated animals the intensity of another six peptides increased significantly. A new peptide or peptide fragment at m/z 1110.7 was identified in a brain of the snail with the following suggested amino acid sequence: GSGASGSMPATTS. This peptide was found to be more abundant in active animals because the intensity of the peptide was significantly higher compared to hibernating animals. In summary, our results revealed substantial differences in the peptide/polypeptide profile of the brain and hemolymph of active and hibernating snails suggesting a possible contribution of peptides in the process of hibernation. Copyright © 2010 Elsevier Ltd. All rights reserved.

  2. INCLINATION-DEPENDENT ACTIVE GALACTIC NUCLEUS FLUX PROFILES FROM STRONG LENSING OF THE KERR SPACETIME

    SciTech Connect

    Chen, Bin; Dai, Xinyu; Baron, E.

    2013-01-10

    Recent quasar microlensing observations have constrained the X-ray emission sizes of quasars to be about 10 gravitational radii, one order of magnitude smaller than the optical emission sizes. Using a new ray-tracing code for the Kerr spacetime, we find that the observed X-ray flux is strongly influenced by the gravity field of the central black hole, even for observers at moderate inclination angles. We calculate inclination-dependent flux profiles of active galactic nuclei in the optical and X-ray bands by combining the Kerr lensing and projection effects for future reference. We further study the dependence of the X-ray-to-optical flux ratio on the inclination angle caused by differential lensing distortion of the X-ray and optical emission, assuming several corona geometries. The strong lensing X-ray-to-optical magnification ratio can change by a factor of {approx}10 for normal quasars in some cases, and a further factor of {approx}10 for broad absorption line (BAL) quasars and obscured quasars. Comparing our results with the observed distributions in normal and BAL quasars, we find that the inclination angle dependence of the magnification ratios can significantly change the X-ray-to-optical flux ratio distributions. In particular, the mean value of the spectrum slope parameter {alpha}{sub ox}, 0.3838log F {sub 2keV}/F {sub 2500A}, can differ by {approx}0.1-0.2 between normal and BAL quasars, depending on corona geometries, suggesting larger intrinsic absorptions in BAL quasars.

  3. Active dependency.

    PubMed

    Bornstein, R F

    1995-02-01

    Although dependency has long been associated with passivity, weakness, and submissiveness, a review of the empirical literature reveals that, in certain situations and settings, dependent persons actually exhibit a variety of active, assertive behaviors. In this article, I: a) trace the historical roots of the dependency-passivity link; b) review empirical studies from developmental, social, and clinical psychology which indicate that, in certain circumstances, dependency is associated with active, assertive behavior on the part of the dependent person; c) offer an alternative conceptual model of dependency that accounts for the entire range of behaviors-both passive and active-that are exhibited by the dependent person; and d) discuss the diagnostic and therapeutic implications of this alternative conceptual model of dependency.

  4. Analysis of PBase Binding Profile Indicates an Insertion Target Selection Mechanism Dependent on TTAA, But Not Transcriptional Activity

    PubMed Central

    Yang, Dong; Liao, Ruiqi; Zheng, Yun; Sun, Ling; Xu, Tian

    2016-01-01

    Transposons and retroviruses are important pathogenic agents and tools for mutagenesis and transgenesis. Insertion target selection is a key feature for a given transposon or retrovirus. The piggyBac (PB) transposon is highly active in mice and human cells, which has a much better genome-wide distribution compared to the retrovirus and P-element. However, the underlying reason is not clear. Utilizing a tagged functional PB transposase (PBase), we were able to conduct genome-wide profiling for PBase binding sites in the mouse genome. We have shown that PBase binding mainly depends on the distribution of the tetranucleotide TTAA, which is not affected by the presence of PB DNA. Furthermore, PBase binding is negatively influenced by the methylation of CG sites in the genome. Analysis of a large collection of PB insertions in mice has revealed an insertion profile similar to the PBase binding profile. Interestingly, this profile is not correlated with transcriptional active genes in the genome or transcriptionally active regions within a transcriptional unit. This differs from what has been previously shown for P-element and retroviruses insertions. Our study provides an explanation for PB's genome-wide insertion distribution and also suggests that PB target selection relies on a new mechanism independent of active transcription and open chromatin structure. PMID:27570481

  5. Palmitoylome profiling reveals S-palmitoylation–dependent antiviral activity of IFITM3

    PubMed Central

    Yount, Jacob S; Moltedo, Bruno; Yang, Yu-Ying; Charron, Guillaume; Moran, Thomas M; López, Carolina B; Hang, Howard C

    2010-01-01

    Identification of immune effectors and the post-translational modifications that control their activity is essential for dissecting mechanisms of immunity. Here we demonstrate that the antiviral activity of interferon-induced transmembrane protein 3 (IFITM3) is post-translationally regulated by S-palmitoylation. Large-scale profiling of palmitoylated proteins in a dendritic cell line using a chemical reporter strategy revealed over 150 lipid-modified proteins with diverse cellular functions, including innate immunity. We discovered that S-palmitoylation of IFITM3 on membrane-proximal cysteines controls its clustering in membrane compartments and its antiviral activity against influenza virus. The sites of S-palmitoylation are highly conserved among the IFITM family of proteins in vertebrates, which suggests that S-palmitoylation of these immune effectors may be an ancient post-translational modification that is crucial for host resistance to viral infections. The S-palmitoylation and clustering of IFITM3 will be important for elucidating its mechanism of action and for the design of antiviral therapeutics. PMID:20601941

  6. Power output in vertical jumps: does optimum loading depend on activity profiles?

    PubMed

    Pazin, Nemanja; Berjan, Bobana; Nedeljkovic, Aleksandar; Markovic, Goran; Jaric, Slobodan

    2013-03-01

    The previously proposed maximum dynamic output hypothesis (MDO: i.e. the optimum load for maximizing the power output during jumping is one's own body) was tested on individuals of various activity profiles. Forty males (10 strength-trained athletes, 10 speed-trained athletes, 10 physically active non-athletes, and 10 sedentary individuals) performed different vertical jumps on a force plate while a pulley system was used to either reduce or increase the subject's body weight by 10-30 %. As expected, an increase in external loading resulted in a significant increase (p < 0.001) in force output and a concomitant decrease of peak jumping velocity in all groups of participants. The main finding, however, was that all groups revealed the maximum peak and mean power output at approximately the subjects' own body weight although their weight represented prominently different percentage of their maximum dynamic strength. While a significant (p < 0.05), albeit moderate, 'group × load' interaction in one jump was observed for the peak power output, the individual optimum load for maximizing the power output number did not differ among the groups. Although apparently further research on various types of movements is needed, the present results provide, so far, the strongest support of the MDO hypothesis.

  7. Ca2+/Calmodulin-Dependent Kinase Kinase α Is Expressed by Monocytic Cells and Regulates the Activation Profile

    PubMed Central

    Guest, Christopher B.; Deszo, Eric L.; Hartman, Matthew E.; York, Jason M.; Kelley, Keith W.; Freund, Gregory G.

    2008-01-01

    Macrophages are capable of assuming numerous phenotypes in order to adapt to endogenous and exogenous challenges but many of the factors that regulate this process are still unknown. We report that Ca2+/calmodulin-dependent kinase kinase α (CaMKKα) is expressed in human monocytic cells and demonstrate that its inhibition blocks type-II monocytic cell activation and promotes classical activation. Affinity chromatography with paramagnetic beads isolated an approximately 50 kDa protein from nuclear lysates of U937 human monocytic cells activated with phorbol-12-myristate-13-acetate (PMA). This protein was identified as CaMKKα by mass spectrometry and Western analysis. The function of CaMKKα in monocyte activation was examined using the CaMKKα inhibitors (STO-609 and forskolin) and siRNA knockdown. Inhibition of CaMKKα, enhanced PMA-dependent CD86 expression and reduced CD11b expression. In addition, inhibition was associated with decreased translocation of CaMKKα to the nucleus. Finally, to further examine monocyte activation profiles, TNFα and IL-10 secretion were studied. CaMKKα inhibition attenuated PMA-dependent IL-10 production and enhanced TNFα production indicating a shift from type-II to classical monocyte activation. Taken together, these findings indicate an important new role for CaMKKα in the differentiation of monocytic cells. PMID:18270593

  8. Probing kinase and phosphatase activities of two-component systems in vivo with concentration-dependent phosphorylation profiling

    PubMed Central

    Gao, Rong; Stock, Ann M.

    2013-01-01

    Quantitative analyses of protein concentrations, modifications and activities in their native environments are playing an increasingly vital role in unraveling the general principles underlying signal transduction pathways. The prevalent bacterial two-component systems (TCSs) use a central phosphotransfer for signaling; however, in vivo characterization of the kinase and phosphatase activities of TCS proteins is often limited by traditional transcriptional reporter assays and complicated by simultaneous actions of multiple TCS activities. Here, we report a strategy that combines concentration-dependent phosphorylation profiling and mathematical modeling to characterize the cellular activities of the archetype Escherichia coli PhoR/PhoB system. Phosphorylation of the response regulator (RR) PhoB has been found to be dependent on the total concentrations of PhoB/PhoR and saturated at high concentrations. The relationship between RR phosphorylation and total concentrations has been defined by the modeling of the kinase and phosphatase reactions and quantified to derive the biochemical parameters of the PhoR/PhoB system in vivo. In a further test of this approach on a PhoB mutant, PhoBF20D, it proved highly effective in exploring the mechanistic differences of TCSs that are not revealed by traditional reporter assays. Measurement of biochemical parameters for PhoBF20D led to the discovery that a weaker interaction between the histidine sensor kinase and RR could result in a higher and nonrobust phosphorylation due to diminished phosphatase activities. PMID:23267085

  9. Profiling of Sox4-dependent transcriptome in skin links tumour suppression and adult stem cell activation.

    PubMed

    Foronda, Miguel; Morgado-Palacin, Lucia; Gómez-López, Gonzalo; Domínguez, Orlando; Pisano, David G; Blasco, Maria A

    2015-12-01

    Adult stem cells (ASCs) reside in specific niches in a quiescent state in adult mammals. Upon specific cues they become activated and respond by self-renewing and differentiating into newly generated specialised cells that ensure appropriate tissue fitness. ASC quiescence also serves as a tumour suppression mechanism by hampering cellular transformation and expansion (White AC et al., 2014). Some genes restricted to early embryonic development and adult stem cell niches are often potent modulators of stem cell quiescence, and derailed expression of these is commonly associated to cancer (Vervoort SJ et al., 2013). Among them, it has been shown that recommissioned Sox4 expression facilitates proliferation, survival and migration of malignant cells. By generating a conditional Knockout mouse model in stratified epithelia (Sox4 (cKO) mice), we demonstrated a delayed plucking-induced Anagen in the absence of Sox4. Skin global transcriptome analysis revealed a prominent defect in the induction of transcriptional networks that control hair follicle stem cell (HFSC) activation such as those regulated by Wnt/Ctnnb1, Shh, Myc or Sox9, cell cycle and DNA damage response-associated pathways. Besides, Sox4 (cKO) mice are resistant to skin carcinogenesis, thus linking Sox4 to both normal and pathological HFSC activation (Foronda M et al., 2014). Here we provide additional details on the analysis of Sox4-regulated transcriptome in Telogen and Anagen skin. The raw and processed microarray data is deposited in GEO under GSE58155.

  10. Macrophages from the synovium of active rheumatoid arthritis exhibit an activin A-dependent pro-inflammatory profile.

    PubMed

    Soler Palacios, Blanca; Estrada-Capetillo, Lizbeth; Izquierdo, Elena; Criado, Gabriel; Nieto, Concha; Municio, Cristina; González-Alvaro, Isidoro; Sánchez-Mateos, Paloma; Pablos, Jose Luis; Corbí, Angel L; Puig-Kröger, Amaya

    2015-02-01

    Rheumatoid arthritis (RA) is a chronic inflammatory disease whose pathogenesis and severity correlates with the presence of macrophage-derived pro-inflammatory cytokines within the inflamed synovium. Macrophage-derived cytokines fuel the pathological processes in RA and are targets of clinically successful therapies. However, although macrophage polarization determines cytokine production, the polarization state of macrophages in RA joints remains poorly defined. To dissect the molecular basis for the tissue-damaging effects of macrophages in RA joints, we undertook the phenotypic and transcriptomic characterization of ex vivo isolated CD14(+) RA synovial fluid (RA-SF) macrophages. Flow cytometry and gene profiling indicated that RA-SF macrophages express pro-inflammatory polarization markers (MMP12, EGLN3, CCR2), lack expression of markers associated with homeostatic and anti-inflammatory polarization (IGF1, HTR2B) and exhibit a transcriptomic profile that resembles the activin A-dependent gene signature of pro-inflammatory in vitro-generated macrophages. In fact, high levels of Smad-activating activin A were found in RA-SF and, accordingly, the Smad signalling pathway was activated in ex vivo-isolated RA-SF macrophages. In vitro experiments on monocytes and macrophages indicated that RA-SF promoted the acquisition of pro-inflammatory markers (INHBA, MMP12, EGLN3, CCR2) but led to a significant reduction in the expression of genes associated with homeostasis and inflammation resolution (FOLR2, SERPINB2, IGF1, CD36), thus confirming the pro-inflammatory polarization ability of RA-SF. Importantly, the macrophage-polarizing ability of RA-SF was inhibited by an anti-activin A-neutralizing antibody, thus demonstrating that activin A mediates the pro-inflammatory macrophage-polarizing ability of RA-SF. Moreover, and in line with these findings, multicolour immunofluorescence evidenced that macrophages within RA synovial membranes (RA-SM) also express pro

  11. Profiling the MAPK/ERK dependent and independent activity regulated transcriptional programs in the murine hippocampus in vivo

    PubMed Central

    Blüthgen, Nils; van Bentum, Mirjam; Merz, Barbara; Kuhl, Dietmar; Hermey, Guido

    2017-01-01

    Activity-dependent alteration of the transcriptional program is central for shaping neuronal connectivity. Constitutively expressed transcription factors orchestrate the initial response to neuronal stimulation and serve as substrates for second messenger-regulated kinase signalling cascades. The mitogen-activated protein kinase ERK conveys signalling from the synapse to the nucleus but its genetic signature following neuronal activity has not been revealed. The goal of the present study was to identify ERK dependent and independent activity regulated transcriptional programs in the murine hippocampus. We used generalized seizures combined with the pharmacological intervention of MEK activation as an in vivo model to determine the complete transcriptional program initiated by ERK after neuronal activity. Our survey demonstrates that the induction of a large number of activity-regulated genes, including Arc/Arg3.1, Arl5b, Gadd45b, Homer1, Inhba and Zwint, is indeed dependent on ERK phosphorylation. In contrast, expression of a small group of genes, including Npas4, Arl4d, Errfi1, and Rgs2, is only partially dependent or completely independent (Ppp1r15a) of this signalling pathway. Among the identified transcripts are long non-coding (lnc) RNAs and induction of LincPint and splice variants of NEAT1 are ERK dependent. Our survey provides a comprehensive analysis of the transcriptomic response conveyed by ERK signalling in the hippocampus. PMID:28349920

  12. Phenotypical and functional profiles of natural killer cells exhibiting matrix metalloproteinase-mediated CD16 cleavage after anti-HIV antibody-dependent activation.

    PubMed

    Tang, C-C; Isitman, G; Bruneau, J; Tremblay, C; Bernard, N F; Kent, S J; Parsons, M S

    2015-08-01

    Natural killer (NK) cell-mediated antibody-dependent cellular cytotoxicity (ADCC) has been linked to protection from HIV infection and slower progression towards AIDS. However, antibody-dependent activation of NK cells results in phenotypical alterations similar to those observed on NK cells from individuals with progressive HIV infection. Activation of NK cells induces matrix metalloproteinase (MMP)-mediated cleavage of cell surface CD16. In the present study we assessed the phenotype and functional profile of NK cells exhibiting post-activation MMP-mediated CD16 cleavage. We found that NK cells achieving the highest levels of activation during stimulation exhibit the most profound decreases in CD16 expression. Further, we observed that educated KIR3DL1(+) NK cells from human leucocyte antigen (HLA)-Bw4-carrying donors exhibit larger decreases in CD16 expression post-activation than the KIR3DL1(-) NK cell subset containing cells educated via other inhibitory receptor/ligand combinations and non-educated NK cells. Lastly, we assessed the ex-vivo expression of CD16 on educated KIR3DL1(+) NK cells and the KIR3DL1(-) NK cell subset from HLA-Bw4-carrying HIV-uninfected and HIV-infected donors. Suggestive of in-vivo activation of KIR3DL1(+) NK cells during HIV infection, CD16 expression was higher on KIR3DL1(+) than KIR3DL1(-) NK cells in uninfected donors but similar on both subsets in HIV-infected donors. These results are discussed in the context of how they may assist with understanding HIV disease progression and the design of immunotherapies that utilize antibody-dependent NK cell responses.

  13. Profiling dose-dependent activation of p53-mediated signaling pathways by chemicals with distinct mechanisms of DNA damage.

    PubMed

    Clewell, Rebecca A; Sun, Bin; Adeleye, Yeyejide; Carmichael, Paul; Efremenko, Alina; McMullen, Patrick D; Pendse, Salil; Trask, O J; White, Andy; Andersen, Melvin E

    2014-11-01

    As part of a larger effort to provide proof-of-concept in vitro-only risk assessments, we have developed a suite of high-throughput assays for key readouts in the p53 DNA damage response toxicity pathway: double-strand break DNA damage (p-H2AX), permanent chromosomal damage (micronuclei), p53 activation, p53 transcriptional activity, and cell fate (cell cycle arrest, apoptosis, micronuclei). Dose-response studies were performed with these protein and cell fate assays, together with whole genome transcriptomics, for three prototype chemicals: etoposide, quercetin, and methyl methanesulfonate. Data were collected in a human cell line expressing wild-type p53 (HT1080) and results were confirmed in a second p53 competent cell line (HCT 116). At chemical concentrations causing similar increases in p53 protein expression, p53-mediated protein expression and cellular processes showed substantial chemical-specific differences. These chemical-specific differences in the p53 transcriptional response appear to be determined by augmentation of the p53 response by co-regulators. More importantly, dose-response data for each of the chemicals indicate that the p53 transcriptional response does not prevent micronuclei induction at low concentrations. In fact, the no observed effect levels and benchmark doses for micronuclei induction were less than or equal to those for p53-mediated gene transcription regardless of the test chemical, indicating that p53's post-translational responses may be more important than transcriptional activation in the response to low dose DNA damage. This effort demonstrates the process of defining key assays required for a pathway-based, in vitro-only risk assessment, using the p53-mediated DNA damage response pathway as a prototype.

  14. Myofilament length dependent activation.

    PubMed

    de Tombe, Pieter P; Mateja, Ryan D; Tachampa, Kittipong; Ait Mou, Younss; Farman, Gerrie P; Irving, Thomas C

    2010-05-01

    The Frank-Starling law of the heart describes the interrelationship between end-diastolic volume and cardiac ejection volume, a regulatory system that operates on a beat-to-beat basis. The main cellular mechanism that underlies this phenomenon is an increase in the responsiveness of cardiac myofilaments to activating Ca(2+) ions at a longer sarcomere length, commonly referred to as myofilament length-dependent activation. This review focuses on what molecular mechanisms may underlie myofilament length dependency. Specifically, the roles of inter-filament spacing, thick and thin filament based regulation, as well as sarcomeric regulatory proteins are discussed. Although the "Frank-Starling law of the heart" constitutes a fundamental cardiac property that has been appreciated for well over a century, it is still not known in muscle how the contractile apparatus transduces the information concerning sarcomere length to modulate ventricular pressure development.

  15. Myofilament length dependent activation

    SciTech Connect

    de Tombe, Pieter P.; Mateja, Ryan D.; Tachampa, Kittipong; Mou, Younss Ait; Farman, Gerrie P.; Irving, Thomas C.

    2010-05-25

    The Frank-Starling law of the heart describes the interrelationship between end-diastolic volume and cardiac ejection volume, a regulatory system that operates on a beat-to-beat basis. The main cellular mechanism that underlies this phenomenon is an increase in the responsiveness of cardiac myofilaments to activating Ca{sup 2+} ions at a longer sarcomere length, commonly referred to as myofilament length-dependent activation. This review focuses on what molecular mechanisms may underlie myofilament length dependency. Specifically, the roles of inter-filament spacing, thick and thin filament based regulation, as well as sarcomeric regulatory proteins are discussed. Although the 'Frank-Starling law of the heart' constitutes a fundamental cardiac property that has been appreciated for well over a century, it is still not known in muscle how the contractile apparatus transduces the information concerning sarcomere length to modulate ventricular pressure development.

  16. Frequency dependence of pulsar integrated profiles

    SciTech Connect

    Thorsett, S.E. )

    1991-08-01

    The dependence of component separation on observing frequency has been studied for seven pulsars that exhibit double- or multiple-component average profiles. In each case, a review of all available data shows a smooth variation of given form. No evidence is found for a 'break frequency' at which the separation behavior discretely changes. It is argued that previous reports of such a discontinuity are due to insufficiently sampled data together with a prejudice toward pure power-law functional behaviors. The absence of such a break has implications for theories of the pulsar emission mechanism and of the propagation of radio waves in the pulsar magnetosphere. 44 refs.

  17. miRNA profiling during antigen-dependent T cell activation: A role for miR-132-3p.

    PubMed

    Gutiérrez-Vázquez, Cristina; Rodríguez-Galán, Ana; Fernández-Alfara, Marcos; Mittelbrunn, María; Sánchez-Cabo, Fátima; Martínez-Herrera, Dannys Jorge; Ramírez-Huesca, Marta; Pascual-Montano, Alberto; Sánchez-Madrid, Francisco

    2017-06-14

    microRNAs (miRNAs) are tightly regulated during T lymphocyte activation to enable the establishment of precise immune responses. Here, we analyzed the changes of the miRNA profiles of T cells in response to activation by cognate interaction with dendritic cells. We also studied mRNA targets common to miRNAs regulated in T cell activation. pik3r1 gene, which encodes the regulatory subunits of PI3K p50, p55 and p85, was identified as target of miRNAs upregulated after T cell activation. Using 3'UTR luciferase reporter-based and biochemical assays, we showed the inhibitory relationship between miR-132-3p upregulation and expression of the pik3r1 gene. Our results indicate that specific miRNAs whose expression is modulated during T cell activation might regulate PI3K signaling in T cells.

  18. Human serum metabolic profiles are age dependent

    PubMed Central

    Yu, Zhonghao; Zhai, Guangju; Singmann, Paula; He, Ying; Xu, Tao; Prehn, Cornelia; Römisch-Margl, Werner; Lattka, Eva; Gieger, Christian; Soranzo, Nicole; Heinrich, Joachim; Standl, Marie; Thiering, Elisabeth; Mittelstraß, Kirstin; Wichmann, Heinz-Erich; Peters, Annette; Suhre, Karsten; Li, Yixue; Adamski, Jerzy; Spector, Tim D; Illig, Thomas; Wang-Sattler, Rui

    2012-01-01

    Understanding the complexity of aging is of utmost importance. This can now be addressed by the novel and powerful approach of metabolomics. However, to date, only a few metabolic studies based on large samples are available. Here, we provide novel and specific information on age-related metabolite concentration changes in human homeostasis. We report results from two population-based studies: the KORA F4 study from Germany as a discovery cohort, with 1038 female and 1124 male participants (32–81 years), and the TwinsUK study as replication, with 724 female participants. Targeted metabolomics of fasting serum samples quantified 131 metabolites by FIA-MS/MS. Among these, 71/34 metabolites were significantly associated with age in women/men (BMI adjusted). We further identified a set of 13 independent metabolites in women (with P values ranging from 4.6 × 10−04 to 7.8 × 10−42, αcorr = 0.004). Eleven of these 13 metabolites were replicated in the TwinsUK study, including seven metabolite concentrations that increased with age (C0, C10:1, C12:1, C18:1, SM C16:1, SM C18:1, and PC aa C28:1), while histidine decreased. These results indicate that metabolic profiles are age dependent and might reflect different aging processes, such as incomplete mitochondrial fatty acid oxidation. The use of metabolomics will increase our understanding of aging networks and may lead to discoveries that help enhance healthy aging. PMID:22834969

  19. Frequency-dependent spatiotemporal profiles of visual responses recorded with subdural ECoG electrodes in awake monkeys: Differences between high- and low-frequency activity.

    PubMed

    Takaura, Kana; Tsuchiya, Naotsugu; Fujii, Naotaka

    2016-01-01

    Electrocorticography (ECoG) constitutes a powerful and promising neural recording modality in humans and animals. ECoG signals are often decomposed into several frequency bands, among which the so-called high-gamma band (80-250Hz) has been proposed to reflect local cortical functions near the cortical surface below the ECoG electrodes. It is typically assumed that the lower the frequency bands, the lower the spatial resolution of the signals; thus, there is not much to gain by analyzing the event-related changes of the ECoG signals in the lower-frequency bands. However, differences across frequency bands have not been systematically investigated. To address this issue, we recorded ECoG activity from two awake monkeys performing a retinotopic mapping task. We characterized the spatiotemporal profiles of the visual responses in the time-frequency domain. We defined the preferred spatial position, receptive field (RF), and response latencies of band-limited power (BLP) (i.e., alpha [3.9-11.7Hz], beta [15.6-23.4Hz], low [30-80Hz] and high [80-250Hz] gamma) for each electrode and compared them across bands and time-domain visual evoked potentials (VEPs). At the population level, we found that the spatial preferences were comparable across bands and VEPs. The high-gamma power showed a smaller RF than the other bands and VEPs. The response latencies for the alpha band were always longer than the latencies for the other bands and fastest in VEPs. Comparing the response profiles in both space and time for each cortical region (V1, V4+, and TEO/TE) revealed regional idiosyncrasies. Although the latencies of visual responses in the beta, low-, and high-gamma bands were almost identical in V1 and V4+, beta and low-gamma BLP occurred about 17ms earlier than high-gamma power in TEO/TE. Furthermore, TEO/TE exhibited a unique pattern in the spatial response profile: the alpha and high-gamma responses tended to prefer the foveal regions, whereas the beta and low-gamma responses

  20. Radiosensitivity profiles from a panel of ovarian cancer cell lines exhibiting genetic alterations in p53 and disparate DNA-dependent protein kinase activities

    SciTech Connect

    Langland, Gregory T.; Yannone, Steven M.; Langland, Rachel A.; Nakao, Aki; Guan, Yinghui; Long, Sydney B.T.; Vonguyen, Lien; Chen, David J.; Gray, Joe W; Chen, Fanqing

    2009-09-07

    The variability of radiation responses in ovarian tumors and tumor-derived cell lines is poorly understood. Since both DNA repair capacity and p53 status can significantly alter radiation sensitivity, we evaluated these factors along with radiation sensitivity in a panel of sporadic human ovarian carcinoma cell lines. We observed a gradation of radiation sensitivity among these sixteen lines, with a five-fold difference in the LD50 between the most radiosensitive and the most radioresistant cells. The DNA-dependent protein kinase (DNA-PK) is essential for the repair of radiation induced DNA double-strand breaks in human somatic cells. Therefore, we measured gene copy number, expression levels, protein abundance, genomic copy and kinase activity for DNA-PK in all of our cell lines. While there were detectable differences in DNA-PK between the cell lines, there was no clear correlation with any of these differences and radiation sensitivity. In contrast, p53 function as determined by two independent methods, correlated well with radiation sensitivity, indicating p53 mutant ovarian cancer cells are typically radioresistant relative to p53 wild-type lines. These data suggest that the activity of regulatory molecules such as p53 may be better indicators of radiation sensitivity than DNA repair enzymes such as DNAPK in ovarian cancer.

  1. Radiosensitivity profiles from a panel of ovarian cancer cell lines exhibiting genetic alterations in p53 and disparate DNA-dependent protein kinase activities

    PubMed Central

    Langland, Gregory T.; Yannone, Steven M.; Langland, Rachel A.; Nakao, Aki; Guan, Yinghui; Long, Sydney B.T.; Vonguyen, Lien; Chen, David J.; Gray, Joe W.; Chen, Fanqing

    2010-01-01

    The variability of radiation responses in ovarian tumors and tumor-derived cell lines is poorly understood. Since both DNA repair capacity and p53 status can significantly alter radiation sensitivity, we evaluated these factors along with radiation sensitivity in a panel of sporadic human ovarian carcinoma cell lines. We observed a gradation of radiation sensitivity among these sixteen lines, with a five-fold difference in the LD50 between the most radiosensitive and the most radioresistant cells. The DNA-dependent protein kinase (DNA-PK) is essential for the repair of radiation induced DNA double-strand breaks in human somatic cells. Therefore, we measured gene copy number, expression levels, protein abundance, genomic copy and kinase activity for DNA-PK in all of our cell lines. While there were detectable differences in DNA-PK between the cell lines, there was no clear correlation with any of these differences and radiation sensitivity. In contrast, p53 function as determined by two independent methods, correlated well with radiation sensitivity, indicating p53 mutant ovarian cancer cells are typically radioresistant relative to p53 wild-type lines. These data suggest that the activity of regulatory molecules such as p53 may be better indicators of radiation sensitivity than DNA repair enzymes such as DNA-PK in ovarian cancer. PMID:20204287

  2. Concentration profiles near an activated enzyme.

    PubMed

    Park, Soohyung; Agmon, Noam

    2008-09-25

    When a resting enzyme is activated, substrate concentration profile evolves in its vicinity, ultimately tending to steady state. We use modern theories for many-body effects on diffusion-influenced reactions to derive approximate analytical expressions for the steady-state profile and the Laplace transform of the transient concentration profiles. These show excellent agreement with accurate many-particle Brownian-dynamics simulations for the Michaelis-Menten kinetics. The steady-state profile has a hyperbolic dependence on the distance of the substrate from the enzyme, albeit with a prefactor containing the complexity of the many-body effects. These are most conspicuous for the substrate concentration at the surface of the enzyme. It shows an interesting transition as a function of the enzyme turnover rate. When it is high, the contact concentration decays monotonically to steady state. However, for slow turnover it is nonmonotonic, showing a minimum due to reversible substrate binding, then a maximum due to diffusion of new substrate toward the enzyme, and finally decay to steady state. Under certain conditions one can obtain a good estimate for the critical value of the turnover rate constant at the transition.

  3. Transcript profile analyses of maize silks reveal effective activation of genes involved in microtubule-based movement, ubiquitin-dependent protein degradation, and transport in the pollination process.

    PubMed

    Xu, Xiao Hui; Wang, Fang; Chen, Hao; Sun, Wei; Zhang, Xian Sheng

    2013-01-01

    Pollination is the first crucial step of sexual reproduction in flowering plants, and it requires communication and coordination between the pollen and the stigma. Maize (Zea mays) is a model monocot with extraordinarily long silks, and a fully sequenced genome, but little is known about the mechanism of its pollen-stigma interactions. In this study, the dynamic gene expression of silks at four different stages before and after pollination was analyzed. The expression profiles of immature silks (IMS), mature silks (MS), and silks at 20 minutes and 3 hours after pollination (20MAP and 3HAP, respectively) were compared. In total, we identified 6,337 differentially expressed genes in silks (SDEG) at the four stages. Among them, the expression of 172 genes were induced upon pollination, most of which participated in RNA binding, processing and transcription, signal transduction, and lipid metabolism processes. Genes in the SDEG dataset could be divided into 12 time-course clusters according to their expression patterns. Gene Ontology (GO) enrichment analysis revealed that many genes involved in microtubule-based movement, ubiquitin-mediated protein degradation, and transport were predominantly expressed at specific stages, indicating that they might play important roles in the pollination process of maize. These results add to current knowledge about the pollination process of grasses and provide a foundation for future studies on key genes involved in the pollen-silk interaction in maize.

  4. Transcript Profile Analyses of Maize Silks Reveal Effective Activation of Genes Involved in Microtubule-Based Movement, Ubiquitin-Dependent Protein Degradation, and Transport in the Pollination Process

    PubMed Central

    Chen, Hao; Sun, Wei; Zhang, Xian Sheng

    2013-01-01

    Pollination is the first crucial step of sexual reproduction in flowering plants, and it requires communication and coordination between the pollen and the stigma. Maize (Zea mays) is a model monocot with extraordinarily long silks, and a fully sequenced genome, but little is known about the mechanism of its pollen–stigma interactions. In this study, the dynamic gene expression of silks at four different stages before and after pollination was analyzed. The expression profiles of immature silks (IMS), mature silks (MS), and silks at 20 minutes and 3 hours after pollination (20MAP and 3HAP, respectively) were compared. In total, we identified 6,337 differentially expressed genes in silks (SDEG) at the four stages. Among them, the expression of 172 genes were induced upon pollination, most of which participated in RNA binding, processing and transcription, signal transduction, and lipid metabolism processes. Genes in the SDEG dataset could be divided into 12 time-course clusters according to their expression patterns. Gene Ontology (GO) enrichment analysis revealed that many genes involved in microtubule-based movement, ubiquitin-mediated protein degradation, and transport were predominantly expressed at specific stages, indicating that they might play important roles in the pollination process of maize. These results add to current knowledge about the pollination process of grasses and provide a foundation for future studies on key genes involved in the pollen–silk interaction in maize. PMID:23301084

  5. Expression and activity profiles of DPP IV/CD26 and NEP/CD10 glycoproteins in the human renal cancer are tumor-type dependent

    PubMed Central

    2010-01-01

    Background Cell-surface glycoproteins play critical roles in cell-to-cell recognition, signal transduction and regulation, thus being crucial in cell proliferation and cancer etiogenesis and development. DPP IV and NEP are ubiquitous glycopeptidases closely linked to tumor pathogenesis and development, and they are used as markers in some cancers. In the present study, the activity and protein and mRNA expression of these glycoproteins were analysed in a subset of clear-cell (CCRCC) and chromophobe (ChRCC) renal cell carcinomas, and in renal oncocytomas (RO). Methods Peptidase activities were measured by conventional enzymatic assays with fluorogen-derived substrates. Gene expression was quantitatively determined by qRT-PCR and membrane-bound protein expression and distribution analysis was performed by specific immunostaining. Results The activity of both glycoproteins was sharply decreased in the three histological types of renal tumors. Protein and mRNA expression was strongly downregulated in tumors from distal nephron (ChRCC and RO). Moreover, soluble DPP IV activity positively correlated with the aggressiveness of CCRCCs (higher activities in high grade tumors). Conclusions These results support the pivotal role for DPP IV and NEP in the malignant transformation pathways and point to these peptidases as potential diagnostic markers. PMID:20459800

  6. Crack depth profiling using guided wave angle dependent reflectivity

    SciTech Connect

    Volker, Arno Pahlavan, Lotfollah Blacquiere, Gerrit

    2015-03-31

    Tomographic corrosion monitoring techniques have been developed, using two rings of sensors around the circumference of a pipe. This technique is capable of providing a detailed wall thickness map, however this might not be the only type of structural damage. Therefore this concept is expanded to detect and size cracks and small corrosion defects like root corrosion. The expanded concept uses two arrays of guided-wave transducers, collecting both reflection and transmission data. The data is processed such that the angle-dependent reflectivity is obtained without using a baseline signal of a defect-free situation. The angle-dependent reflectivity is the input of an inversion scheme that calculates a crack depth profile. From this profile, the depth and length of the crack can be determined. Preliminary experiments show encouraging results. The depth sizing accuracy is in the order of 0.5 mm.

  7. Computational aspects of speed-dependent Voigt profiles

    NASA Astrophysics Data System (ADS)

    Schreier, Franz

    2017-01-01

    The increasing quality of atmospheric spectroscopy observations has indicated the limitations of the Voigt profile routinely used for line-by-line modeling, and physical processes beyond pressure and Doppler broadening have to be considered. The speed-dependent Voigt (SDV) profile can be readily computed as the difference of the real part of two complex error functions (i.e. Voigt functions). Using a highly accurate code as a reference, various implementations of the SDV function based on Humlíček's rational approximations are examined for typical speed dependences of pressure broadening and the range of wavenumber distances and Lorentz to Doppler width ratios encountered in infrared applications. Neither of these implementations appears to be optimal, and a new algorithm based on a combination of the Humlíček (1982) and Weideman (1994) rational approximations is suggested.

  8. Energy dependence of hadronic activity

    NASA Astrophysics Data System (ADS)

    Gabriel, T. A.; Groom, D. E.; Job, P. K.; Mokhov, N. V.; Stevenson, G. R.

    1994-01-01

    Two features of high-energy hadronic cascades have long been known to shielding specialists: a) in a high-energy hadronic cascade in a given material (incident E ≳ 10 GeV), the relative abundance and spectrum of each hadronic species responsible for most of the energy deposition is independent of the energy or species of the incident hadron, and b) because π0 production bleeds off more and more energy into the electromagnetic sector as the energy of the incident hadron increases, the absolute level of this low-energy hadronic activity ( E ≲ 1 GeV) rises less rapidly than the incident energy, and in fact rises very nearly as a power of the incident energy. Both features are of great importance in hadron calorimetry, where it is the "universal spectrum" which makes possible the definition of an intrinsic {e}/{h}, and the increasing fraction of the energy going into π0's which leads to the energy dependence of {e}/{π}. We present evidence for the "universal spectrum," and use an induction argument and simulation results to demonstrate that the low-energy activity ss Em, with 0.80 ≲ m ≲ 0.85. The hadronic activity produced by incident pions is 15-20% less than that initiated by protons.

  9. Wilderness Adventure Programs: An Activity Profile.

    ERIC Educational Resources Information Center

    Lowenstein, Daniel H.

    Focusing upon the adventure aspect of wilderness programs, this paper presents a profile of those program activities which create a number of challenges and often stressful situations as the means of attaining specified goals and which can best be incorporated under the term "Wilderness Adventure Program" (WAP). Providing information of…

  10. Pathway Activity Profiling (PAPi): from the metabolite profile to the metabolic pathway activity.

    PubMed

    Aggio, Raphael B M; Ruggiero, Katya; Villas-Bôas, Silas Granato

    2010-12-01

    Metabolomics is one of the most recent omics-technologies and uses robust analytical techniques to screen low molecular mass metabolites in biological samples. It has evolved very quickly during the last decade. However, metabolomics datasets are considered highly complex when used to relate metabolite levels to metabolic pathway activity. Despite recent developments in bioinformatics, which have improved the quality of metabolomics data, there is still no straightforward method capable of correlating metabolite level to the activity of different metabolic pathways operating within the cells. Thus, this kind of analysis still depends on extremely laborious and time-consuming processes. Here, we present a new algorithm Pathway Activity Profiling (PAPi) with which we are able to compare metabolic pathway activities from metabolite profiles. The applicability and potential of PAPi was demonstrated using a previously published data from the yeast Saccharomyces cerevisiae. PAPi was able to support the biological interpretations of the previously published observations and, in addition, generated new hypotheses in a straightforward manner. However, PAPi is time consuming to perform manually. Thus, we also present here a new R-software package (PAPi) which implements the PAPi algorithm and facilitates its usage to quickly compare metabolic pathways activities between different experimental conditions. Using the identified metabolites and their respective abundances as input, the PAPi package calculates pathways' Activity Scores, which represents the potential metabolic pathways activities and allows their comparison between conditions. PAPi also performs principal components analysis and analysis of variance or t-test to investigate differences in activity level between experimental conditions. In addition, PAPi generates comparative graphs highlighting up- and down-regulated pathway activity. These datasets are available in http://www.4shared

  11. Abnormal amygdala activation profile in pedophilia.

    PubMed

    Sartorius, Alexander; Ruf, Matthias; Kief, Christine; Demirakca, Traute; Bailer, Josef; Ende, Gabriele; Henn, Fritz A; Meyer-Lindenberg, Andreas; Dressing, Harald

    2008-08-01

    Despite considerable public interest research in neurobiological correlates of pedophilia is scarce. Since amygdala activation is central for emotional valuation, arousal, and salience, we investigated the activation profile of this structure in 10 male subjects with pedophilia (exclusively attracted to boys), all convicted sex-offenders and sentenced to forensic psychiatric treatment along with ten male heterosexual matched controls. We used a sexually non-explicit functional Magnetic Resonance Imaging (fMRI) paradigm with images of men, women, boys or girls randomly embedded in neutral target/non-target geometrical symbols. We applied statistical parametric mapping (SPM2) and SPSS 14 for image processing and analysis. While controls activated significantly less to pictures of children compared to adults, the activation profile was reversed in subjects with pedophilia, who exhibited significantly more activation to children than adults. The highest activation was observed for boys in the patient group, and for women in control participants. Our data show enhanced activation to children's pictures even in an incidental context and suggest the provocative hypothesis that a normally present mechanism for reduced emotional arousal for children relative to adults is reversed in pedophilia, suggesting a neural substrate associated with deviant sexual preference in this condition. More extensive research in this field would be of benefit for both the victims and the offenders.

  12. Quantitative Structure-Activity Relationship Modeling of Kinase Selectivity Profiles.

    PubMed

    Kothiwale, Sandeepkumar; Borza, Corina; Pozzi, Ambra; Meiler, Jens

    2017-09-19

    The discovery of selective inhibitors of biological target proteins is the primary goal of many drug discovery campaigns. However, this goal has proven elusive, especially for inhibitors targeting the well-conserved orthosteric adenosine triphosphate (ATP) binding pocket of kinase enzymes. The human kinome is large and it is rather difficult to profile early lead compounds against around 500 targets to gain an upfront knowledge on selectivity. Further, selectivity can change drastically during derivatization of an initial lead compound. Here, we have introduced a computational model to support the profiling of compounds early in the drug discovery pipeline. On the basis of the extensive profiled activity of 70 kinase inhibitors against 379 kinases, including 81 tyrosine kinases, we developed a quantitative structure-activity relation (QSAR) model using artificial neural networks, to predict the activity of these kinase inhibitors against the panel of 379 kinases. The model's performance in predicting activity ranges from 0.6 to 0.8 depending on the kinase, from the area under the curve (AUC) of the receiver operating characteristics (ROC). The profiler is available online at http://www.meilerlab.org/index.php/servers/show?s_id=23.

  13. Line profile asymmetries in chromospherically active stars

    NASA Technical Reports Server (NTRS)

    Dempsey, Robert C.; Bopp, Bernard W.; Strassmeier, Klaus G.; Granados, Arno F.; Henry, Gregory W.; Hall, Douglas S.

    1992-01-01

    A powerful, new probe of chromospheric activity, cross-correlation, has been developed and applied to a variety of stars. In this particular application, an entire CCD spectrum of an active star is correlated with the spectrum of a narrow-line, inactive star of similar spectral type and luminosity class. Using a number of strong lines in this manner enables the detection of absorption profile asymmetries at moderate resolution (lambda/Delta lambda about 40,000) and S/N 150:1. This technique has been applied to 14 systems mostly RS CVn's, with 10 not greater than nu sin i not greater than 50 km/s and P not less than 7 d. Distortions were detected for the first time in five systems: Sigma Gem, IM Peg, GX Lib, UV Crb, and Zeta And. Detailed modeling, incorporating both spectral line profiles and broad-band photometry, is applied to Sigma Gem. Profile asymmetries for this star are fitted by two high-latitude spots covering 5 percent of the stellar surface. The derived spot temperature of 3400 K is lower than found in previous studies. In addition, two well-known systems have been studied: HD 199178 and V711 Tau. Polar spots are found on both.

  14. An Investigation of Activity Profiles of Older Adults

    PubMed Central

    Putnam, Michelle; Lee, Yung Soo; Greenfield, Jennifer C.; Inoue, Megumi; Chen, Huajuan

    2014-01-01

    Objectives. In this study, we advance knowledge about activity engagement by considering many activities simultaneously to identify profiles of activity among older adults. Further, we use cross-sectional data to explore factors associated with activity profiles and prospective data to explore activity profiles and well-being outcomes. Method. We used the core survey data from the years 2008 and 2010, as well as the 2009 Health and Retirement Study Consumption and Activities Mail Survey (HRS CAMS). The HRS CAMS includes information on types and amounts of activities. We used factor analysis and latent class analysis to identify activity profiles and regression analyses to assess antecedents and outcomes associated with activity profiles. Results. We identified 5 activity profiles: Low Activity, Moderate Activity, High Activity, Working, and Physically Active. These profiles varied in amount and type of activities. Demographic and health factors were related to profiles. Activity profiles were subsequently associated with self-rated health and depression symptoms. Discussion. The use of a 5-level categorical activity profile variable may allow more complex analyses of activity that capture the “whole person.” There is clearly a vulnerable group of low-activity individuals as well as a High Activity group that may represent the “active ageing” vision. PMID:24526690

  15. Strain-dependent profile of misfolded prion protein aggregates

    PubMed Central

    Morales, Rodrigo; Hu, Ping Ping; Duran-Aniotz, Claudia; Moda, Fabio; Diaz-Espinoza, Rodrigo; Chen, Baian; Bravo-Alegria, Javiera; Makarava, Natallia; Baskakov, Ilia V.; Soto, Claudio

    2016-01-01

    Prions are composed of the misfolded prion protein (PrPSc) organized in a variety of aggregates. An important question in the prion field has been to determine the identity of functional PrPSc aggregates. In this study, we used equilibrium sedimentation in sucrose density gradients to separate PrPSc aggregates from three hamster prion strains (Hyper, Drowsy, SSLOW) subjected to minimal manipulations. We show that PrPSc aggregates distribute in a wide range of arrangements and the relative proportion of each species depends on the prion strain. We observed a direct correlation between the density of the predominant PrPSc aggregates and the incubation periods for the strains studied. The relative presence of PrPSc in fractions of different sucrose densities was indicative of the protein deposits present in the brain as analyzed by histology. Interestingly, no association was found between sensitivity to proteolytic degradation and aggregation profiles. Therefore, the organization of PrP molecules in terms of the density of aggregates generated may determine some of the particular strain properties, whereas others are independent from it. Our findings may contribute to understand the mechanisms of strain variation and the role of PrPSc aggregates in prion-induced neurodegeneration. PMID:26877167

  16. Strain-dependent profile of misfolded prion protein aggregates.

    PubMed

    Morales, Rodrigo; Hu, Ping Ping; Duran-Aniotz, Claudia; Moda, Fabio; Diaz-Espinoza, Rodrigo; Chen, Baian; Bravo-Alegria, Javiera; Makarava, Natallia; Baskakov, Ilia V; Soto, Claudio

    2016-02-15

    Prions are composed of the misfolded prion protein (PrP(Sc)) organized in a variety of aggregates. An important question in the prion field has been to determine the identity of functional PrP(Sc) aggregates. In this study, we used equilibrium sedimentation in sucrose density gradients to separate PrP(Sc) aggregates from three hamster prion strains (Hyper, Drowsy, SSLOW) subjected to minimal manipulations. We show that PrP(Sc) aggregates distribute in a wide range of arrangements and the relative proportion of each species depends on the prion strain. We observed a direct correlation between the density of the predominant PrP(Sc) aggregates and the incubation periods for the strains studied. The relative presence of PrP(Sc) in fractions of different sucrose densities was indicative of the protein deposits present in the brain as analyzed by histology. Interestingly, no association was found between sensitivity to proteolytic degradation and aggregation profiles. Therefore, the organization of PrP molecules in terms of the density of aggregates generated may determine some of the particular strain properties, whereas others are independent from it. Our findings may contribute to understand the mechanisms of strain variation and the role of PrP(Sc) aggregates in prion-induced neurodegeneration.

  17. Activity-Based Protein Profiling of Microbes

    SciTech Connect

    Sadler, Natalie C.; Wright, Aaron T.

    2015-02-01

    Activity-Based Protein Profiling (ABPP) in conjunction with multimodal characterization techniques has yielded impactful findings in microbiology, particularly in pathogen, bioenergy, drug discovery, and environmental research. Using small molecule chemical probes that react irreversibly with specific proteins or protein families in complex systems has provided insights in enzyme functions in central metabolic pathways, drug-protein interactions, and regulatory protein redox, for systems ranging from photoautotrophic cyanobacteria to mycobacteria, and combining live cell or cell extract ABPP with proteomics, molecular biology, modeling, and other techniques has greatly expanded our understanding of these systems. New opportunities for application of ABPP to microbial systems include: enhancing protein annotation, characterizing protein activities in myriad environments, and reveal signal transduction and regulatory mechanisms in microbial systems.

  18. Salivary lysosomal exoglycosidases profiles in patients with insulin-dependent and noninsulin-dependent diabetes mellitus.

    PubMed

    Zalewska, Anna; Knaś, Małgorzata; Niczyporuk, Marek; Razak, Hady Hady; Waszkiewicz, Napoleon; Przystupa, Adrian Wojciech; Waszkiel, Danuta; Zarzycki, Wiesław

    2013-01-01

    In this study we have investigated the effects of type I (insulin-dependent) and II (non-insulin dependent) diabetes mellitus on the specific activity and the output of salivary exoglycosidases: N-acetyl-β-hexosoaminidase (HEX), and its isoenzymes A and B (HEX A, HEX B), and β-glucuronidase (GLU) in well controlled diabetic patients compared to healthy age-matched controls. In the saliva HEX, HEX A, HEX B and GLU were determined according to Marciniak et al. Protein was determined by the Lowry et al. method. Our results show that in the case of type I diabetes, the significantly increased activity of salivary total HEX is mainly due to the significantly increased HEX A specific activity. Significantly increased HEX specific activity in DM II is an outcome of significantly increased HEX A as well as HEX B specific activities in comparison to the appropriate healthy control. Our results showed a significant increase in the specific activity of GLU in saliva of type II diabetes patients. The output of lysosomal exoglycosidases showed a similar significant increase compared to the healthy control, in both groups of diabetes mellitus patients. This study has demonstrated that non-insulin dependent diabetes mellitus more strongly modify salivary glands glycoconjugates catabolism, which can be attributed to functional and morphological changes. A significant increase in the outputs of exoglycosidases in saliva of both type diabetes patients once more indicates that special attention should be paid to the oral health of these patients.

  19. Passion and dependency in online shopping activities.

    PubMed

    Wang, Chih-Chien; Yang, Hui-Wen

    2007-04-01

    This study examines the influence of harmonious passion (HP) and obsessive passion (OP) to online shopping dependency. The results show that both HP and OP might lead to online shopping dependency and online shoppers with OP are more dependent on online shopping activities. In addition, this study also found out that HP and OP could be denoted as a sequence of different intensities of passion, where HP might be a necessity of OP.

  20. Cohort Profile: The Nicotine Dependence in Teens (NDIT) Study.

    PubMed

    O'Loughlin, Jennifer; Dugas, Erika N; Brunet, Jennifer; DiFranza, Joseph; Engert, James C; Gervais, Andre; Gray-Donald, Katherine; Karp, Igor; Low, Nancy C; Sabiston, Catherine; Sylvestre, Marie-Pierre; Tyndale, Rachel F; Auger, Nathalie; Auger, Nathalie; Mathieu, Belanger; Tracie, Barnett; Chaiton, Michael; Chenoweth, Meghan J; Constantin, Evelyn; Contreras, Gisèle; Kakinami, Lisa; Labbe, Aurelie; Maximova, Katerina; McMillan, Elizabeth; O'Loughlin, Erin K; Pabayo, Roman; Roy-Gagnon, Marie-Hélène; Tremblay, Michèle; Wellman, Robert J; Hulst, Andraeavan; Paradis, Gilles

    2015-10-01

    The Nicotine Dependence in Teens (NDIT) study is a prospective cohort investigation of 1294 students recruited in 1999-2000 from all grade 7 classes in a convenience sample of 10 high schools in Montreal, Canada. Its primary objectives were to study the natural course and determinants of cigarette smoking and nicotine dependence in novice smokers. The main source of data was self-report questionnaires administered in class at school every 3 months from grade 7 to grade 11 (1999-2005), for a total of 20 survey cycles during high school education. Questionnaires were also completed after graduation from high school in 2007-08 and 2011-12 (survey cycles 21 and 22, respectively) when participants were aged 20 and 24 years on average, respectively. In addition to its primary objectives, NDIT has embedded studies on obesity, blood pressure, physical activity, team sports, sedentary behaviour, diet, genetics, alcohol use, use of illicit drugs, second-hand smoke, gambling, sleep and mental health. Results to date are described in 58 publications, 20 manuscripts in preparation, 13 MSc and PhD theses and 111 conference presentations. Access to NDIT data is open to university-appointed or affiliated investigators and to masters, doctoral and postdoctoral students, through their primary supervisor (www.nditstudy.ca).

  1. Cohort Profile: The Nicotine Dependence in Teens (NDIT) Study

    PubMed Central

    O’Loughlin, Jennifer; Dugas, Erika N; Brunet, Jennifer; DiFranza, Joseph; Engert, James C; Gervais, Andre; Gray-Donald, Katherine; Karp, Igor; Low, Nancy C; Sabiston, Catherine; Sylvestre, Marie-Pierre; Tyndale, Rachel F; Auger, Nathalie; Barnett, Tracie; Mathieu, Bélanger; Chaiton, Michael; Chenoweth, Meghan J; Constantin, Evelyn; Contreras, Gisèle; Kakinami, Lisa; Labbe, Aurélie; Maximova, Katerina; McMillan, Elizabeth; O’Loughlin, Erin K; Pabayo, Roman; Roy-Gagnon, Marie-Hélène; Tremblay, Michèle; Wellman, Robert J; van Hulst, Andraea; Paradis, Gilles

    2015-01-01

    The Nicotine Dependence in Teens (NDIT) study is a prospective cohort investigation of 1294 students recruited in 1999–2000 from all grade 7 classes in a convenience sample of 10 high schools in Montreal, Canada. Its primary objectives were to study the natural course and determinants of cigarette smoking and nicotine dependence in novice smokers. The main source of data was self-report questionnaires administered in class at school every 3 months from grade 7 to grade 11 (1999–2005), for a total of 20 survey cycles during high school education. Questionnaires were also completed after graduation from high school in 2007–08 and 2011–12 (survey cycles 21 and 22, respectively) when participants were aged 20 and 24 years on average, respectively. In addition to its primary objectives, NDIT has embedded studies on obesity, blood pressure, physical activity, team sports, sedentary behaviour, diet, genetics, alcohol use, use of illicit drugs, second-hand smoke, gambling, sleep and mental health. Results to date are described in 58 publications, 20 manuscripts in preparation, 13 MSc and PhD theses and 111 conference presentations. Access to NDIT data is open to university-appointed or affiliated investigators and to masters, doctoral and postdoctoral students, through their primary supervisor (www.nditstudy.ca). PMID:25022274

  2. Activity dependent CAM cleavage and neurotransmission

    PubMed Central

    Conant, Katherine; Allen, Megan; Lim, Seung T.

    2015-01-01

    Spatially localized proteolysis represents an elegant means by which neuronal activity dependent changes in synaptic structure, and thus experience dependent learning and memory, can be achieved. In vitro and in vivo studies suggest that matrix metalloproteinase and adamalysin activity is concentrated at the cell surface, and emerging evidence suggests that increased peri-synaptic expression, release and/or activation of these proteinases occurs with enhanced excitatory neurotransmission. Synaptically expressed cell adhesion molecules (CAMs) could therefore represent important targets for neuronal activity-dependent proteolysis. Several CAM subtypes are expressed at the synapse, and their cleavage can influence the efficacy of synaptic transmission through a variety of non-mutually exclusive mechanisms. In the following review, we discuss mechanisms that regulate neuronal activity-dependent synaptic CAM shedding, including those that may be calcium dependent. We also highlight CAM targets of activity-dependent proteolysis including neuroligin and intercellular adhesion molecule-5 (ICAM-5). We include discussion focused on potential consequences of synaptic CAM shedding, with an emphasis on interactions between soluble CAM cleavage products and specific pre- and post-synaptic receptors. PMID:26321910

  3. Strength Profiles of the Continental Lithosphere: Fabric Dependence, Strain Dependence, and Implications for Stability and Localization

    NASA Astrophysics Data System (ADS)

    Montesi, L.; Gueydan, F.

    2014-12-01

    A fundamental characteristic of plate tectonics is the contrast between strong plate interiors and weak plate boundaries. The weakness has both a structural aspect, as deformation zones can be reactivated during different tectonic episodes, and a dynamic aspect, as there is no major compositional difference between the lithosphere at active and inactive regions. These characteristics point to a structural origin of the weakness of plate boundaries, as deformation affect structures but structures do not heal rapidly. This presentation summarizes recent advances made in estimating the rheology of polymineralic aggregates under conditions appropriate for various levels of the lithosphere. We construct a new generation of strength profiles that take into consideration the differences in microstructure that are observed in continental shear zones: reduced grain size, metamorphism, and weak phase interconnection/layering. For each lithosphere stratigraphy, two strength profiles can be constructed, one for the undeformed material, the other for a high-strain material. The low-strain profile assumes a reference strain rate, constant with depth, an untextured mixture of plagioclase and biotite in the upper and middle crust (biotite being unstable in the lower crust, and an olivine-pyroxene untextured mixture in the mantle. As deformation proceeds, the fabric evolves until the phases become interconnected, and olivine grain size is reduced in the upper mantle. A high-strain strength profile can be derived for constant strain rate for reference. However, strain rate is likely higher in the high-strain state, but the degree of this enhancement is poorly constrained. We can follow two hypotheses. In the first, stress at each depth is the same as in the initial state. The strength profile is unmodified, but we can solve at each depth for a new strain rate. Alternatively, we can impose that the stress integrated over a lithospheric column is the same and solve for a depth

  4. Depth and size dependence of Mn-53 activity in chondrites

    NASA Astrophysics Data System (ADS)

    Bhattacharya, S. K.; Imamura, M.; Sinha, N.; Bhandari, N.

    1980-11-01

    The dependence of Mn-53 activity in core samples from four chondrites on sample shielding depth and the pre-atmospheric size of the meteorite is investigated. Cosmogenic Mn-53 activity and cosmic ray track densities were measured at various depths in cores from the Madhipura, Udaipur, Bansur and St. Severin chondrites. Track density analyses indicate effective radii of 6.5, 9, 15 and 25 cm for the meteorites in space, respectively. The depth profiles of Mn-53 activity reveal that the nuclide production rate at any given depth increases with meteorite size, while the activity profile for meteorites with effective radii less than or equal to 15 cm is nearly flat for shielding depths greater than 3 cm and that for meteorites about 25 cm in radius increases by about 40% from near the surface to the center, indicating the importance of the secondary cascade at radii greater than 15 cm. The profiles are then used to determine the spectral hardness parameter as a function of depth which is in turn used in the calculation of production profiles for Al-26 which are found to be in agreement with observations.

  5. Activity profile in elite Italian soccer team.

    PubMed

    Vigne, G; Gaudino, C; Rogowski, I; Alloatti, G; Hautier, C

    2010-05-01

    The purpose of this study was to analyse the activity profile of players in a top-class team in the Italian national football league over the course of a season (n=388). The effect of playing position and the two halves on the number and duration of short, intense bursts of effort and recovery phases was studied. The main results show that midfielders cover significantly more distance than players in other positions (p<0.001). For midfielders, the number of displacements of 2-40 m and the number of sprints covering between 2 and 9 m and between 30 and 40 m are considerably greater than for other positions (p<0.05). The distances covered in the second half compared to the first half are significantly lower for all categories of run (p<0.05). In the second half, the distance covered at very high intensity is significantly lower (p<0.01), whilst the number of recovery times greater than 120 s increases significantly compared to the first half (p<0.01). This study provides data which could be used as a basis for the work of scientists as well as football professionals.

  6. The safety profile of drotrecogin alfa (activated)

    PubMed Central

    Fumagalli, Roberto; Mignini, Mariano A

    2007-01-01

    Continued safety assessment is essential for any newly approved therapy. Drotrecogin alfa (activated; DrotAA), which is approved for use in severe sepsis, has undergone clinical trials with corresponding safety analyses since 1995. However, the only comprehensive review of all trials is that reported in 2003 by Bernard and coworkers. This is an important review that focuses on the safety profile of DrotAA in all published studies (six randomized clinical trials and five national registry studies) and in previously unpublished data. DrotAA treatment is associated with an increased risk for bleeding (which in general is manageable). Nevertheless, the available evidence shows that any adverse effects of DrotAA are outweighed by its benefits in patients with severe sepsis who are at high risk for death. So far, more than 9,000 patients have been enrolled in clinical trials involving DrotAA, providing a valuable safety database. It is of note that although DrotAA does increase the risk of bleeding, this has not been associated with an overall increase in the rate of all severe adverse events. PMID:18269693

  7. Interrelationship of antioxidative status, lipid peroxidation, and lipid profile in insulin-dependent and non-insulin-dependent diabetic patients.

    PubMed

    Cimbaljević, Branko; Vasilijević, Ana; Cimbaljević, Slavica; Buzadzić, Biljana; Korać, Aleksandra; Petrović, Vesna; Janković, Aleksandra; Korać, Bato

    2007-10-01

    This study aimed to investigate the interrelationship of plasma lipid profile, lipid peroxidation, and erythrocyte antioxidative defense in patients with insulin-dependent (IDDM) and non-insulin-dependent (NIDDM) diabetes mellitus. Plasma levels of total cholesterol, triglycerides, and lipid peroxides and the activities of copper, zinc superoxide dismutase (CuZnSOD), catalase, glutathione peroxidase (GSH-Px), as well as the amount of glutathione in erythrocytes, were determined in IDDM, NIDDM, and nondiabetic control subjects. Additionally, morphology of erythrocytes in all subjects was examined. Plasma levels of total cholesterol and triglycerides were significantly increased in NIDDM compared with controls. Also, the lipid peroxide level was higher in NIDDM than in either control or IDDM subjects. CuZnSOD activity in erythrocytes was elevated in NIDDM patients compared with the control. In NIDDM patients, more extensive erythrocyte spherocytosis and echinocytosis compared with both control and IDDM subjects were observed. In contrast with the IDDM group, the observed abnormality in lipid metabolism in NIDDM patients is closely associated with increased lipid peroxidation, changes in antioxidative defense, and erythrocyte morphology.

  8. Global Profiling of Carbohydrate Active Enzymes in Human Gut Microbiome

    PubMed Central

    Mande, Sharmila S.

    2015-01-01

    Motivation Carbohydrate Active enzyme (CAZyme) families, encoded by human gut microflora, play a crucial role in breakdown of complex dietary carbohydrates into components that can be absorbed by our intestinal epithelium. Since nutritional wellbeing of an individual is dependent on the nutrient harvesting capability of the gut microbiome, it is important to understand how CAZyme repertoire in the gut is influenced by factors like age, geography and food habits. Results This study reports a comprehensive in-silico analysis of CAZyme profiles in the gut microbiomes of 448 individuals belonging to different geographies, using similarity searches of the corresponding gut metagenomic contigs against the carbohydrate active enzymes database. The study identifies a core group of 89 CAZyme families that are present across 85% of the gut microbiomes. The study detects several geography/age-specific trends in gut CAZyme repertoires of the individuals. Notably, a group of CAZymes having a positive correlation with BMI has been identified. Further this group of BMI-associated CAZymes is observed to be specifically abundant in the Firmicutes phyla. One of the major findings from this study is identification of three distinct groups of individuals, referred to as 'CAZotypes', having similar CAZyme profiles. Distinct taxonomic drivers for these CAZotypes as well as the probable dietary basis for such trends have also been elucidated. The results of this study provide a global view of CAZyme profiles across individuals of various geographies and age-groups. These results re-iterate the need of a more precise understanding of the role of carbohydrate active enzymes in human nutrition. PMID:26544883

  9. Neurobiological Signatures of Alcohol Dependence Revealed by Protein Profiling

    PubMed Central

    Gorini, Giorgio; Roberts, Amanda J.; Mayfield, R. Dayne

    2013-01-01

    Alcohol abuse causes dramatic neuroadaptations in the brain, which contribute to tolerance, dependence, and behavioral modifications. Previous proteomic studies in human alcoholics and animal models have identified candidate alcoholism-related proteins. However, recent evidences suggest that alcohol dependence is caused by changes in co-regulation that are invisible to single protein-based analysis. Here, we analyze global proteomics data to integrate differential expression, co-expression networks, and gene annotations to unveil key neurobiological rearrangements associated with the transition to alcohol dependence modeled by a Chronic Intermittent Ethanol (CIE), two-bottle choice (2BC) paradigm. We analyzed cerebral cortices (CTX) and midbrains (MB) from male C57BL/6J mice subjected to a CIE, 2BC paradigm, which induces heavy drinking and represents one of the best available animal models for alcohol dependence and relapse drinking. CIE induced significant changes in protein levels in dependent mice compared with their non-dependent controls. Multiple protein isoforms showed region-specific differential regulation as a result of post-translational modifications. Our integrative analysis identified modules of co-expressed proteins that were highly correlated with CIE treatment. We found that modules most related to the effects of CIE treatment coordinate molecular imbalances in endocytic- and energy-related pathways, with specific proteins involved, such as dynamin-1. The qRT-PCR experiments validated both differential and co-expression analyses, and the correspondence among our data and previous genomic and proteomic studies in humans and rodents substantiates our findings. The changes identified above may play a key role in the escalation of ethanol consumption associated with dependence. Our approach to alcohol addiction will advance knowledge of brain remodeling mechanisms and adaptive changes in response to drug abuse, contribute to understanding of

  10. NMDA receptor-dependent synaptic plasticity in dorsal and intermediate hippocampus exhibits distinct frequency-dependent profiles.

    PubMed

    Kenney, Jana; Manahan-Vaughan, Denise

    2013-11-01

    The hippocampus may be functionally differentiated along its dorsoventral axis. In contrast to the wealth of data available on synaptic plasticity mechanisms in the dorsal hippocampus, little is known about synaptic plasticity processes in the intermediate hippocampus. Behavioral data suggest that this structure may play a distinct role in learning and memory. Here, we compared amplitudes, frequency-dependency and persistency of long-term potentiation (LTP) and long-term depression (LTD) in the dorsal (DDG) and intermediate dentate gyrus (IDG). In freely moving rats, high-frequency stimulation (HFS) at 200 Hz (10 burst of 15 stimuli) elicited LTP of similar magnitude in both structures that persisted for over 24 h. The intermediate dentate gyrus is more likely to exhibit persistent LTP than its dorsal counterpart, however: HFS at 200 Hz (3 or 1 burst(s)) or 100 Hz elicited short-term potentiation (STP) in DDG, unlike in the IDG, where LTP could be recorded for at least 4 h. Whereas low frequency stimulation (LFS) at 1 Hz elicited long-lasting LTD (>24 h) in the DDG, it had no significant effect on fEPSP profile in the IDG. LFS at 2 Hz elicited short-term depression in DDG and had no effect in IDG. LTP in both IDG and DDG required activation of N-methyl-D-aspartate receptors. Paired-pulse and input-output responses differed in IDG and DDG. Our data suggest that afferent input from the entorhinal cortex generates a different response profile in the dorsal vs. intermediate DG, which may in turn relate to their postulated distinct roles in synaptic information processing and memory formation. This article is part of the Special Issue entitled 'Glutamate Receptor-Dependent Synaptic Plasticity'. Copyright © 2013 Elsevier Ltd. All rights reserved.

  11. Chemical transformations that yield compounds with distinct activity profiles.

    PubMed

    Hu, Ye; Bajorath, Jürgen

    2011-07-14

    We have systematically searched for chemical changes that generate compounds with distinct biological activity profiles. For this purpose, activity profiles were generated for ∼42000 compounds active against human targets. Unique activity profiles involving multiple target proteins were determined, and all possible matched molecular pairs (MMPs) were identified for compounds representing these profiles. An MMP is defined as a pair of compounds that are distinguished from each other only at a single site such as an R group or ring system. For example, in an MMP, a hydroxyl group might be replaced by a halogen atom or a benzene ring by an amide group. From ∼37500 MMPs, more than 300 nonredundant chemical transformations were isolated that yielded compounds with distinct activity profiles. None of these transformations was found in pairs of compounds with overlapping activity profiles. These transformations were ranked according to the number of MMPs, the number of activity profiles, and the total number of targets that they covered. In many instances, prioritized transformations involved ring systems of varying complexity. All transformations that were found to switch activity profiles are provided to enable further analysis and aid in compound design efforts.

  12. Plasma Metabolic Profiles in Women are Menopause Dependent.

    PubMed

    Ke, Chaofu; Hou, Yan; Zhang, Haiyu; Yang, Kai; Wang, Jingtao; Guo, Bing; Zhang, Fan; Li, Hailong; Zhou, Xiaohua; Li, Ying; Li, Kang

    2015-01-01

    Menopause is an endocrinological transition that greatly affects health and disease susceptibility in middle-aged and elderly women. To gain new insights into the metabolic process of menopause, plasma metabolic profiles in 115 pre- and post-menopausal women were systematically analyzed by ultra-performance liquid chromatography/mass spectrometry in conjunction with univariate and multivariate statistical analysis. Metabolic signatures revealed considerable differences between pre- and post-menopausal women, and clear separations were observed between the groups in partial least-squares discriminant analysis score plots. In total, 28 metabolites were identified as potential metabolite markers for menopause, including up-regulated acylcarnitines, fatty acids, lysophosphatidylcholines, lysophosphatidylethanolamines, and down-regulated pregnanediol-3-glucuronide, dehydroepiandrosterone sulfate, p-hydroxyphenylacetic acid and dihydrolipoic acid. These differences highlight that significant alterations occur in fatty acid β-oxidation, phospholipid metabolism, hormone metabolism and amino acid metabolism in post-menopausal women. In conclusion, our plasma metabolomics study provides novel understanding of the metabolic profiles related to menopause, and will be useful for investigating menopause-related diseases and assessing metabolomic confounding factors.

  13. Building on Student Diversity: Profiles and Activities

    ERIC Educational Resources Information Center

    Cowdery, Joy R.; Rogness, Linda Ingling; Morrow, Linda E.; Wilson, Vicki A.

    2006-01-01

    This text captures the profiles and cumulative records of six diverse students at early childhood, middle childhood, and then high school level. Intended for the preservice teacher, this book illustrates how to create a caring school environment; accommodate for special learning needs in instructional and assessments; and interact with families…

  14. Building on Student Diversity: Profiles and Activities

    ERIC Educational Resources Information Center

    Cowdery, Joy R.; Rogness, Linda Ingling; Morrow, Linda E.; Wilson, Vicki A.

    2006-01-01

    This text captures the profiles and cumulative records of six diverse students at early childhood, middle childhood, and then high school level. Intended for the preservice teacher, this book illustrates how to create a caring school environment; accommodate for special learning needs in instructional and assessments; and interact with families…

  15. Light-Dependent Electrogenic Activity of Cyanobacteria

    PubMed Central

    Baskakov, Ilia V.

    2010-01-01

    Background Cyanobacteria account for 20–30% of Earth's primary photosynthetic productivity and convert solar energy into biomass-stored chemical energy at the rate of ∼450 TW [1]. These single-cell microorganisms are resilient predecessors of all higher oxygenic phototrophs and can be found in self-sustaining, nitrogen-fixing communities the world over, from Antarctic glaciers to the Sahara desert [2]. Methodology/Principal Findings Here we show that diverse genera of cyanobacteria including biofilm-forming and pelagic strains have a conserved light-dependent electrogenic activity, i.e. the ability to transfer electrons to their surroundings in response to illumination. Naturally-growing biofilm-forming photosynthetic consortia also displayed light-dependent electrogenic activity, demonstrating that this phenomenon is not limited to individual cultures. Treatment with site-specific inhibitors revealed the electrons originate at the photosynthetic electron transfer chain (P-ETC). Moreover, electrogenic activity was observed upon illumination only with blue or red but not green light confirming that P-ETC is the source of electrons. The yield of electrons harvested by extracellular electron acceptor to photons available for photosynthesis ranged from 0.05% to 0.3%, although the efficiency of electron harvesting likely varies depending on terminal electron acceptor. Conclusions/Significance The current study illustrates that cyanobacterial electrogenic activity is an important microbiological conduit of solar energy into the biosphere. The mechanism responsible for electrogenic activity in cyanobacteria appears to be fundamentally different from the one exploited in previously discovered electrogenic bacteria, such as Geobacter, where electrons are derived from oxidation of organic compounds and transported via a respiratory electron transfer chain (R-ETC) [3], [4]. The electrogenic pathway of cyanobacteria might be exploited to develop light-sensitive devices or

  16. Sample storage for soil enzyme activity and bacterial community profiles.

    PubMed

    Wallenius, K; Rita, H; Simpanen, S; Mikkonen, A; Niemi, R M

    2010-04-01

    Storage of samples is often an unavoidable step in environmental data collection, since available analytical capacity seldom permits immediate processing of large sample sets needed for representative data. In microbiological soil studies, sample pretreatments may have a strong influence on measurement results, and thus careful consideration is required in the selection of storage conditions. The aim of this study was to investigate the suitability of prolonged (up to 16 weeks) frozen or air-dried storage for divergent soil materials. The samples selected to this study were mineral soil (clay loam) from an agricultural field, humus from a pine forest and compost from a municipal sewage sludge composting field. The measured microbiological parameters included functional profiling with ten different hydrolysing enzyme activities determined by artificial fluorogenic substrates, and structural profiling with bacterial 16S rDNA community fingerprints by amplicon length heterogeneity analysis (LH-PCR). Storage of samples affected the observed fluorescence intensity of the enzyme assay's fluorophor standards dissolved in soil suspension. The impact was highly dependent on the soil matrix and storage method, making it important to use separate standardisation for each combination of matrix type, storage method and time. Freezing proved to be a better storage method than air-drying for all the matrices and enzyme activities studied. The effect of freezing on the enzyme activities was small (<20%) in clay loam and forest humus and moderate (generally 20-30%) in compost. The most dramatic decreases (>50%) in activity were observed in compost after air-drying. The bacterial LH-PCR community fingerprints were unaffected by frozen storage in all matrices. The effect of storage treatments was tested using a new statistical method based on showing similarity rather than difference of results.

  17. Neuropsychological profiling of impulsivity and compulsivity in cocaine dependent individuals.

    PubMed

    Fernández-Serrano, María José; Perales, José César; Moreno-López, Laura; Pérez-García, Miguel; Verdejo-García, Antonio

    2012-01-01

    Research on the relative impact of trait impulsivity vs. drug exposure on neuropsychological probes of response inhibition vs. response perseveration has been posited as a valid pathway to explore the transition between impulsivity and compulsivity on psychostimulant dependence. The objectives of this study are to examine performance differences between cocaine-dependent individuals (CDI) and healthy comparison individuals (HCI) on neuropsychological probes of inhibition and perseveration and to examine the predictive impact of trait impulsivity-a proxy of premorbid vulnerability, and severity of cocaine use-a proxy of drug exposure, on CDI's performance. Forty-two CDI and 65 HCI were assessed using the UPPS-P Scale (trait impulsivity), the Stroop and go/no-go (inhibition) and revised-strategy application and probabilistic reversal tests (perseveration). CDI, compared to HCI, have elevated scores on trait impulsivity and perform significantly poorer on inhibition and perseveration, with specific detrimental effects of duration of cocaine use on perseveration. CDI have both inhibition and perseveration deficits; both patterns were broadly indicative of orbitofrontal dysfunction in the context of reinforcement learning. Impulsive personality and cocaine exposure jointly contribute to deficits in response perseveration or compulsivity.

  18. Age and gender dependent profile of food choice.

    PubMed

    Westenhoefer, Joachim

    2005-01-01

    Several studies have described remarkable differences in food choice between men and women. Consistently, women are reported to have higher intakes of fruit and vegetables, higher intakes of dietary fiber and lower intakes of fat. In accordance with such more healthy food choice, women usually attach greater importance to healthy eating. In addition, the motivation of weight control is more prominent in women and they are more likely to diet or restrain their eating behavior. Recently, studies found that health beliefs and weight control motivation may explain up to 50 percent of gender differences in food choice. In addition, less healthy food choice profiles of men may be related to their poorer nutritional knowledge. However, health beliefs, eating attitudes and dieting appear to be phenomena which vary throughout the life span. In growing older, changes in the chemosensory perceptual systems play an important role in food choice. The decline of gustatory and--perhaps even more pronounced--in olfactory function may lead to a decrease of the pleasantness of food, thus limiting the reinforcing properties of food intake which eventually results in a decrease of appetite, often reported in elderly people. In addition, there are some indications that sensory-specific satiety diminishes with age. Sensory-specific satiety is the reduction in the pleasantness of food as it is consumed. This decrease of pleasantness usually motivates the choice of other foods and therefore, a varied diet. Therefore, the decrease of sensory-specific satiety may in part explain the limited variety of the diet sometimes seen in elderly people. However, lifestyle, socio-economic situation and other variables may limit the influence of such physiological changes and help to maintain an adequate food intake despite these age-related processes.

  19. Dose-dependent effects of alcohol administration on behavioral profiles in the MCSF test.

    PubMed

    Karlsson, Oskar; Roman, Erika

    2016-02-01

    The acute effects of alcohol administration are age-, dose-, time- and task-dependent. Although generally considered to be a sedative drug, alcohol has both stimulatory and depressant effects on behavior, depending on dose and time. Alcohol-induced motor activating effects are consistently shown in mice but rarely demonstrated in adult, outbred rats using conventional behavioral tests. The aim of the present experiment was to study acute alcohol-induced effects on behavioral profiles in a more complex environment using the novel multivariate concentric square field™ (MCSF) test, designed for assessing different behaviors in the same trial including locomotor activity. Adult male Wistar rats (Sca:WI) were administered one intraperitoneal (i.p.) injection of alcohol (0.0 g/kg, 0.5 g/kg, 1.0 g/kg, or 1.5 g/kg) 5 min prior to the 30-min MCSF test. The two highest doses induced marked motor-suppressing effects. A significant interaction between group and time was found in general activity when comparing rats exposed to alcohol at 0.0 g/kg and 0.5 g/kg. In contrast to the 0.0 g/kg dose that increased the activity over time, animals administered the low dose (0.5 g/kg) demonstrated an initial high activity followed by a decline over time. No indications for acute alcohol-induced anxiolytic-like effects were found. The multivariate setting in the MCSF test appears to be sensitive for detecting motor-activating effects of low doses of alcohol as well as reduced locomotion at doses lower than in other behavioral tasks. The detection of subtle changes in behavior across time and dose is important for understanding alcohol-induced effects. This approach may be useful in evaluating alcohol doses that correspond to different degrees of intoxication in humans. Copyright © 2016 The Authors. Published by Elsevier Inc. All rights reserved.

  20. Emerging Drug Profile: Cyclin-Dependent Kinase Inhibitors

    PubMed Central

    Blachly, James S.; Byrd, John C.

    2013-01-01

    As the rational application of targeted therapies in cancer supplants traditional cytotoxic chemotherapy, there is an ever-greater need for a thorough understanding of the complex machinery of the cell and an application of this knowledge to the development of novel therapeutics and combinations of agents. Here, we review the current state of knowledge of the class of targeted agents known as cyclin-dependent kinase (CDK) inhibitors, with a focus on chronic lymphocytic leukemia (CLL). Flavopiridol (alvocidib) is the best studied of the CDK inhibitors, producing a dramatic cytotoxic effect in vitro and in vivo, with the principal limiting factor of acute tumor lysis. Unfortunately, flavopiridol has a narrow therapeutic window and is relatively non-selective with several off-target (i.e. non-CDK) effects, which prompted development of the second-generation CDK inhibitor dinaciclib. Dinaciclib appears to be both more potent and selective than flavopiridol, with at least an order of magnitude greater therapeutic index, and is currently in phase III clinical trials. In additional to flavopiridol and dinaciclib, we also review the current state of other members of this class, and provide commentary as to the future direction of combination therapy including CDK inhibitors. PMID:23488658

  1. Frequency-dependent oscillatory neural profiles during imitation

    PubMed Central

    Sugata, Hisato; Hirata, Masayuki; Tamura, Yuichi; Onishi, Hisao; Goto, Tetsu; Araki, Toshihiko; Yorifuji, Shiro

    2017-01-01

    Imitation is a complex process that includes higher-order cognitive and motor function. This process requires an observation-execution matching system that transforms an observed action into an identical movement. Although the low-gamma band is thought to reflect higher cognitive processes, no studies have focused on it. Here, we used magnetoencephalography (MEG) to examine the neural oscillatory changes including the low-gamma band during imitation. Twelve healthy, right-handed participants performed a finger task consisting of four conditions (imitation, execution, observation, and rest). During the imitation and execution conditions, significant event-related desynchronizations (ERDs) were observed at the left frontal, central, and parietal MEG sensors in the alpha, beta, and low-gamma bands. Functional connectivity analysis at the sensor level revealed an imitation-related connectivity between a group of frontal sensors and a group of parietal sensors in the low-gamma band. Furthermore, source reconstruction with synthetic aperture magnetometry showed significant ERDs in the low-gamma band in the left sensorimotor area and the middle frontal gyrus (MFG) during the imitation condition when compared with the other three conditions. Our results suggest that the oscillatory neural activities of the low-gamma band at the sensorimotor area and MFG play an important role in the observation-execution matching system related to imitation. PMID:28393878

  2. Frequency-dependent oscillatory neural profiles during imitation.

    PubMed

    Sugata, Hisato; Hirata, Masayuki; Tamura, Yuichi; Onishi, Hisao; Goto, Tetsu; Araki, Toshihiko; Yorifuji, Shiro

    2017-04-10

    Imitation is a complex process that includes higher-order cognitive and motor function. This process requires an observation-execution matching system that transforms an observed action into an identical movement. Although the low-gamma band is thought to reflect higher cognitive processes, no studies have focused on it. Here, we used magnetoencephalography (MEG) to examine the neural oscillatory changes including the low-gamma band during imitation. Twelve healthy, right-handed participants performed a finger task consisting of four conditions (imitation, execution, observation, and rest). During the imitation and execution conditions, significant event-related desynchronizations (ERDs) were observed at the left frontal, central, and parietal MEG sensors in the alpha, beta, and low-gamma bands. Functional connectivity analysis at the sensor level revealed an imitation-related connectivity between a group of frontal sensors and a group of parietal sensors in the low-gamma band. Furthermore, source reconstruction with synthetic aperture magnetometry showed significant ERDs in the low-gamma band in the left sensorimotor area and the middle frontal gyrus (MFG) during the imitation condition when compared with the other three conditions. Our results suggest that the oscillatory neural activities of the low-gamma band at the sensorimotor area and MFG play an important role in the observation-execution matching system related to imitation.

  3. Cyclin-dependent Kinase 8 Module Expression Profiling Reveals Requirement of Mediator Subunits 12 and 13 for Transcription of Serpent-dependent Innate Immunity Genes in Drosophila*

    PubMed Central

    Kuuluvainen, Emilia; Hakala, Heini; Havula, Essi; Sahal Estimé, Michelle; Rämet, Mika; Hietakangas, Ville; Mäkelä, Tomi P.

    2014-01-01

    The Cdk8 (cyclin-dependent kinase 8) module of Mediator integrates regulatory cues from transcription factors to RNA polymerase II. It consists of four subunits where Med12 and Med13 link Cdk8 and cyclin C (CycC) to core Mediator. Here we have investigated the contributions of the Cdk8 module subunits to transcriptional regulation using RNA interference in Drosophila cells. Genome-wide expression profiling demonstrated separation of Cdk8-CycC and Med12-Med13 profiles. However, transcriptional regulation by Cdk8-CycC was dependent on Med12-Med13. This observation also revealed that Cdk8-CycC and Med12-Med13 often have opposite transcriptional effects. Interestingly, Med12 and Med13 profiles overlapped significantly with that of the GATA factor Serpent. Accordingly, mutational analyses indicated that GATA sites are required for Med12-Med13 regulation of Serpent-dependent genes. Med12 and Med13 were also found to be required for Serpent-activated innate immunity genes in defense to bacterial infection. The results reveal a novel role for the Cdk8 module in Serpent-dependent transcription and innate immunity. PMID:24778181

  4. Kinase Activity Profiling of Pneumococcal Pneumonia

    PubMed Central

    Hoogendijk, Arie J.; Diks, Sander H.; van der Poll, Tom; Peppelenbosch, Maikel P.; Wieland, Catharina W.

    2011-01-01

    Background Pneumonia represents a major health burden. Previous work demonstrated that although the induction of inflammation is important for adequate host defense against pneumonia, an inability to regulate the host's inflammatory response within the lung later during infection can be detrimental. Intracellular signaling pathways commonly rely on activation of kinases, and kinases play an essential role in the regulation of the inflammatory response of immune cells. Methodology/Principal Findings Pneumonia was induced in mice via intranasal instillation of Streptococcus (S.) pneumoniae. Kinomics peptide arrays, exhibiting 1024 specific consensus sequences for protein kinases, were used to produce a systems biology analysis of cellular kinase activity during the course of pneumonia. Several differences in kinase activity revealed by the arrays were validated in lung homogenates of individual mice using western blot. We identified cascades of activated kinases showing that chemotoxic stress and a T helper 1 response were induced during the course of pneumococcal pneumonia. In addition, our data point to a reduction in WNT activity in lungs of S. pneumoniae infected mice. Moreover, this study demonstrated a reduction in overall CDK activity implying alterations in cell cycle biology. Conclusions/Significance This study utilizes systems biology to provide insight into the signaling events occurring during lung infection with the common cause of community acquired pneumonia, and may assist in identifying novel therapeutic targets in the treatment of bacterial pneumonia. PMID:21483672

  5. Generalized inverse analysis for fins of different profiles with all temperature-dependent parameters

    NASA Astrophysics Data System (ADS)

    Singh, Kuljeet; Das, Ranjan

    2016-08-01

    An inverse analysis is done to predict unknown and optimal dimensions of a fin satisfying either a given temperature or maximizing heat transfer rate. The profile simulating many geometries involves all temperature-dependent heat transfer modes. A hybrid algorithm is used to estimate relevant fin parameters. The present study shall be useful in selecting optimal dimensions to achieve either a particular temperature distribution or maximize heat transfer rate on various profiles.

  6. Activity-Dependent Model for Neuronal Avalanches

    NASA Astrophysics Data System (ADS)

    de Arcangelis, L.

    Networks of living neurons represent one of the most fascinating systems of modern biology. If the physical and chemical mechanisms at the basis of the functioning of a single neuron are quite well understood, the collective behavior of a system of many neurons is an extremely intriguing subject. Crucial ingredient of this complex behavior is the plasticity property of the network, namely the capacity to adapt and evolve depending on the level of activity. This plastic ability is believed, nowadays, to be at the basis of learning and memory in real brains. This fundamental problem in neurobiology has recently shown a number of features in common to other complex systems. These features mainly concern the morphology of the network, namely the spatial organization of the established connections, and a novel kind of neuronal activity. Experimental data have, in fact, shown that electrical information propagates in a cortex slice via an avalanche mode. Both features have been found in other problems in the context of the physics of complex systems and successful models have been developed to describe their behavior. In this contribution, we apply a statistical mechanical model to describe the complex activity in a neuronal network. The network is chosen to have a number of connections in long range, as found for neurons in vitro. The model implements the main physiological properties of living neurons and is able to reproduce recent experimental results. The numerical power spectra for electrical activity reproduces also the power law behavior measured in an EEG of man resting with the eyes closed.

  7. The multifaceted profile of activated microglia.

    PubMed

    Lynch, Marina A

    2009-10-01

    Although relatively neglected previously, research efforts in the past decade or so have identified a pivotal role for glial cells in regulating neuronal function. Particular emphasis has been placed on increasing our understanding of the function of microglia because a change from the ramified "resting" state of these cells has been associated with the pathogenesis of several neurodegenerative diseases, notably Alzheimer's disease. However, it is not clear whether activation of microglia and the associated inflammatory changes play a part in triggering disease processes or whether cell activation is a response to the early changes associated with the disease. In either case, the possibility exists that modulation of microglial activation may be beneficial in some circumstances, underlying the need to pursue research in this area. The original morphological categorization of microglia by Del Rio Hortega into ameboid, ramified, and intermediate forms, must now be elaborated to encompass a functional description. The evidence which has been generated recently suggests that microglia are probably never in a "resting" state and that several intermediate transitional states, based on function and morphology, probably exist. A more complete understanding of these states and the triggers which lead to a change from one to another state, and the factors which modulate the molecular switch that determines the persistence of the "activated" state remain to be identified.

  8. Comparison of Metalloproteinase Protein and Activity Profiling

    PubMed Central

    Giricz, Orsi; Lauer, Janelle L.; Fields, Gregg B.

    2010-01-01

    Proteolytic enzymes play fundamental roles in many biological processes. Members of the matrix metalloproteinase (MMP) family have been shown to take part in processes crucial in disease progression. The present study used the ExcelArray Human MMP/TIMP Array to quantify MMP and tissue inhibitor of metalloproteinase (TIMP) production in the lysates and media of 14 cancer and one normal cell line. The overall patterns were very similar in terms of which MMPs and TIMPs were secreted in the media versus associated with the cells in the individual samples. However, more MMP was found in the media, both in amount and in variety. TIMP-1 was produced in all cell lines. MMP activity assays with three different FRET substrates were then utilized to determine if protein production correlated with function for the WM-266-4 and BJ cell lines. Metalloproteinase activity was observed for both cell lines with a general MMP substrate (Knight SSP), consistent with protein production data. However, although both cell lines promoted the hydrolysis of a more selective MMP substrate (NFF-3), metalloproteinase activity was only confirmed in the BJ cell line. The use of inhibitors to confirm metalloproteinase activities pointed to the strengths and weaknesses of in situ FRET substrate assays. PMID:20920458

  9. Criminal profiling as a plotting activity based on abductive processes.

    PubMed

    Verde, Alfredo; Nurra, Antonio

    2010-10-01

    In this article the authors analyze the nature and aims of criminal profiling from a theoretical point of view. The need to become increasingly "scientific" has given rise to the modern approaches of profiling, which have been particularly successful in cases of serial homicides and sex crimes, given that compulsive (perverse) acts, because of their ritual nature, have been described as being more easily foreseeable and presumably linkable to the psychological and even personal characteristics of a given criminal. On this basis, the authors analyze profiling from an epistemological point of view and show how, in the concrete activity of profiling, profilers depart from the "certainty" of the scientific models (those that are based on deductive-inductive processes); the epistemological basis of reasoning changes as there is no longer an induction-deduction model but rather an abductive model (as conceived and explained by Peirce) in which the importance of plotting (the weaving of a narrative) becomes greater.

  10. Neighborhood environment profiles related to physical activity and weight status: a latent profile analysis.

    PubMed

    Adams, Marc A; Sallis, James F; Kerr, Jacqueline; Conway, Terry L; Saelens, Brian E; Frank, Lawrence D; Norman, Gregory J; Cain, Kelli L

    2011-05-01

    Neighborhood built environments (BE) include combinations of co-existing stimuli influencing physical activity (PA). Dealing with numerous environmental variables and complexity presents a significant challenge. The current analysis explored whether a range of reported BE features associated with adults' physical activity produced distinct multivariate patterns, and tested whether adults' PA and body mass differed by BE profiles. Participants (20-65 years, 48.2% female, 26% ethnic minority) were recruited between 2002 and 2005 from 32 neighborhoods from Seattle-King County, WA (N=1287) and Baltimore, MD-Washington, DC regions (N=912). Independent Latent Profile Analyses were conducted in each region with 11 environmental variables from the Neighborhood Environment Walkability Scale. Validity of the neighborhood profiles was examined by their relationship to PA (accelerometer-derived moderate-to-vigorous minutes/day, self-reported minutes/week of walking for transportation and leisure) and self-reported BMI using ANCOVA models. Neighborhood profiles for Seattle and Baltimore regions were visually similar, suggesting generalizability. High-walkable recreationally-dense neighborhoods differed significantly from other neighborhood types by as much as 13 MVPA minutes/day, almost 60 minutes/week of walking for transportation, and 75 min/week of leisure-time activity. Neighborhood profiles also differed significantly for BMI. These findings could help identify optimal patterns of environmental attributes that facilitate physical activity and improve weight status. Copyright © 2010 Elsevier Inc. All rights reserved.

  11. A Profile of Latino School-Based Extracurricular Activity Involvement

    ERIC Educational Resources Information Center

    Peguero, Anthony A.

    2010-01-01

    Participation in school-based extracurricular activities influences educational success. Thus, it is important to depict a profile of school-based extracurricular activity involvement for a Latino student population that is marginalized in schools. This research uses the Educational Longitudinal Study of 2002 and logistic regression analyses to…

  12. Lunar seismic profiling experiment natural activity study

    NASA Technical Reports Server (NTRS)

    Duennebier, F. K.

    1976-01-01

    The Lunar Seismic Experiment Natural Activity Study has provided a unique opportunity to study the high frequency (4-20 Hz) portion to the seismic spectrum on the moon. The data obtained from the LSPE was studied to evaluate the origin and importance of the process that generates thermal moonquakes and the characteristics of the seismic scattering zone at the lunar surface. The detection of thermal moonquakes by the LSPE array made it possible to locate the sources of many events and determine that they are definitely not generated by astronaut activities but are the result of a natural process on the moon. The propagation of seismic waves in the near-surface layers was studied in a qualitative manner. In the absence of an adequate theoretical model for the propagation of seismic waves in the moon, it is not possible to assign a depth for the scattering layer. The LSPE data does define several parameters which must be satisfied by any model developed in the future.

  13. Variability of humic acid properties depending on their precursor material: a study of peat profiles

    NASA Astrophysics Data System (ADS)

    Klavins, Maris; Purmalis, Oskars

    2015-04-01

    Analysis of variation of peat composition, presence of trace elements in the peat and HSs within peat profiles can give information on the character of transformation of organic matter, important for C biogeochemical cycling, but also about impacts of climate change and human activities. In peat the transformation and decay process of living organic matter (humification) is retarded by the acidic and anaerobic environment, but at the same time the peat can provide information on environmental and paleo-environmental conditions of the past. The aim of the present study is to analyze the elemental and functional composition, spectral characteristics of humic acids isolated from a well characterized raised bog peat profiles to evaluate the impact of the character of humification processes on the peat HA properties. A comparative and complex characterization of humic acids (HAs) isolated from peat profiles of different origin in Latvia was carried out. Elemental and functional analysis of the isolated HAs was done, their acidity and molecular weights estimated. Spectral characterization included UV-Vis, IR, and electron spin resonance and fluorescence spectra. Structural characterization of HAs was by both 1H and 13C nuclear magnetic resonance spectra. Comparison of position of studied humic acids in the Van Krevelen graph was done, thus locating them in the biogeochemical transformation processes of organic matter. Properties of HAs isolated from the Latvian peat were compared with HA from other sources (soil, water, coal and synthetic humic substances). Major properties of peat HAs depended on their origin, indicating the importance of humification processes. HAs isolated from peat of more recent origin were more similar to soil HAs, but from older sources there was a greater degree of humification. Changes of surface tension of solutions of humic acids stress the differences in aggregation character - ability to form supramacromolecular complexes of humic substances

  14. Comprehensive Analysis of PPARα-Dependent Regulation of Hepatic Lipid Metabolism by Expression Profiling

    PubMed Central

    Rakhshandehroo, Maryam; Sanderson, Linda M.; Matilainen, Merja; Stienstra, Rinke; Carlberg, Carsten; de Groot, Philip J.; Müller, Michael; Kersten, Sander

    2007-01-01

    PPARα is a ligand-activated transcription factor involved in the regulation of nutrient metabolism and inflammation. Although much is already known about the function of PPARα in hepatic lipid metabolism, many PPARα-dependent pathways and genes have yet to be discovered. In order to obtain an overview of PPARα-regulated genes relevant to lipid metabolism, and to probe for novel candidate PPARα target genes, livers from several animal studies in which PPARα was activated and/or disabled were analyzed by Affymetrix GeneChips. Numerous novel PPARα-regulated genes relevant to lipid metabolism were identified. Out of this set of genes, eight genes were singled out for study of PPARα-dependent regulation in mouse liver and in mouse, rat, and human primary hepatocytes, including thioredoxin interacting protein (Txnip), electron-transferring-flavoprotein β polypeptide (Etfb), electron-transferring-flavoprotein dehydrogenase (Etfdh), phosphatidylcholine transfer protein (Pctp), endothelial lipase (EL, Lipg), adipose triglyceride lipase (Pnpla2), hormone-sensitive lipase (HSL, Lipe), and monoglyceride lipase (Mgll). Using an in silico screening approach, one or more PPAR response elements (PPREs) were identified in each of these genes. Regulation of Pnpla2, Lipe, and Mgll, which are involved in triglyceride hydrolysis, was studied under conditions of elevated hepatic lipids. In wild-type mice fed a high fat diet, the decrease in hepatic lipids following treatment with the PPARα agonist Wy14643 was paralleled by significant up-regulation of Pnpla2, Lipe, and Mgll, suggesting that induction of triglyceride hydrolysis may contribute to the anti-steatotic role of PPARα. Our study illustrates the power of transcriptional profiling to uncover novel PPARα-regulated genes and pathways in liver. PMID:18288265

  15. Comprehensive analysis of PPARalpha-dependent regulation of hepatic lipid metabolism by expression profiling.

    PubMed

    Rakhshandehroo, Maryam; Sanderson, Linda M; Matilainen, Merja; Stienstra, Rinke; Carlberg, Carsten; de Groot, Philip J; Müller, Michael; Kersten, Sander

    2007-01-01

    PPARalpha is a ligand-activated transcription factor involved in the regulation of nutrient metabolism and inflammation. Although much is already known about the function of PPARalpha in hepatic lipid metabolism, many PPARalpha-dependent pathways and genes have yet to be discovered. In order to obtain an overview of PPARalpha-regulated genes relevant to lipid metabolism, and to probe for novel candidate PPARalpha target genes, livers from several animal studies in which PPARalpha was activated and/or disabled were analyzed by Affymetrix GeneChips. Numerous novel PPARalpha-regulated genes relevant to lipid metabolism were identified. Out of this set of genes, eight genes were singled out for study of PPARalpha-dependent regulation in mouse liver and in mouse, rat, and human primary hepatocytes, including thioredoxin interacting protein (Txnip), electron-transferring-flavoprotein beta polypeptide (Etfb), electron-transferring-flavoprotein dehydrogenase (Etfdh), phosphatidylcholine transfer protein (Pctp), endothelial lipase (EL, Lipg), adipose triglyceride lipase (Pnpla2), hormone-sensitive lipase (HSL, Lipe), and monoglyceride lipase (Mgll). Using an in silico screening approach, one or more PPAR response elements (PPREs) were identified in each of these genes. Regulation of Pnpla2, Lipe, and Mgll, which are involved in triglyceride hydrolysis, was studied under conditions of elevated hepatic lipids. In wild-type mice fed a high fat diet, the decrease in hepatic lipids following treatment with the PPARalpha agonist Wy14643 was paralleled by significant up-regulation of Pnpla2, Lipe, and Mgll, suggesting that induction of triglyceride hydrolysis may contribute to the anti-steatotic role of PPARalpha. Our study illustrates the power of transcriptional profiling to uncover novel PPARalpha-regulated genes and pathways in liver.

  16. Cholesterol-dependent hemolytic activity of Passiflora quadrangularis leaves.

    PubMed

    Yuldasheva, L N; Carvalho, E B; Catanho, M-T J A; Krasilnikov, O V

    2005-07-01

    Plants used in traditional medicine are rich sources of hemolysins and cytolysins, which are potential bactericidal and anticancer drugs. The present study demonstrates for the first time the presence of a hemolysin in the leaves of Passiflora quadrangularis L. This hemolysin is heat stable, resistant to trypsin treatment, has the capacity to froth, and acts very rapidly. The hemolysin activity is dose-dependent, with a slope greater than 1 in a double-logarithmic plot. Polyethylene glycols of high molecular weight were able to reduce the rate of hemolysis, while liposomes containing cholesterol completely inhibited it. In contrast, liposomes containing phosphatidylcholine were ineffective. The Passiflora hemolysin markedly increased the conductance of planar lipid bilayers containing cholesterol but was ineffective in cholesterol-free bilayers. Successive extraction of the crude hemolysin with n-hexane, chloroform, ethyl acetate, and n-butanol resulted in a 10-fold purification, with the hemolytic activity being recovered in the n-butanol fraction. The data suggest that membrane cholesterol is the primary target for this hemolysin and that several hemolysin molecules form a large transmembrane water pore. The properties of the Passiflora hemolysin, such as its frothing ability, positive color reaction with vanillin, selective extraction with n-butanol, HPLC profile, cholesterol-dependent membrane susceptibility, formation of a stable complex with cholesterol, and rapid erythrocyte lysis kinetics indicate that it is probably a saponin.

  17. P/Halley - Effects of time-dependent production rates on spatial emission profiles

    NASA Astrophysics Data System (ADS)

    Combi, M. R.; Fink, U.

    1993-06-01

    Spatial profiles of C2, CN, NH2, and O(D-1) in Comet P/Halley taken on 1986 March 1.54 and 2.55 and April 14.32 and 15.30 clearly show the effect of the 7 day periodic variation seen in photometric observations. With a time-dependent model based upon the light curve and employing standard scale lengths for each species reduced to the appropriate heliocentric distance, we are able to reproduce the highly variable profiles for all species. We computed the phase lag and amplitude correction between the actual gas production at the nucleus and the temporal/spatial filter imposed by the finite aperture photometry. For early March we find a phase lag and amplitude correction of 12 hr and -9 percent, respectively, whereas in mid-April the values are 6 hr and -27 percent. The same phase lag and amplitude correction work equally well for all four species despite their wide variation in photochemical lifetimes for production and decay. The same model integrated over circular apertures is able to reproduce the entire published March and April photometric light curves for C2. Our results require the use of the 7.60 day period for the March data as opposed to the 7.37 day period that is relevant for the April data, in agreement with the published analysis of the photometric data. Our results will help to reconcile the placement of active areas on the surface of Halley's comet with various remote observations and spacecraft images of the nucleus.

  18. Quantitative genetic activity graphical profiles for use in chemical evaluation

    SciTech Connect

    Waters, M.D.; Stack, H.F.; Garrett, N.E.; Jackson, M.A.

    1990-12-31

    A graphic approach, terms a Genetic Activity Profile (GAP), was developed to display a matrix of data on the genetic and related effects of selected chemical agents. The profiles provide a visual overview of the quantitative (doses) and qualitative (test results) data for each chemical. Either the lowest effective dose or highest ineffective dose is recorded for each agent and bioassay. Up to 200 different test systems are represented across the GAP. Bioassay systems are organized according to the phylogeny of the test organisms and the end points of genetic activity. The methodology for producing and evaluating genetic activity profile was developed in collaboration with the International Agency for Research on Cancer (IARC). Data on individual chemicals were compiles by IARC and by the US Environmental Protection Agency (EPA). Data are available on 343 compounds selected from volumes 1-53 of the IARC Monographs and on 115 compounds identified as Superfund Priority Substances. Software to display the GAPs on an IBM-compatible personal computer is available from the authors. Structurally similar compounds frequently display qualitatively and quantitatively similar profiles of genetic activity. Through examination of the patterns of GAPs of pairs and groups of chemicals, it is possible to make more informed decisions regarding the selection of test batteries to be used in evaluation of chemical analogs. GAPs provided useful data for development of weight-of-evidence hazard ranking schemes. Also, some knowledge of the potential genetic activity of complex environmental mixtures may be gained from an assessment of the genetic activity profiles of component chemicals. The fundamental techniques and computer programs devised for the GAP database may be used to develop similar databases in other disciplines. 36 refs., 2 figs.

  19. Statistical assessment of dissolution and drug release profile similarity using a model-dependent approach.

    PubMed

    Berry, Mark R; Likar, Michael D

    2007-10-18

    A general multivariate procedure for assessing the similarity of dissolution and drug release profiles was developed. A mathematical model is fit to the data, and Hotelling's T(2) test is used to calculate the joint confidence region around the vector of differences between least-squares estimates of the parameters in the model. The method of Lagrange multipliers is used to determine if this confidence region is enclosed within a predetermined similarity region, and profile similarity is claimed if this is the case. The first-order, Gompertz, logistic, second-order, and Weibull models were fit to the in vitro extended-release profile of pseudoephedrine HCl from an asymmetric membrane (AM) film-coated osmotic tablet. The first-order model was selected because of its simplicity and because it was the best-fitting model according to a modified form of Akaike's Information Criterion. A nonlinear response surface model was also developed so that the formulator could calculate how much of the AM film coat should be applied in order to obtain the desired drug release profile. The usefulness of this model-dependent procedure was further demonstrated during an analytical method transfer exercise, where it was used to compare the drug release profiles obtained by two independent laboratories; additional research is required, however, before the appropriate acceptance criteria for demonstrating profile similarity can be recommended.

  20. Influence of carrier flow on the temperature-dependent capacitance-voltage profiles of heterojunction structures

    NASA Astrophysics Data System (ADS)

    Kwon, S. D.; Lim, H.; Shin, H. K.; Choe, Byung-Doo

    1996-10-01

    We suggest a model which can explain the shifting of carrier concentration peaks in the temperature-dependent capacitance-voltage carrier profiles of heterojunction (HJ) structures. The shift of concentration peaks, which was frequently observed in the inverted isotype HJs was previously attributed to the traps at the heterointerface. The main feature of our model is the role of band offset as a limiter to the test signal current. The model can explain the difference of the peak shift in the carrier profiles of the normal and inverted type HJs. According to this model, the peak shifts at low temperatures occur naturally for the inverted type HJs.

  1. Distinct transcriptome profiles differentiate NSAID-dependent from NSAID-independent food anaphylaxis

    PubMed Central

    Muñoz-Cano, Rosa; Pascal, Mariona; Bartra, Joan; Picado, Cesar; Valero, Antonio; Kim, Do-Kyun; Brooks, Stephen; Ombrello, Michael; Metcalfe, Dean D.; Rivera, Juan; Olivera, Ana

    2015-01-01

    Background Lipid transfer protein (LTP), an abundant protein in fruits, vegetables and nuts, is a common food allergen in Mediterranean areas causing diverse allergic reactions. Approximately 40% of food anaphylaxis induced by LTP require non-steroidal anti-inflammatory drugs (NSAIDs) as a triggering cofactor. Objective To better understand the determinants of NSAID-dependent (NSAID-LTP-A) and NSAID-independent LTP-anaphylaxis (LTP-A) Methods Selection of patients was based on a proven clinical history of NSAID-dependent or -independent anaphylaxis to LTP, positive skin prick test to LTP and serum LTP-IgE. Whole transcriptome (RNA-Seq) analysis of blood cells from 14 individuals with NSAID-LTP-A, 7 with LTP-A and 13 healthy controls was performed to identify distinct gene expression signatures. Results Expression of genes regulating gastrointestinal epithelium renewal was altered in both patient sets, particularly in LTP-A, who also presented gene expression profiles characteristic of an inflammatory syndrome. These included altered B cell pathways, increased neutrophil activation markers and elevated levels of reactive oxygen species. Increased expression of the IgG receptor (CD64) in LTP-A patients was mirrored by the presence of LTP-specific IgG1 and 3. Conversely, NSAID-LTP-A patients were characterized by reduced expression of IFN-γ-regulated genes and IFN-γ levels as well as up-regulated adenosine receptor 3 (ADORA3) expression and genes related to adenosine metabolism. Conclusions Gene ontology analysis suggests disturbances in gut epithelium homeostasis in both LTP-related anaphylaxis groups with potential integrity breaches in LTP-A that may explain their distinct inflammatory signature. Differential regulation in LTP-A and NSAID-LTP-A of the IFN-γ pathway, IgG receptors and ADORA3 may provide the pathogenic basis of their distinct responses. PMID:26194548

  2. Activity-based profiling of the proteasome pathway during hepatitis C virus infection.

    PubMed

    Nasheri, Neda; Ning, Zhibin; Figeys, Daniel; Yao, Shao; Goto, Natalie K; Pezacki, John Paul

    2015-11-01

    Hepatitis C virus (HCV) infection often leads to chronic hepatitis, liver cirrhosis, and hepatocellular carcinoma. The stability of the HCV proteins is controlled by ubiquitin-dependent and ubiquitin-independent proteasome pathways. Many viruses modulate proteasome function for their propagation. To examine the interrelationship between HCV and the proteasome pathways we employed a quantitative activity-based protein profiling method. Using this approach we were able to quantify the changes in the activity of several proteasome subunits and found that proteasome activity is drastically reduced by HCV replication. The results imply a link between the direct downregulation of the activity of this pathway and chronic HCV infection.

  3. Distinct Metal Isoforms Underlie Promiscuous Activity Profiles of Metalloenzymes.

    PubMed

    Baier, Florian; Chen, John; Solomonson, Matthew; Strynadka, Natalie C J; Tokuriki, Nobuhiko

    2015-07-17

    Within a superfamily, functionally diverged metalloenzymes often favor different metals as cofactors for catalysis. One hypothesis is that incorporation of alternative metals expands the catalytic repertoire of metalloenzymes and provides evolutionary springboards toward new catalytic functions. However, there is little experimental evidence that incorporation of alternative metals changes the activity profile of metalloenzymes. Here, we systematically investigate how metals alter the activity profiles of five functionally diverged enzymes of the metallo-β-lactamase (MBL) superfamily. Each enzyme was reconstituted in vitro with six different metals, Cd(2+), Co(2+), Fe(2+), Mn(2+), Ni(2+), and Zn(2+), and assayed against eight catalytically distinct hydrolytic reactions (representing native functions of MBL enzymes). We reveal that each enzyme metal isoform has a significantly different activity level for native and promiscuous reactions. Moreover, metal preferences for native versus promiscuous activities are not correlated and, in some cases, are mutually exclusive; only particular metal isoforms disclose cryptic promiscuous activities but often at the expense of the native activity. For example, the L1 B3 β-lactamase displays a 1000-fold catalytic preference for Zn(2+) over Ni(2+) for its native activity but exhibits promiscuous thioester, phosphodiester, phosphotriester, and lactonase activity only with Ni(2+). Furthermore, we find that the five MBL enzymes exist as an ensemble of various metal isoforms in vivo, and this heterogeneity results in an expanded activity profile compared to a single metal isoform. Our study suggests that promiscuous activities of metalloenzymes can stem from an ensemble of metal isoforms in the cell, which could facilitate the functional divergence of metalloenzymes.

  4. [Peculiarities of cardiovascular system pathology depending on psychological profile in patients of senior age groups].

    PubMed

    Prokhorenko, I O

    2013-01-01

    Interrelations between peculiarities of psychological profile of patients of senior age groups (according to Cattel), level of stress hormones in blood and background pathology of cardiovascular system were studied. Levels of catecholamine and corticosteroids in dynamics, rate of magnesium in erythrocytes and calcium in plaques of coronary arteries as well as fats, Holter ECG, daily profiles of blood pressure, vasomotor function of endothelium and microcirculation were analysed. It is established that stress hormones indirectly determine original form of stress reaction depending on patients' psychological profile. This contributes to the development of one or another form of cardiovascular system pathology. Excessive alcohol intake also promotes progression of cardiovascular system pathology. Depression, being a reflection of disbalance of stress hormones levels, can be used as a marker of unfavourable course of cardiovascular pathology.

  5. Antiorthostatic suspension stimulates profiles of macrophage activation in mice

    NASA Technical Reports Server (NTRS)

    Miller, E. S.; Bates, R. A.; Koebel, D. A.; Sonnenfeld, G.

    1999-01-01

    The antiorthostatic suspension model simulates certain physiological effects of spaceflight. We have previously reported BDF1 mice suspended by the tail in the antiorthostatic orientation for 4 days express high levels of resistance to virulent Listeria monocytogenesinfection. In the present study, we examined whether the increased resistance to this organism correlates with profiles of macrophage activation, given the role of the macrophage in killing this pathogen in vivo. We infected BDF1 mice with a lethal dose of virulent L. monocytogenes on day 4 of antiorthostatic suspension and 24 h later constructed profiles of macrophage activation. Viable listeria could not be detected in mice suspended in the antiorthostatic orientation 24 h after infection. Flow cytometric analysis revealed the numbers of granulocytes and mononuclear phagocytes in the spleen of infected mice were not significantly altered as a result of antiorthostatic suspension. Splenocytes from antiorthostatically suspended infected mice produced increased titers of IL-1. Serum levels of neopterin, a nucleotide metabolite secreted by activated macrophages, were enhanced in mice infected during antiorthostatic suspension, but not in antiorthostatically suspended naive mice. Splenic macrophages from mice infected on day 4 of suspension produced enhanced levels of lysozyme. In contrast to the results from antiorthostatically suspended infected mice, macrophages from antiorthostatically suspended uninfected mice did not express enhanced bactericidal activities. The collective results indicate that antiorthostatic suspension can stimulate profiles of macrophage activation which correlate with increased resistance to infection by certain classes of pathogenic bacteria.

  6. Antiorthostatic suspension stimulates profiles of macrophage activation in mice

    NASA Technical Reports Server (NTRS)

    Miller, E. S.; Bates, R. A.; Koebel, D. A.; Sonnenfeld, G.

    1999-01-01

    The antiorthostatic suspension model simulates certain physiological effects of spaceflight. We have previously reported BDF1 mice suspended by the tail in the antiorthostatic orientation for 4 days express high levels of resistance to virulent Listeria monocytogenesinfection. In the present study, we examined whether the increased resistance to this organism correlates with profiles of macrophage activation, given the role of the macrophage in killing this pathogen in vivo. We infected BDF1 mice with a lethal dose of virulent L. monocytogenes on day 4 of antiorthostatic suspension and 24 h later constructed profiles of macrophage activation. Viable listeria could not be detected in mice suspended in the antiorthostatic orientation 24 h after infection. Flow cytometric analysis revealed the numbers of granulocytes and mononuclear phagocytes in the spleen of infected mice were not significantly altered as a result of antiorthostatic suspension. Splenocytes from antiorthostatically suspended infected mice produced increased titers of IL-1. Serum levels of neopterin, a nucleotide metabolite secreted by activated macrophages, were enhanced in mice infected during antiorthostatic suspension, but not in antiorthostatically suspended naive mice. Splenic macrophages from mice infected on day 4 of suspension produced enhanced levels of lysozyme. In contrast to the results from antiorthostatically suspended infected mice, macrophages from antiorthostatically suspended uninfected mice did not express enhanced bactericidal activities. The collective results indicate that antiorthostatic suspension can stimulate profiles of macrophage activation which correlate with increased resistance to infection by certain classes of pathogenic bacteria.

  7. Chemoproteomic profiling of host and pathogen enzymes active in cholera

    PubMed Central

    Hatzios, Stavroula K.; Hubbard, Troy; Sasabe, Jumpei; Munera, Diana; Clark, Lars; Bachovchin, Daniel A.; Qadri, Firdausi; Ryan, Edward T.; Davis, Brigid M.; Weerapana, Eranthie; Waldor, Matthew K.

    2016-01-01

    Activity-based protein profiling (ABPP) is a chemoproteomic tool for detecting active enzymes in complex biological systems. We used ABPP to identify secreted bacterial and host serine hydrolases that are active in animals infected with the cholera pathogen Vibrio cholerae. Four V. cholerae proteases were consistently active in infected rabbits, and one, VC0157 (renamed IvaP), was also active in human cholera stool. Inactivation of IvaP influenced the activity of other secreted V. cholerae and rabbit enzymes in vivo, while genetic disruption of all four proteases increased the abundance and binding of an intestinal lectin—intelectin—to V. cholerae in infected rabbits. Intelectin also bound to other enteric bacterial pathogens, suggesting it may constitute a previously unrecognized mechanism of bacterial surveillance in the intestine that is inhibited by pathogen-secreted proteases. Our work demonstrates the power of activity-based proteomics to reveal host-pathogen enzymatic dialogue in an animal model of infection. PMID:26900865

  8. Chemoproteomic profiling of host and pathogen enzymes active in cholera.

    PubMed

    Hatzios, Stavroula K; Abel, Sören; Martell, Julianne; Hubbard, Troy; Sasabe, Jumpei; Munera, Diana; Clark, Lars; Bachovchin, Daniel A; Qadri, Firdausi; Ryan, Edward T; Davis, Brigid M; Weerapana, Eranthie; Waldor, Matthew K

    2016-04-01

    Activity-based protein profiling (ABPP) is a chemoproteomic tool for detecting active enzymes in complex biological systems. We used ABPP to identify secreted bacterial and host serine hydrolases that are active in animals infected with the cholera pathogen Vibrio cholerae. Four V. cholerae proteases were consistently active in infected rabbits, and one, VC0157 (renamed IvaP), was also active in human choleric stool. Inactivation of IvaP influenced the activity of other secreted V. cholerae and rabbit enzymes in vivo, and genetic disruption of all four proteases increased the abundance of intelectin, an intestinal lectin, and its binding to V. cholerae in infected rabbits. Intelectin also bound to other enteric bacterial pathogens, suggesting that it may constitute a previously unrecognized mechanism of bacterial surveillance in the intestine that is inhibited by pathogen-secreted proteases. Our work demonstrates the power of activity-based proteomics to reveal host-pathogen enzymatic dialog in an animal model of infection.

  9. Human corneal epithelial subpopulations: oxygen dependent ex vivo expansion and transcriptional profiling.

    PubMed

    Bath, Chris

    2013-06-01

    Corneal epithelium is being regenerated throughout life by limbal epithelial stem cells (LESCs) believed to be located in histologically defined stem cell niches in corneal limbus. Defective or dysfunctional LESCs result in limbal stem cell deficiency (LSCD) causing pain and decreased visual acuity. Since the first successful treatment of LSCD by transplantation of ex vivo expanded LESCs in 1997, many attempts have been carried out to optimize culture conditions to improve the outcome of surgery. To date, progress in this field of bioengineering is substantially hindered by both the lack of specific biomarkers of LESCs and the lack of a precise molecular characterization of in situ epithelial subpopulations. The aim of this dissertation was to optimize culture systems with regard to the environmental oxygen concentration for selective ex vivo expansion of LESCs and to analyse in situ subpopulations in human corneal epithelium using a combination of laser capture microdissection and RNA sequencing for global transcriptomic profiling. We compared dissociation cultures, using either expansion on γ-irradiated NIH/3T3 feeder cells in serum-rich medium or expansion directly on plastic in serum-free EpiLife medium, using a range of physiologically relevant oxygen concentrations (2%, 5%, 10%, 15% and 20%). Using immunocytochemistry and advanced fluorescence microscopy, cells were characterized regarding growth, cell cycle distribution, colony-forming efficiency (CFE), phenotypes and cytomorphometry. Limbal epithelial cells expanded in 2% O2 exhibited slow growth, low fraction of cells in S/G2 , high CFE, high expression of stem cell markers ABCG2 and p63α, and low fraction of differentiation marker CK3 resembling a LESC phenotype. The effect of hypoxia to maintain LESCs in culture was not dependent on the system used for propagation (Bath et al. 2013a). Laser capture microdissection was used to isolate cellular subpopulations in situ from the spatially defined

  10. Propagation-dependent beam profile distortion associated with the Goos-Hanchen shift.

    PubMed

    Wan, Yuhang; Zheng, Zheng; Zhu, Jinsong

    2009-11-09

    The propagation-dependent profile distortion of the reflected beam is studied via deriving the theoretical model of the optical field distribution in both the near and far field. It is shown that strong and fast-varying beam distortions can occur along the propagation path, compared to the profile on the reflecting surface. Numerical simulations for the case of a typical SPR configuration with a sharp angular response curve reveal that, when the phase distribution in the angular range covered by the input beam becomes nonlinear, previous theories based on the linear phase approximation fail to predict the Goos-Hanchen shift and its propagation-dependent variations precisely. Our study could shed light on more accurate modeling of the Goos-Hanchen effect's impact on the relevant photonic devices and measurement applications.

  11. Time-dependent analysis of 8 days of CN spatial profiles in comet P/Halley

    NASA Technical Reports Server (NTRS)

    Combi, Michael; Huang, Bormin; Cochran, Anita; Fink, Uwe; Schulz, Rita

    1994-01-01

    CN profiles in comet P/Halley were constructed from observations taken at three observatories during an 8 day period in April 1986. These data provide a time series of CN spatial profiles spanning just over one 7.37 day period from 1986 April 7 to April 15 and sample distances from the nucleus from just over 10(exp 3) km to 10(exp 6) km. The effect of the 7.37 day periodic variation on the CN distribution in P/Halley has been examined by using the time-dependent model applied earlier to a subset of the data. Because of the large spatial scale of the data on April 7, 8, and 9 (approx. 10(exp 6) km), and the corresponding transport time in the coma, information present in the spatial profiles regarding the gas production rate actually covers nearly two full periods. These spatially extended profiles clearly show the wavy structures outside 10(exp 5) km. Such structures were predicted in a previous analysis (Combi & Fink 1993) that was based solely on the photometric light curve and on profiles which only extended to distances less than 10(exp 5) km. We are now able to reproduce the highly variable Halley correction for the variation in gas production rate.

  12. Power-law velocity profile in turbulent boundary layers: An integral reynolds-number dependent solution

    NASA Astrophysics Data System (ADS)

    Castro-Orgaz, Oscar; Dey, Subhasish

    2011-10-01

    Geophysical flows of practical interest encompass turbulent boundary layer flows. The velocity profile in turbulent flows is generally described by a log- or a power-law applicable to certain zones of the boundary layer, or by wall-wake law for the entire zone of the boundary layer. In this study, a novel theory is proposed from which the power-law velocity profile is obtained for the turbulent boundary layer flow. The new power-law profile is based on the conservation of mass and the skin friction within the boundary layer. From the proposed theory, analytical expressions for the power-law velocity profile are presented, and their Reynolds-number dependency is highlighted. The velocity profile, skin friction coefficient and boundary layer thickness obtained from the proposed theory are validated by the reliable experimental data for zero-pressure gradient turbulent boundary layers. The expressions for Reynolds shear stress and eddy viscosity distributions across the boundary layer are also obtained and validated by the experimental data.

  13. A new instrument to measure the activity profile of elderly shoulder pathology patients: the Senior Shoulder Activity score (SSA score).

    PubMed

    Deranlot, J; Flurin, P-H; Hardy, P; Klouche, S

    2013-12-01

    Whether rotator cuff repair is indicated in an elderly subject depends on the patient's activity profile and functional demand. A Senior Shoulder Activity (SSA) score is described, as a support for indications and analysis of clinical results according to activity level. The SSA score, comprising 4 levels from "sedentary" to "very active", was validated by comparison against a control group of 113 asymptomatic patients. It was included in the protocol of the French Arthroscopy Society's comparative study of repair versus simple decompression in 143 rotator cuff tears. Recovery of activity was assessed according to procedure. At 1-year follow-up, suturing was associated with recovery of previous activity level in 87% of the cases and in 80% for decompression, a non-significant difference. When, however, less active patients (SSA 1 and 2) were contrasted with the more active (SSA 3 and 4), clinical results with suture versus decompression on Constant score showed a greater difference in the SSA 3-4 group. The SSA score is not the same as the activity item of the Constant score, as it assesses the patient's usual activity level, before symptom onset, whereas the Constant item assesses activity at a given moment, independently of the patient's normal activity profile. The Senior Shoulder Activity score is a simple, reproducible complement to the Constant score, revealing differences in clinical results on the latter, according to activity profile. Rotator cuff repair or simple decompression provided recovery of previous SSA activity level in more than 80% of the cases. The difference in clinical results between the two was significantly greater in more active patients. It would seem to follow that suture is more beneficial for more active subjects while simple decompression may be suitable for those with lower functional demand. Copyright © 2013. Published by Elsevier Masson SAS.

  14. Activity profile of high-level Australian lacrosse players.

    PubMed

    Polley, Chris S; Cormack, Stuart J; Gabbett, Tim J; Polglaze, Ted

    2015-01-01

    Despite lacrosse being one of the fastest growing team sports in the world, there is a paucity of information detailing the activity profile of high-level players. Microtechnology systems (global positioning systems and accelerometers) provide the opportunity to obtain detailed information on the activity profile in lacrosse. Therefore, this study aimed to analyze the activity profile of lacrosse match-play using microtechnology. Activity profile variables assessed relative to minutes of playing time included relative distance (meter per minute), distance spent standing (0-0.1 m·min), walking (0.2-1.7 m·min), jogging (1.8-3.2 m·min), running (3.3-5.6 m·min), sprinting (≥5.7 m·min), number of high, moderate, low accelerations and decelerations, and player load (PL per minute), calculated as the square root of the sum of the squared instantaneous rate of change in acceleration in 3 vectors (medio-lateral, anterior-posterior, and vertical). Activity was recorded from 14 lacrosse players over 4 matches during a national tournament. Players were separated into positions of attack, midfield, or defense. Differences (effect size [ES] ± 90% confidence interval) between positions and periods of play were considered likely positive when there was ≥75% likelihood of the difference exceeding an ES threshold of 0.2. Midfielders had likely covered higher (mean ± SD) meters per minute (100 ± 11) compared with attackers (87 ± 14; ES = 0.89 ± 1.04) and defenders (79 ± 14; ES = 1.54 ± 0.94) and more moderate and high accelerations and decelerations. Almost all variables across positions were reduced in quarter 4 compared with quarter 1. Coaches should accommodate for positional differences when preparing lacrosse players for competition.

  15. Growth phase-dependent variations in transcript profiles for thioredoxin- and glutathione-dependent redox systems followed by budding and hyphal Candida albicans cultures.

    PubMed

    Michán, Carmen; Pueyo, Carmen

    2009-10-01

    We report the absolute transcription profiles of 24 genes coding for putative thioredoxin (Trx)- and glutathione (GSH)-dependent redox system components, accompanying the Candida albicans batch culture growth, under either yeast or hyphal conditions. All mRNAs investigated (plus the housekeeping ACT1) displayed significant alterations in their steady-state copy number. Collectively, these quantifications show that: (1) the most abundant mRNAs during active growth coded for putative thiol peroxidases (TSA1, PRX1, AHP11 and AHP12) and for donor thioredoxin Trx1p, i.e. those five mRNAs represented >74% of all transcript molecules quantified in a late exponential phase; (2) the transcripts under study differed between budding and hyphal cells not only in their abundance but also in their profiles throughout the growth stages; (3) mRNA amounts for four GSH-transferase putative genes (GTT12, GTT13, GTT14 and GST3) increased in the stationary phase in yeast but not under filamentous conditions. Hydrogen peroxide resistance, plus GSH, GSSG and reactive oxygen species contents, throughout yeast and hyphal growth, were also studied, and the differences observed were related to the transcript profiles.

  16. Active force perception depends on cerebellar function.

    PubMed

    Bhanpuri, Nasir H; Okamura, Allison M; Bastian, Amy J

    2012-03-01

    Damage to the cerebellum causes characteristic movement abnormalities but is thought to have minimal impact on somatosensory perception. Traditional clinical assessments of patients with cerebellar lesions reveal no perceptual deficits despite the fact that the cerebellum receives substantial somatosensory information. Given that abnormalities have been reported in predicting the visual consequences of movement, we suspect that the cerebellum broadly participates in perception when motor output is required (i.e., active perception). Thus we hypothesize that cerebellar integrity is essential for somatosensory perception that requires motor activity, but not passive somatosensory perception. We compared the perceptual acuity of human cerebellar patients to that of healthy control subjects in several different somatosensory perception tasks with minimal visual information. We found that patients were worse at active force and stiffness discrimination but similar to control subjects with regard to passive cutaneous force detection, passive proprioceptive detection, and passive proprioceptive discrimination. Furthermore, the severity of movement symptoms as assessed by a clinical exam was positively correlated with impairment of active force perception. Notably, within the context of these perceptual tasks, control subjects and cerebellar patients displayed similar movement characteristics, and hence differing movement strategies are unlikely to underlie the differences in perception. Our results are consistent with the hypothesis that the cerebellum is vital to sensory prediction of self-generated movement and suggest a general role for the cerebellum in multiple forms of active perception.

  17. Binding profiles and physical dependence liabilities of selected benzodiazepine receptor ligands.

    PubMed

    Richards, J G; Martin, J R

    1998-01-01

    In vitro binding profiles were determined for selected benzodiazepine receptor (BZR) ligands by quantitative radioautography in rat brain. The ligands represent subtype-selective agonists (zolpidem) or nonselective BZR agonists (diazepam), as well as BZR partial agonists (bretazenil, Ro 43-9624, and Ro 19-8022). In addition, these compounds were evaluated in a precipitated withdrawal paradigm in monkeys. The physical dependence liability was not clearly related to the in vitro brain BZR binding profiles of these compounds. Therefore, diazepam, bretazenil, Ro 19-8022, and Ro 43-9624 had regional affinities for the 13 selected rat brain regions that were close to the mean values across regions, despite the clearly greater physical dependence potential of diazepam. Zolpidem, on the other hand, had regional affinities for the 13 rat brain regions that diverged significantly from the mean value across regions and exhibited a lower physical dependence potential than diazepam. These results raise the possibility that a combination of BZR subtype selectivity with partial agonism could yield a marked reduction of physical dependence liability.

  18. Large-Scale Profiling of Kinase Dependencies in Cancer Cell Lines

    PubMed Central

    Campbell, James; Ryan, Colm J.; Brough, Rachel; Bajrami, Ilirjana; Pemberton, Helen N.; Chong, Irene Y.; Costa-Cabral, Sara; Frankum, Jessica; Gulati, Aditi; Holme, Harriet; Miller, Rowan; Postel-Vinay, Sophie; Rafiq, Rumana; Wei, Wenbin; Williamson, Chris T.; Quigley, David A.; Tym, Joe; Al-Lazikani, Bissan; Fenton, Timothy; Natrajan, Rachael; Strauss, Sandra J.; Ashworth, Alan; Lord, Christopher J.

    2016-01-01

    Summary One approach to identifying cancer-specific vulnerabilities and therapeutic targets is to profile genetic dependencies in cancer cell lines. Here, we describe data from a series of siRNA screens that identify the kinase genetic dependencies in 117 cancer cell lines from ten cancer types. By integrating the siRNA screen data with molecular profiling data, including exome sequencing data, we show how vulnerabilities/genetic dependencies that are associated with mutations in specific cancer driver genes can be identified. By integrating additional data sets into this analysis, including protein-protein interaction data, we also demonstrate that the genetic dependencies associated with many cancer driver genes form dense connections on functional interaction networks. We demonstrate the utility of this resource by using it to predict the drug sensitivity of genetically or histologically defined subsets of tumor cell lines, including an increased sensitivity of osteosarcoma cell lines to FGFR inhibitors and SMAD4 mutant tumor cells to mitotic inhibitors. PMID:26947069

  19. Lipid Dependent Mechanisms of Protein Pump Activity

    DTIC Science & Technology

    1989-05-23

    properties which result form the colligative interactions of many lipid molecules. Important materials properties include . . . i I I II II I i I 1 the...d identify by olock number) *This project is aime at investigating if a lipid elastic property , known as the spontaneous radius of curvature Ro’, is...a regulated membrane property and if its value modulates membrane protein activity. Specific aims reported on here include: 1) Correlation of ion pump

  20. Power Dependence of the Electron Mobility Profile in a Hall Thruster

    NASA Technical Reports Server (NTRS)

    Jorns, Benjamin A.; Hofery, Richard H.; Mikellides, Ioannis G.

    2014-01-01

    The electron mobility profile is estimated in a 4.5 kW commercial Hall thruster as a function of discharge power. Internal measurements of plasma potential and electron temperature are made in the thruster channel with a high-speed translating probe. These measurements are presented for a range of throttling conditions from 150 - 400 V and 0.6 - 4.5 kW. The fluid-based solver, Hall2De, is used in conjunction with these internal plasma parameters to estimate the anomalous collision frequency profile at fixed voltage, 300 V, and three power levels. It is found that the anomalous collision frequency profile does not change significantly upstream of the location of the magnetic field peak but that the extent and magnitude of the anomalous collision frequency downstream of the magnetic peak does change with thruster power. These results are discussed in the context of developing phenomenological models for how the collision frequency profile depends on thruster operating conditions.

  1. Characteristic Changes in the Aroma Profile of Patchouli Depending on Manufacturing Process.

    PubMed

    Hasegawa, Toshio; Yoshitome, Kazuma; Fujihara, Takashi; Santoso, Mardi; Aziz, Muhammad Abdul

    2017-08-01

    Patchouli is used as an incense material and essential oil. The characteristic odor of patchouli leaves results from the drying process used in their production; however, there have to date been no reports on the changes in the odor of patchouli leaves during the drying process. We investigated the aroma profile of dried patchouli leaves using the hexane extracts of fresh and dried patchouli leaves. We focused on the presence or absence of the constituents of the fresh and dried extracts, and the differences in the content of the common constituents. Fourteen constituents were identified as characteristic of dried patchouli extract odor by gas chromatography-olfactometry analysis. The structures of seven of the 14 constituents were determined by gas chromatography-mass spectrometry (α-patchoulene, seychellene, humulene, α-bulnesene, isoaromadendrene epoxide, caryophyllene oxide, and patchouli alcohol). The aroma profile of the essential oil obtained from the dried patchouli leaves was clearly different from that of dried patchouli. The aroma profile of the essential oil was investigated by a similar method. We identified 12 compounds as important odor constituents. The structures of nine of the 12 constituents were determined by gas chromatography-mass spectrometry (cis-thujopsene, caryophyllene, α-guaiene, α-patchoulene, seychellene, α-bulnesene, isoaromadendrene epoxide, patchouli alcohol, and corymbolone). Comparing the odors and constituents demonstrated that the aroma profile of patchouli depends on the manufacturing process.

  2. Power Dependence of the Electron Mobility Profile in a Hall Thruster

    NASA Technical Reports Server (NTRS)

    Jorns, Benjamin A.; Hofery, Richard H.; Mikellides, Ioannis G.

    2014-01-01

    The electron mobility profile is estimated in a 4.5 kW commercial Hall thruster as a function of discharge power. Internal measurements of plasma potential and electron temperature are made in the thruster channel with a high-speed translating probe. These measurements are presented for a range of throttling conditions from 150 - 400 V and 0.6 - 4.5 kW. The fluid-based solver, Hall2De, is used in conjunction with these internal plasma parameters to estimate the anomalous collision frequency profile at fixed voltage, 300 V, and three power levels. It is found that the anomalous collision frequency profile does not change significantly upstream of the location of the magnetic field peak but that the extent and magnitude of the anomalous collision frequency downstream of the magnetic peak does change with thruster power. These results are discussed in the context of developing phenomenological models for how the collision frequency profile depends on thruster operating conditions.

  3. Range dependent sediment sound speed profile measurements using the image source method.

    PubMed

    Pinson, S; Guillon, L; Holland, C W

    2013-07-01

    This paper presents a range dependent sediment sound speed profile measurement obtained using the image source method. This technique is based on the analysis of the seafloor reflected acoustic wave as a collection of image sources which positions are linked with the thick-nesses and the sound speed of the sediment stack. The data used were acquired by the NURC in 2009 during the Clutter09 experiment. The equipment used was an autonomous undersea vehicle towing a 1600-3500 Hz frequency band source and a 32 m horizontal line array of 32 hydrophones at 12 m above the seabed. Under the assumption of locally range independent seabed properties, the moving horizontal array provides successive range independent sediment sound speed profiles along a track to obtain the range and depth dependent structure of the seafloor. Two key steps include recovery of the time-varying unknown array shape from the data and spatial filtering of the successive sound speed profiles. A comparison of the image source method result and seismic data along nearly the same 14 km track indicates that the seabed stratigraphy is correctly mapped by this method.

  4. Time-dependent solution of pre-mixed laminar flames with a known temperature profile

    SciTech Connect

    Olsson, J.O.; Andersson, L.L.

    1985-07-01

    A computer program designed for the evaluation of molecular flows interacting through chemical kinetics and molecular diffusion is described. Measured values of temperature profile and mass flow are used. The starting profiles and the hot boundary values are calculated by a kinetics approximation found by neglecting diffusion. A time-dependent method is used together with successive grid refinements. The successive grid refinements reduced the execution times by a factor of 5 for a H/sub 2//air flame at a pressure of 1 atm. For a CH/sub 4//O/sub 2/ flame at 0.05 atm the reduction due to grid refinements was a factor 50 or more according to the estimations. The execution times for the test flames were a factor 4 slower than a current implementation of the steady state method. Possible optimizations of the present time-dependent version can decrease that difference significantly. The computed concentration profiles agreed with published computed results with 1%.

  5. Order dependence of the profile of the intensities of multiple-quantum coherences

    SciTech Connect

    Lundin, A. A.; Zobov, V. E.

    2015-05-15

    A modification of the widespread phenomenological model theory of multiple-quantum (MQ) nuclear magnetic resonance spectra of a single cluster of correlated spins has been developed. In contrast to the mentioned theory, the size distribution of such clusters has been consistently taken into account. To obtain the distribution, solutions for the amplitudes of the expansion in the complete set of orthogonal operators are used. Expressions specifying the dependence of the profile of the intensities of MQ coherences on their number n (order) have been obtained. The total form of the dependence has been evaluated by means of the numerical implementation of the resulting expressions. The asymptotic expressions for large n values (wings of the spectrum) have been obtained analytically by the saddle-point method. It has been shown that the dependence under study has a Gaussian central part and exponential wings. The results obtained are in agreement with the previous calculations for some model systems and existing experimental data.

  6. A study of the properties of beryllium doped silicon with particular emphasis on diffusion mechanisms: Profiles of depth dependent conductivity as determined by electrical surface probes

    NASA Technical Reports Server (NTRS)

    Franks, R. K.; Robertson, J. B.

    1972-01-01

    Very large diffusion coefficients were encountered and required the determination of impurity profiles for samples approximately 1 cm thick. Since conductivity values are readily converted into concentrations of electrically active impurities, the major problem became that of accurately determining the conductivity profiles of beryllium diffused silicon samples. Four-point probe measurements on samples having depth conductivities are interpreted in terms of conductivity profiles, based on an exact solution of the problem of exponentially depth dependent conductivity. Applications include surface conductivity determination where the form of the conductivity profile is known, and conductivity profile determination from probe measurements taken as the sample surface is progressively lapped away. The application is limited to samples having conductivity monotonically decreasing with depth from the probed surface.

  7. Enzyme dynamics and activity: time-scale dependence of dynamical transitions in glutamate dehydrogenase solution.

    PubMed

    Daniel, R M; Finney, J L; Réat, V; Dunn, R; Ferrand, M; Smith, J C

    1999-10-01

    We have examined the temperature dependence of motions in a cryosolution of the enzyme glutamate dehydrogenase (GDH) and compared these with activity. Dynamic neutron scattering was performed with two instruments of different energy resolution, permitting the separate determination of the average dynamical mean square displacements on the sub-approximately 100 ps and sub-approximately 5 ns time scales. The results demonstrate a marked dependence on the time scale of the temperature profile of the mean square displacement. The lowest temperature at which anharmonic motion is observed is heavily dependent on the time window of the instrument used to observe the dynamics. Several dynamical transitions (inflexions of the mean squared displacement) are observed in the slower dynamics. Comparison with the temperature profile of the activity of the enzyme in the same solvent reveals dynamical transitions that have no effect on GDH function.

  8. Activity Dependent Signal Transduction in Skeletal Muscle

    NASA Technical Reports Server (NTRS)

    Hamilton, Susan L.

    1999-01-01

    The overall goals of this project are: 1) to define the initial signal transduction events whereby the removal of gravitational load from antigravity muscles, such as the soleus, triggers muscle atrophy, and 2) to develop countermeasures to prevent this from happening. Our rationale for this approach is that, if countermeasures can be developed to regulate these early events, we could avoid having to deal with the multiple cascades of events that occur downstream from the initial event. One of our major findings is that hind limb suspension causes an early and sustained increase in intracellular Ca(2+) concentration ([Ca (2+)](sub i)). In most cells the consequences of changes in ([Ca (2+)](sub i))depend on the amplitude, frequency and duration of the Ca(2+) signal and on other factors in the intracellular environment. We propose that muscle remodeling in microgravity represents a change in the balance among several CA(2+) regulated signal transduction pathways, in particular those involving the transcription factors NFAT and NFkB and the pro-apoptotic protein BAD. Other Ca(2+) sensitive pathways involving PKC, ras, rac, and CaM kinase II may also contribute to muscle remodeling.

  9. Activity Dependent Signal Transduction in Skeletal Muscle

    NASA Technical Reports Server (NTRS)

    Hamilton, Susan L.

    1999-01-01

    The overall goals of this project are: 1) to define the initial signal transduction events whereby the removal of gravitational load from antigravity muscles, such as the soleus, triggers muscle atrophy, and 2) to develop countermeasures to prevent this from happening. Our rationale for this approach is that, if countermeasures can be developed to regulate these early events, we could avoid having to deal with the multiple cascades of events that occur downstream from the initial event. One of our major findings is that hind limb suspension causes an early and sustained increase in intracellular Ca(2+) concentration ([Ca (2+)](sub i)). In most cells the consequences of changes in ([Ca (2+)](sub i))depend on the amplitude, frequency and duration of the Ca(2+) signal and on other factors in the intracellular environment. We propose that muscle remodeling in microgravity represents a change in the balance among several CA(2+) regulated signal transduction pathways, in particular those involving the transcription factors NFAT and NFkB and the pro-apoptotic protein BAD. Other Ca(2+) sensitive pathways involving PKC, ras, rac, and CaM kinase II may also contribute to muscle remodeling.

  10. Phenolic profile and antioxidant activity in selected seeds and sprouts.

    PubMed

    Pająk, Paulina; Socha, Robert; Gałkowska, Dorota; Rożnowski, Jacek; Fortuna, Teresa

    2014-01-15

    The aim of this study was to investigate the effect of germination on the phenolic acids and flavonoids profile, as well as antioxidant activity (AA), in selected edible seeds of mung beans, radish, broccoli and sunflower. Germination increased the total phenolic (TP) and flavonoid (TF) levels, as well as the AA of the seeds, and influenced the profile of free and bound phenolic compounds. Among the samples, mung bean was characterised by lowest levels of TP and TF, as well as AA, evaluated using ABTS, DPPH and FRAP assays. Sunflower and radish sprouts were the most rich in phenolic compounds. Insignificant amounts of free phenolic acids were found in the free phenolic acid fraction; alkaline hydrolysis of the seeds and sprouts extracts provided the majority of the phenolic acids. The amounts of free and bound flavonoids were inconsiderable both for seeds and sprouts. Copyright © 2013 Elsevier Ltd. All rights reserved.

  11. Universal Temporal Profile of Replication Origin Activation in Eukaryotes

    NASA Astrophysics Data System (ADS)

    Goldar, Arach

    2011-03-01

    The complete and faithful transmission of eukaryotic genome to daughter cells involves the timely duplication of mother cell's DNA. DNA replication starts at multiple chromosomal positions called replication origin. From each activated replication origin two replication forks progress in opposite direction and duplicate the mother cell's DNA. While it is widely accepted that in eukaryotic organisms replication origins are activated in a stochastic manner, little is known on the sources of the observed stochasticity. It is often associated to the population variability to enter S phase. We extract from a growing Saccharomyces cerevisiae population the average rate of origin activation in a single cell by combining single molecule measurements and a numerical deconvolution technique. We show that the temporal profile of the rate of origin activation in a single cell is similar to the one extracted from a replicating cell population. Taking into account this observation we exclude the population variability as the origin of observed stochasticity in origin activation. We confirm that the rate of origin activation increases in the early stage of S phase and decreases at the latter stage. The population average activation rate extracted from single molecule analysis is in prefect accordance with the activation rate extracted from published micro-array data, confirming therefore the homogeneity and genome scale invariance of dynamic of replication process. All these observations point toward a possible role of replication fork to control the rate of origin activation.

  12. Age-dependent variation of the Gradient Index profile in human crystalline lenses

    PubMed Central

    de Castro, A.; Siedlecki, D.; Borja, David; Uhlhorn, Stephen; Parel, Jean-Marie; Manns, Fabrice; Marcos, S.

    2011-01-01

    Purpose To reconstruct the gradient index (GRIN) profile of human crystalline lenses ex-vivo using Optical Coherence Tomography (OCT) imaging with an optimization technique and to study the dependence of the GRIN profile with age. Methods Cross-sectional images of nine isolated human crystalline lenses with ages ranging from 6 to 72 (post mortem time 1 to 4 days) were obtained using a custom-made OCT system. Lenses were extracted from whole cadaver globes and placed in a measurement chamber filled with preservation medium (DMEM). Lenses were imaged with the anterior surface up and then flipped over and imaged again, to obtain posterior lens surface profiles both undistorted and distorted by the refraction through the anterior crystalline lens and GRIN. The GRIN distribution of the lens was described with three variables by means of power function, with variables being the nucleus and surface index, and a power coefficient that describes the decay of the refractive index from the nucleus to the surface. An optimization method was used to search for the parameters that produced the best match of the distorted posterior surface. Results The distorted surface was simulated with accuracy around the resolution of the OCT system (under 15 µm). The reconstructed refractive index values ranged from 1.356 to 1.388 for the surface, and from 1.396 to 1.434 for the nucleus. The power coefficient ranged between 3 and 18. The power coefficient increased significantly with age, at a rate of 0.24 per year. Conclusion Optical Coherence Tomography allowed optical, non-invasive measurement of the 2-D gradient index profile of the isolated human crystalline lens ex vivo. The age-dependent variation of the changes is consistent with previous data using magnetic resonance imaging, and the progressive formation of a refractive index plateau. PMID:22865954

  13. Age-dependent variation of the gradient index profile in human crystalline lenses

    NASA Astrophysics Data System (ADS)

    de Castro, Alberto; Siedlecki, Damian; Borja, David; Uhlhorn, Stephen; Parel, Jean-Marie; Manns, Fabrice; Marcos, Susana

    2011-11-01

    An investigation was carried out with the aim of reconstructing the gradient index (GRIN) profile of human crystalline lenses ex-vivo using optical coherence tomography (OCT) imaging with an optimization technique and to study the dependence of the GRIN profile with age. Cross-sectional images of nine isolated human crystalline lenses with ages ranging from 6 to 72 (post-mortem time 1 to 4 days) were obtained using a custom-made OCT system. Lenses were extracted from whole cadaver globes and placed in a measurement chamber filled with preservation medium (DMEM). Lenses were imaged with the anterior surface up and then flipped over and imaged again, to obtain posterior lens surface profiles both undistorted and distorted by the refraction through the anterior crystalline lens and GRIN. The GRIN distribution of the lens was described with three variables by means of power function, with variables being the nucleus and surface index, and a power coefficient that describes the decay of the refractive index from the nucleus to the surface. An optimization method was used to search for the parameters that produced the best match of the distorted posterior surface. The distorted surface was simulated with accuracy around the resolution of the OCT system (under 15 µm). The reconstructed refractive index values ranged from 1.356 to 1.388 for the surface, and from 1.396 to 1.434 for the nucleus. The power coefficient ranged between 3 and 18. The power coefficient increased significantly with age, at a rate of 0.24 per year. Optical coherence tomography allowed optical, non-invasive measurement of the 2D gradient index profile of the isolated human crystalline lens ex vivo. The age-dependent variation of the changes is consistent with previous data using magnetic resonance imaging, and the progressive formation of a refractive index plateau.

  14. Strings on a Violin: Location Dependence of Frequency Tuning in Active Dendrites.

    PubMed

    Das, Anindita; Rathour, Rahul K; Narayanan, Rishikesh

    2017-01-01

    Strings on a violin are tuned to generate distinct sound frequencies in a manner that is firmly dependent on finger location along the fingerboard. Sound frequencies emerging from different violins could be very different based on their architecture, the nature of strings and their tuning. Analogously, active neuronal dendrites, dendrites endowed with active channel conductances, are tuned to distinct input frequencies in a manner that is dependent on the dendritic location of the synaptic inputs. Further, disparate channel expression profiles and differences in morphological characteristics could result in dendrites on different neurons of the same subtype tuned to distinct frequency ranges. Alternately, similar location-dependence along dendritic structures could be achieved through disparate combinations of channel profiles and morphological characteristics, leading to degeneracy in active dendritic spectral tuning. Akin to strings on a violin being tuned to different frequencies than those on a viola or a cello, different neuronal subtypes exhibit distinct channel profiles and disparate morphological characteristics endowing each neuronal subtype with unique location-dependent frequency selectivity. Finally, similar to the tunability of musical instruments to elicit distinct location-dependent sounds, neuronal frequency selectivity and its location-dependence are tunable through activity-dependent plasticity of ion channels and morphology. In this morceau, we explore the origins of neuronal frequency selectivity, and survey the literature on the mechanisms behind the emergence of location-dependence in distinct forms of frequency tuning. As a coda to this composition, we present some future directions for this exciting convergence of biophysical mechanisms that endow a neuron with frequency multiplexing capabilities.

  15. Strings on a Violin: Location Dependence of Frequency Tuning in Active Dendrites

    PubMed Central

    Das, Anindita; Rathour, Rahul K.; Narayanan, Rishikesh

    2017-01-01

    Strings on a violin are tuned to generate distinct sound frequencies in a manner that is firmly dependent on finger location along the fingerboard. Sound frequencies emerging from different violins could be very different based on their architecture, the nature of strings and their tuning. Analogously, active neuronal dendrites, dendrites endowed with active channel conductances, are tuned to distinct input frequencies in a manner that is dependent on the dendritic location of the synaptic inputs. Further, disparate channel expression profiles and differences in morphological characteristics could result in dendrites on different neurons of the same subtype tuned to distinct frequency ranges. Alternately, similar location-dependence along dendritic structures could be achieved through disparate combinations of channel profiles and morphological characteristics, leading to degeneracy in active dendritic spectral tuning. Akin to strings on a violin being tuned to different frequencies than those on a viola or a cello, different neuronal subtypes exhibit distinct channel profiles and disparate morphological characteristics endowing each neuronal subtype with unique location-dependent frequency selectivity. Finally, similar to the tunability of musical instruments to elicit distinct location-dependent sounds, neuronal frequency selectivity and its location-dependence are tunable through activity-dependent plasticity of ion channels and morphology. In this morceau, we explore the origins of neuronal frequency selectivity, and survey the literature on the mechanisms behind the emergence of location-dependence in distinct forms of frequency tuning. As a coda to this composition, we present some future directions for this exciting convergence of biophysical mechanisms that endow a neuron with frequency multiplexing capabilities. PMID:28348519

  16. Universal Temporal Profile of Replication Origin Activation in Eukaryotes

    PubMed Central

    Goldar, Arach; Marsolier-Kergoat, Marie-Claude; Hyrien, Olivier

    2009-01-01

    Although replication proteins are conserved among eukaryotes, the sequence requirements for replication initiation differ between species. In all species, however, replication origins fire asynchronously throughout S phase. The temporal program of origin firing is reproducible in cell populations but largely probabilistic at the single-cell level. The mechanisms and the significance of this program are unclear. Replication timing has been correlated with gene activity in metazoans but not in yeast. One potential role for a temporal regulation of origin firing is to minimize fluctuations in replication end time and avoid persistence of unreplicated DNA in mitosis. Here, we have extracted the population-averaged temporal profiles of replication initiation rates for S. cerevisiae, S. pombe, D. melanogaster, X. laevis and H. sapiens from genome-wide replication timing and DNA combing data. All the profiles have a strikingly similar shape, increasing during the first half of S phase then decreasing before its end. A previously proposed minimal model of stochastic initiation modulated by accumulation of a recyclable, limiting replication-fork factor and fork-promoted initiation of new origins, quantitatively described the observed profiles without requiring new implementations. The selective pressure for timely completion of genome replication and optimal usage of replication proteins that must be imported into the cell nucleus can explain the generic shape of the profiles. We have identified a universal behavior of eukaryotic replication initiation that transcends the mechanisms of origin specification. The population-averaged efficiency of replication origin usage changes during S phase in a strikingly similar manner in a highly diverse set of eukaryotes. The quantitative model previously proposed for origin activation in X. laevis can be generalized to explain this evolutionary conservation. PMID:19521533

  17. Transcriptome profiling reveals the regulatory mechanism underlying pollination dependent and parthenocarpic fruit set mainly mediated by auxin and gibberellin.

    PubMed

    Tang, Ning; Deng, Wei; Hu, Guojian; Hu, Nan; Li, Zhengguo

    2015-01-01

    Fruit set is a key process for crop production in tomato which occurs after successful pollination and fertilization naturally. However, parthenocarpic fruit development can be uncoupled from fertilization triggered by exogenous auxin or gibberellins (GAs). Global transcriptome knowledge during fruit initiation would help to characterize the molecular mechanisms by which these two hormones regulate pollination-dependent and -independent fruit set. In this work, digital gene expression tag profiling (DGE) technology was applied to compare the transcriptomes from pollinated and 2, 4-D/GA3-treated ovaries. Activation of carbohydrate metabolism, cell division and expansion as well as the down-regulation of MADS-box is a comprehensive regulatory pathway during pollination-dependent and parthenocarpic fruit set. The signaling cascades of auxin and GA are significantly modulated. The feedback regulations of Aux/IAAs and DELLA genes which functioned to fine-tune auxin and GA response respectively play fundamental roles in triggering fruit initiation. In addition, auxin regulates GA synthesis via up-regulation of GA20ox1 and down-regulation of KNOX. Accordingly, the effect of auxin on fruit set is mediated by GA via ARF2 and IAA9 down-regulation, suggesting that both pollination-dependent and parthenocarpic fruit set depend on the crosstalk between auxin and GA. This study characterizes the transcriptomic features of ovary development and more importantly unravels the integral roles of auxin and GA on pollination-dependent and parthenocarpic fruit set.

  18. Transcriptome Profiling Reveals the Regulatory Mechanism Underlying Pollination Dependent and Parthenocarpic Fruit Set Mainly Mediated by Auxin and Gibberellin

    PubMed Central

    Tang, Ning; Deng, Wei; Hu, Guojian; Hu, Nan; Li, Zhengguo

    2015-01-01

    Background Fruit set is a key process for crop production in tomato which occurs after successful pollination and fertilization naturally. However, parthenocarpic fruit development can be uncoupled from fertilization triggered by exogenous auxin or gibberellins (GAs). Global transcriptome knowledge during fruit initiation would help to characterize the molecular mechanisms by which these two hormones regulate pollination-dependent and -independent fruit set. Principal Findings In this work, digital gene expression tag profiling (DGE) technology was applied to compare the transcriptomes from pollinated and 2, 4-D/GA3-treated ovaries. Activation of carbohydrate metabolism, cell division and expansion as well as the down-regulation of MADS-box is a comprehensive regulatory pathway during pollination-dependent and parthenocarpic fruit set. The signaling cascades of auxin and GA are significantly modulated. The feedback regulations of Aux/IAAs and DELLA genes which functioned to fine-tune auxin and GA response respectively play fundamental roles in triggering fruit initiation. In addition, auxin regulates GA synthesis via up-regulation of GA20ox1 and down-regulation of KNOX. Accordingly, the effect of auxin on fruit set is mediated by GA via ARF2 and IAA9 down-regulation, suggesting that both pollination-dependent and parthenocarpic fruit set depend on the crosstalk between auxin and GA. Significance This study characterizes the transcriptomic features of ovary development and more importantly unravels the integral roles of auxin and GA on pollination-dependent and parthenocarpic fruit set. PMID:25909657

  19. Biological spectra analysis: Linking biological activity profiles to molecular structure

    PubMed Central

    Fliri, Anton F.; Loging, William T.; Thadeio, Peter F.; Volkmann, Robert A.

    2005-01-01

    Establishing quantitative relationships between molecular structure and broad biological effects has been a longstanding challenge in science. Currently, no method exists for forecasting broad biological activity profiles of medicinal agents even within narrow boundaries of structurally similar molecules. Starting from the premise that biological activity results from the capacity of small organic molecules to modulate the activity of the proteome, we set out to investigate whether descriptor sets could be developed for measuring and quantifying this molecular property. Using a 1,567-compound database, we show that percent inhibition values, determined at single high drug concentration in a battery of in vitro assays representing a cross section of the proteome, provide precise molecular property descriptors that identify the structure of molecules. When broad biological activity of molecules is represented in spectra form, organic molecules can be sorted by quantifying differences between biological spectra. Unlike traditional structure–activity relationship methods, sorting of molecules by using biospectra comparisons does not require knowledge of a molecule's putative drug targets. To illustrate this finding, we selected as starting point the biological activity spectra of clotrimazole and tioconazole because their putative target, lanosterol demethylase (CYP51), was not included in the bioassay array. Spectra similarity obtained through profile similarity measurements and hierarchical clustering provided an unbiased means for establishing quantitative relationships between chemical structures and biological activity spectra. This methodology, which we have termed biological spectra analysis, provides the capability not only of sorting molecules on the basis of biospectra similarity but also of predicting simultaneous interactions of new molecules with multiple proteins. PMID:15625110

  20. Interaction of silver nanoparticles with proteins: a characteristic protein concentration dependent profile of SPR signal.

    PubMed

    Banerjee, Victor; Das, K P

    2013-11-01

    Silver nanoparticles are finding increasing applications in biological systems, for example as antimicrobial agents and potential candidates for control drug release systems. In all such applications, silver nanoparticles interact with proteins and other biomolecules. Hence, the study of such interactions is of considerable importance. While BSA has been extensively used as a model protein for the study of interaction with the silver nanoparticles, studies using other proteins are rather limited. The interaction of silver nanoparticles with light leads to collective oscillation of the conducting electrons giving rise to surface plasmon resonance (SPR). Here, we have studied the protein concentration dependence of the SPR band profiles for a number of proteins. We found that for all the proteins, with increase in concentration, the SPR band intensity initially decreased, reaching minima and then increased again leading to a characteristic "dip and rise" pattern. Minimum point of the pattern appeared to be related to the isoelectric point of the proteins. Detailed dynamic light scattering and transmission electron microscopy studies revealed that the consistency of SPR profile was dependent on the average particle size and state of association of the silver nanoparticles with the change in the protein concentration. Fluorescence spectroscopic studies showed the binding constants of the proteins with the silver nanoparticles were in the nano molar range with more than one nanoparticle binding to protein molecule. Structural studies demonstrate that protein retains its native-like structure on the nanoparticle surface unless the molar ratio of silver nanoparticles to protein exceeds 10. Our study reveals that nature of the protein concentration dependent profile of SPR signal is a general phenomena and mostly independent of the size and structure of the proteins. Copyright © 2013 Elsevier B.V. All rights reserved.

  1. Size Dependent Antioxidant Activity of Polypyrrole Nanofibers

    NASA Astrophysics Data System (ADS)

    Kumar, A.; Banerjee, Somik

    2011-07-01

    Polypyrrole (PPy) nanofibers have been synthesized employing surfactant assisted miceller polymerization by varying the surfactant concentration. The synthesized nanofibers have been characterized using TEM, XRD, FTIR and UV-Visible spectroscopy. TEM reveals that the diameter of the PPy nanowires decreases with the increase in surfactant concentration. X-ray spectra shows an amorphous peak centered around 2θ = 24.6° which is attributed to the π-π interaction of the partial PPy chains similar to that of aromatic groups. The domain length of the samples determined using the single-line approximation technique, decreases with decreasing diameter whereas the strain in the material increases, which have been attributed to the reduction of size with increase in the surfactant concentration as revealed by TEM. The vibrational bands observed from the FTIR spectra confirm the formation of surfactant free PPy nanowires. UV-Visible spectra shows a blue-shift in the π-π* absorption peak. Antioxidant activity of the samples has been determined using the DPPH free radical method. It has been observed that enhancement in free radical scavenging coincides with the decreasing diameter of the PPy nanofibers which has been associated with the increase in the surface reaction sites with the reduction of size.

  2. Voltage Dependence of a Neuromodulator-Activated Ionic Current.

    PubMed

    Gray, Michael; Golowasch, Jorge

    2016-01-01

    The neuromodulatory inward current (IMI) generated by crab Cancer borealis stomatogastric ganglion neurons is an inward current whose voltage dependence has been shown to be crucial in the activation of oscillatory activity of the pyloric network of this system. It has been previously shown that IMI loses its voltage dependence in conditions of low extracellular calcium, but that this effect appears to be regulated by intracellular calmodulin. Voltage dependence is only rarely regulated by intracellular signaling mechanisms. Here we address the hypothesis that the voltage dependence of IMI is mediated by intracellular signaling pathways activated by extracellular calcium. We demonstrate that calmodulin inhibitors and a ryanodine antagonist can reduce IMI voltage dependence in normal Ca(2+), but that, in conditions of low Ca(2+), calmodulin activators do not restore IMI voltage dependence. Further, we show evidence that CaMKII alters IMI voltage dependence. These results suggest that calmodulin is necessary but not sufficient for IMI voltage dependence. We therefore hypothesize that the Ca(2+)/calmodulin requirement for IMI voltage dependence is due to an active sensing of extracellular calcium by a GPCR family calcium-sensing receptor (CaSR) and that the reduction in IMI voltage dependence by a calmodulin inhibitor is due to CaSR endocytosis. Supporting this, preincubation with an endocytosis inhibitor prevented W7 (N-(6-aminohexyl)-5-chloro-1-naphthalenesulfonamide hydrochloride)-induced loss of IMI voltage dependence, and a CaSR antagonist reduced IMI voltage dependence. Additionally, myosin light chain kinase, which is known to act downstream of the CaSR, seems to play a role in regulating IMI voltage dependence. Finally, a Gβγ-subunit inhibitor also affects IMI voltage dependence, in support of the hypothesis that this process is regulated by a G-protein-coupled CaSR.

  3. Voltage Dependence of a Neuromodulator-Activated Ionic Current123

    PubMed Central

    2016-01-01

    Abstract The neuromodulatory inward current (IMI) generated by crab Cancer borealis stomatogastric ganglion neurons is an inward current whose voltage dependence has been shown to be crucial in the activation of oscillatory activity of the pyloric network of this system. It has been previously shown that IMI loses its voltage dependence in conditions of low extracellular calcium, but that this effect appears to be regulated by intracellular calmodulin. Voltage dependence is only rarely regulated by intracellular signaling mechanisms. Here we address the hypothesis that the voltage dependence of IMI is mediated by intracellular signaling pathways activated by extracellular calcium. We demonstrate that calmodulin inhibitors and a ryanodine antagonist can reduce IMI voltage dependence in normal Ca2+, but that, in conditions of low Ca2+, calmodulin activators do not restore IMI voltage dependence. Further, we show evidence that CaMKII alters IMI voltage dependence. These results suggest that calmodulin is necessary but not sufficient for IMI voltage dependence. We therefore hypothesize that the Ca2+/calmodulin requirement for IMI voltage dependence is due to an active sensing of extracellular calcium by a GPCR family calcium-sensing receptor (CaSR) and that the reduction in IMI voltage dependence by a calmodulin inhibitor is due to CaSR endocytosis. Supporting this, preincubation with an endocytosis inhibitor prevented W7 (N-(6-aminohexyl)-5-chloro-1-naphthalenesulfonamide hydrochloride)-induced loss of IMI voltage dependence, and a CaSR antagonist reduced IMI voltage dependence. Additionally, myosin light chain kinase, which is known to act downstream of the CaSR, seems to play a role in regulating IMI voltage dependence. Finally, a Gβγ-subunit inhibitor also affects IMI voltage dependence, in support of the hypothesis that this process is regulated by a G-protein-coupled CaSR. PMID:27257619

  4. Profile of internet access in active cocaine users.

    PubMed

    Strickland, Justin C; Wagner, Frances P; Stoops, William W; Rush, Craig R

    2015-10-01

    Web-based interventions have received attention for substance abuse treatment. Few studies have examined Internet use among substance users. Internet-use data were examined for 66 participants screened to participate in behavioral pharmacology studies. A majority of active cocaine users reported regular Internet use. Demographic profiles generally did not impact Internet use, but Internet users were more likely to be younger and report other drug use. Active cocaine users have similar rates of Internet access as the general population. Our findings contribute to the limited data on Internet use in active drug users by demonstrating Internet access in cocaine-using populations, supporting the use of this medium to conduct research and clinical interventions. © American Academy of Addiction Psychiatry.

  5. Calcium Activation Profile In Electrically Stimulated Intact Rat Heart Cells

    NASA Astrophysics Data System (ADS)

    Geerts, Hugo; Nuydens, Rony; Ver Donck, Luc; Nuyens, Roger; De Brabander, Marc; Borgers, Marcel

    1988-06-01

    Recent advances in fluorescent probe technology and image processing equipment have made available the measurement of calcium in living systems on a real-time basis. We present the use of the calcium indicator Fura-2 in intact normally stimulated rat heart cells for the spatial and dynamic measurement of the calcium excitation profile. After electric stimulation (1 Hz), the activation proceeds from the center of the myocyte toward the periphery. Within two frame times (80 ms), the whole cell is activated. The activation is slightly faster in the center of the cell than in the periphery. The mean recovery time is 200-400 ms. There is no difference along the cell's long axis. The effect of a beta-agonist and of a calcium antagonist is described.

  6. Phytochemical profiles and antioxidant activity of processed brown rice products.

    PubMed

    Gong, Er Sheng; Luo, Shunjing; Li, Tong; Liu, Chengmei; Zhang, Guowen; Chen, Jun; Zeng, Zicong; Liu, Rui Hai

    2017-10-01

    The phytochemical profiles and antioxidant activity of free, soluble-conjugated, and bound fractions of brown rice and its processed products (textured rice, cooked rice and rice noodle) were studied. Nineteen phenolic acids were identified. Trans-ferulic acid was the most abundant monomeric phenolic acid with trans-trans-8-O-4' diferulic acid being most abundant diferulic acid. Processing increased the content of free phenolic acids, but decreased the content of soluble-conjugated phenolic acids. The content of bound phenolic acids was increased by improved extrusion cooking technology and cooking, but not affected by rice noodle extrusion. The total phenolic contents and antioxidant activities of free and soluble-conjugated fractions were decreased after processing, whereas those of bound fraction were increased by improved extrusion cooking technology and cooking, but not affected by rice noodle extrusion. Results indicated that whole foods designed for reducing chronic disease risk need to consider the effects of processing on phytochemical profiles and antioxidant activity of whole grains. Copyright © 2017 Elsevier Ltd. All rights reserved.

  7. Chemical profiling and antioxidant activity of Bolivian propolis.

    PubMed

    Nina, Nélida; Quispe, Cristina; Jiménez-Aspee, Felipe; Theoduloz, Cristina; Giménez, Alberto; Schmeda-Hirschmann, Guillermo

    2016-04-01

    Propolis is a relevant research subject worldwide. However, there is no information so far on Bolivian propolis. Ten propolis samples were collected from regions with high biodiversity in the main honey production places in Bolivia and were analyzed for their total phenolics (TP), flavonoids (TF) and antioxidant activity. The chemical profiles of the samples were assessed by TLC, HPLC-DAD, HPLC-DAD-MS/MS(n) and NMR analysis. TP, TF, TLC and NMR analysis showed significant chemical differences between the samples. Isolation of the main constituents by chromatography and identification by HPLC-DAD-MS/MS(n) achieved more than 35 constituents. According to their profiles, the Bolivian propolis can be classified into phenolic-rich and triterpene-rich samples. Propolis from the valleys (Cochabamba, Chuquisaca and Tarija) contained mainly prenylated phenylpropanoids, while samples from La Paz and Santa Cruz contained cycloartane and pentacyclic triterpenes. Phenolic-rich samples presented moderate to strong antioxidant activity while the triterpene-rich propolis were weakly active. High chemical diversity and differential antioxidant effects were found in Bolivian propolis. Our results provide additional evidence on the chemical composition and bioactivity of South American propolis. © 2015 Society of Chemical Industry.

  8. Kinase activity profiling of gram-negative pneumonia.

    PubMed

    Hoogendijk, Arie J; Diks, Sander H; Peppelenbosch, Maikel P; Van Der Poll, Tom; Wieland, Catharina W

    2011-01-01

    Pneumonia is a severe disease with high morbidity and mortality. A major causative pathogen is the Gram-negative bacterium Klebsiella (K.) pneumoniae. Kinases play an integral role in the transduction of intracellular signaling cascades and regulate a diverse array of biological processes essential to immune cells. The current study explored signal transduction events during murine Gram-negative pneumonia using a systems biology approach. Kinase activity arrays enable the analysis of 1,024 consensus sequences of protein kinase substrates. Using a kinase activity array on whole lung lysates, cellular kinase activities were determined in a mouse model of K. pneumoniae pneumonia. Notable kinase activities also were validated with phospho-specific Western blots. On the basis of the profiling data, mitogen-activated protein kinase (MAPK) signaling via p42 mitogen-activated protein kinase (p42) and p38 mitogen-activated protein kinase (p38) and transforming growth factor β (TGFβ) activity were reduced during infection, whereas v-src sarcoma (Schmidt-Ruppin A-2) viral oncogene homolog (avian) (SRC) activity generally was enhanced. AKT signaling was represented in both metabolic and inflammatory (mitogen-activated protein kinase kinase 2 [MKK], apoptosis signal-regulating kinase/mitogen-activated protein kinase kinase kinase 5 [ASK] and v-raf murine sarcoma viral oncogene homolog B1 [b-RAF]) context. This study reaffirms the importance of classic inflammation pathways, such as MAPK and TGFβ signaling and reveals less known involvement of glycogen synthase kinase 3β (GSK-3β), AKT and SRC signaling cassettes in pneumonia.

  9. Dose dependence of surface damage profiles for Ge(111) irradiated with 3 keV Ar +

    NASA Astrophysics Data System (ADS)

    Kido, Yoshiaki; Nakano, Hirohiko

    1990-12-01

    The damage profiles of 3 keV Ar + -irradiated Ge(111) were measured by medium energy ion scattering (MEIS) with 220 keV H + beams. A depth resolution of 0.5 nm has been achieved using an electrostatic toroidal analyzer. With doses over 1 × 10 14Ar+/ cm2 at room temperature, an amorphous layer is formed and the amorphous layer thickness is saturated by a dose of 1 × 10 15Ar+/ cm2. The saturated amorphous layer thickness of 12-13 nm obtained is in good agreement with that observed by a cross section transmission electron microscope. The dose dependence of the induced defects derived from MEIS is consistent with that determined by electron channeling pattern analysis. The amorphized damage profiles determined directly by MEIS agree well with those obtained by solving the rate equation using the unsaturated initial defect profile for a dose of 2 × 10 13Ar+/ cm2 and assuming a sputtering rate of 2.4-3.0.

  10. Examining the profile of high-potency cannabis and its association with severity of cannabis dependence.

    PubMed

    Freeman, T P; Winstock, A R

    2015-11-01

    Cannabis use is decreasing in England and Wales, while demand for cannabis treatment in addiction services continues to rise. This could be partly due to an increased availability of high-potency cannabis. Adults residing in the UK were questioned about their drug use, including three types of cannabis (high potency: skunk; low potency: other grass, resin). Cannabis types were profiled and examined for possible associations between frequency of use and (i) cannabis dependence, (ii) cannabis-related concerns. Frequent use of high-potency cannabis predicted a greater severity of dependence [days of skunk use per month: b = 0.254, 95% confidence interval (CI) 0.161-0.357, p < 0.001] and this effect became stronger as age decreased (b = -0.006, 95% CI -0.010 to -0.002, p = 0.004). By contrast, use of low-potency cannabis was not associated with dependence (days of other grass use per month: b = 0.020, 95% CI -0.029 to 0.070, p = 0.436; days of resin use per month: b = 0.025, 95% CI -0.019 to 0.067, p = 0.245). Frequency of cannabis use (all types) did not predict severity of cannabis-related concerns. High-potency cannabis was clearly distinct from low-potency varieties by its marked effects on memory and paranoia. It also produced the best high, was preferred, and most available. High-potency cannabis use is associated with an increased severity of dependence, especially in young people. Its profile is strongly defined by negative effects (memory, paranoia), but also positive characteristics (best high, preferred type), which may be important when considering clinical or public health interventions focusing on cannabis potency.

  11. A generalizability study of the instrumental activities of daily living profile.

    PubMed

    Bottari, Carolina; Dassa, Clément; Rainville, Constant; Dutil, Elisabeth

    2010-05-01

    To establish generalizability estimates of the Instrumental Activities of Daily Living (IADL) Profile when administered to persons with a moderate or severe traumatic brain injury (TBI) within their home and community environment. Generalizability theory was used to estimate generalizability and dependability coefficients as well as the relative contribution of identified sources of measurement error to total measurement error. Decision studies were used to enable the investigators to determine the optimal measurement design. The IADL Profile was administered in subjects' homes and community environments. Convenience sample of adults with a moderate or severe TBI (N=30, aged 16-65y). Not applicable. IADL Profile scores include 6 factor scores (going to the grocery store/shopping for groceries, having a meal with guests/cleaning up, putting on outdoor clothing, obtaining information, making a budget, and preparing a hot meal) and 1 total score. The greatest sources of measurement error were the subject-item interactions (3-random-facet design) and the subject-rater interactions (2-random-facet design). One hundred percent of generalizability coefficients of factor scores indicated acceptable to excellent reliability. Indices of dependability confirmed that 1 evaluator could reliably score the tool on a single occasion after having received a 3-day training workshop. The IADL Profile administered to persons with a moderate or severe TBI provides occupational therapists with a reliable set of measures of IADL independence capable of both capturing and analyzing the complex interactions between personal and environmental factors.

  12. [Physical activity during cancer: Can we define participants' profiles?

    PubMed

    Villaron, Charlène; Marqueste, Tanguy; Eisinger, François; Cappiello, Maria Antonietta; Cury, François

    2017-03-01

    Benefits of physical activity during cancer treatment are widely demonstrated, however, most of patients are not active enough. Several studies have analyzed the different variables that would affect the participation to physical activity programs. The aim of our study was to define profiles of patients who agree to participate in a physical activity program in the medical setting according to the hospital structure in which they receive their care, their past and present habits in sports and their temporal perspectives. Forty-six patients treated from two different hospitals (regional hospital denoted CLCC; and local hospital denoted CH), completed a survey consisting of a questionnaire on their past and present habits in physical activity, ZTPI and a demographic questionnaire. Patients could decide to participate or not in a physical activity program in the medical community. T-tests and Chi(2) were performed to compare the two groups. Chi(2) tests have shown that patients cared in CH are significantly more involved in physical activity program than patients cared in CLCC. Our study points out that the past and present patient PA (physical activity) has no influence on their accession to a physical activity program, however the type of hospital providing patient care could influence their participation. These results should lead us to rethink about the different forms of communication made around the physical activity programs in medical contexts, and about different practical arrangements proposed according to each health facility. Copyright © 2016 Société Française du Cancer. Published by Elsevier Masson SAS. All rights reserved.

  13. Profiles.

    ERIC Educational Resources Information Center

    Macintosh, Henry G.

    An introduction to profiles is presented with examples provided to permit an overall appraisal of the potential of profiles, of the principles upon which they might be based, and of the problems that will have to be overcome if their potential is to be realized in practice. The larger scale examples of profiles discussed are the Scottish Pupil…

  14. Monitoring of Time-Dependent System Profiles by Multiplex Gas Chromatography with Maximum Entropy Demodulation

    NASA Technical Reports Server (NTRS)

    Becker, Joseph F.; Valentin, Jose

    1996-01-01

    The maximum entropy technique was successfully applied to the deconvolution of overlapped chromatographic peaks. An algorithm was written in which the chromatogram was represented as a vector of sample concentrations multiplied by a peak shape matrix. Simulation results demonstrated that there is a trade off between the detector noise and peak resolution in the sense that an increase of the noise level reduced the peak separation that could be recovered by the maximum entropy method. Real data originated from a sample storage column was also deconvoluted using maximum entropy. Deconvolution is useful in this type of system because the conservation of time dependent profiles depends on the band spreading processes in the chromatographic column, which might smooth out the finer details in the concentration profile. The method was also applied to the deconvolution of previously interpretted Pioneer Venus chromatograms. It was found in this case that the correct choice of peak shape function was critical to the sensitivity of maximum entropy in the reconstruction of these chromatograms.

  15. Optimality of a time-dependent treatment profile during an epidemic

    PubMed Central

    Jaberi-Douraki, Majid; Moghadas, Seyed M.

    2013-01-01

    The emergence and spread of drug resistance is one of the most challenging public health issues in the treatment of some infectious diseases. The objective of this work is to investigate whether the effect of resistance can be contained through a time-dependent treatment strategy during the epidemic subject to an isoperimetric constraint. We apply control theory to a population dynamical model of influenza infection with drug-sensitive and drug-resistant strains, and solve the associated control problem to find the optimal treatment profile that minimizes the cumulative number of infections (i.e. the epidemic final size). We consider the problem under the assumption of limited drug stockpile and show that as the size of stockpile increases, a longer delay in start of treatment is required to minimize the total number of infections. Our findings show that the amount of drugs used to minimize the total number of infections depends on the rate of de novo resistance regardless of the initial size of drug stockpile. We demonstrate that both the rate of resistance emergence and the relative transmissibility of the resistant strain play important roles in determining the optimal timing and level of treatment profile. AMS Subject Classification: 34, 49, 92 PMID:23859002

  16. Intracellular CHO Cell Metabolite Profiling Reveals Steady-State Dependent Metabolic Fingerprints in Perfusion Culture.

    PubMed

    Karst, Daniel J; Steinhoff, Robert F; Kopp, Marie R G; Serra, Elisa; Soos, Miroslav; Zenobi, Renato; Morbidelli, Massimo

    2016-12-20

    Perfusion cell culture processes allow the steady-state culture of mammalian cells at high viable cell density, which is beneficial for overall product yields and homogeneity of product quality in the manufacturing of therapeutic proteins. In this study, the extent of metabolic steady state and the change of the metabolite profile between different steady states of an industrial Chinese hamster ovary (CHO) cell line producing a monoclonal antibody (mAb) was investigated in stirred tank perfusion bioreactors. Matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) of daily cell extracts revealed more than a hundred peaks, among which 76 metabolites were identified by tandem MS (MS/MS) and high resolution Fourier transform ion cyclotron resonance (FT-ICR) MS. Nucleotide ratios (Uridine (U)-ratio, nucleotide triphosphate (NTP)-ratio and energy charge (EC)) and multivariate analysis of all features indicated a consistent metabolite profile for a stable culture performed at 40 × 10(6) cells/mL over 26 days of culture. Conversely, the reactor was operated continuously so as to reach three distinct steady states one after the other at 20, 60, and 40 × 10(6) cells/mL. In each case, a stable metabolite profile was achieved after an initial transient phase of approximately three days at constant cell density when varying between these set points. Clear clustering according to cell density was observed by principal component analysis, indicating steady-state dependent metabolite profiles. In particular, varying levels of nucleotides, nucleotide sugar, and lipid precursors explained most of the variance between the different cell density set points. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 2016.

  17. Unusual Stokes V profiles during flaring activity of a delta sunspot

    NASA Astrophysics Data System (ADS)

    Fischer, C. E.; Keller, C. U.; Snik, F.; Fletcher, L.; Socas-Navarro, H.

    2012-11-01

    Aims: We analyze a set of full Stokes profile observations of the flaring active region NOAA 10808. The region was recorded with the Vector-Spectromagnetograph of the Synoptic Optical Long-term Investigations of the Sun facility. The active region produced several successive X-class flares between 19:00 UT and 24:00 UT on September 13, 2005 and we aim to quantify transient and permanent changes in the magnetic field and velocity field during one of the flares, which has been fully captured. Methods: The Stokes profiles were inverted using the height-dependent inversion code LILIA to analyze magnetic field vector changes at the flaring site. We report multilobed asymmetric Stokes V profiles found in the δ-sunspot umbra. We fit the asymmetric Stokes V profiles assuming an atmosphere consisting of two components (SIR inversions) to interpret the profile shape. The results are put in context with Michelson Doppler Imager (MDI) magnetograms and reconstructed X-ray images from the Reuven Ramaty High Energy Solar Spectroscopic Imager. Results: We obtain the magnetic field vector and find signs of restructuring of the photospheric magnetic field during the flare close to the polarity inversion line at the flaring site. At two locations in the umbra we encounter strong fields (~3 kG), as inferred from the Stokes I profiles, which, however, exhibit a low polarization signal. During the flare we observe in addition asymmetric Stokes V profiles at one of these sites. The asymmetric Stokes V profiles appear co-spatial and co-temporal with a strong apparent polarity reversal observed in MDI-magnetograms and a chromospheric hard X-ray source. The two-component atmosphere fits of the asymmetric Stokes profiles result in line-of-sight velocity differences in the range of ~12 km s-1 to 14 km s-1 between the two components in the photosphere. Another possibility is that local atmospheric heating is causing the observed asymmetric Stokes V profile shape. In either case our analysis

  18. Magnetic properties of epitaxial CoCr films with depth-dependent exchange-coupling profiles

    NASA Astrophysics Data System (ADS)

    Fallarino, Lorenzo; Kirby, Brian J.; Pancaldi, Matteo; Riego, Patricia; Balk, Andrew L.; Miller, Casey W.; Vavassori, Paolo; Berger, Andreas

    2017-04-01

    We present a study of the compositional and temperature-dependent magnetic properties of epitaxial CoCr thin films whose composition has a bathtublike depth profile Co /C o1 →1 -xcC r0 →xc/C o1 -xcC rxc/Co1 -x c→1C rxc→0/Co with the highest Cr concentration (xc) at the center of the sample. Polarized neutron reflectometry (PNR) shows that the effective Curie temperature varies as a function of depth and exhibits a minimum in the center of the structure. Correspondingly, we observe that the effective coupling between the two outer Co layers is strongly dependent on the magnetization of the graded CoCr spacer and can be continuously tuned via xc and temperature T . In particular, for xc=0.28 , magnetometry reveals a transition from one-step to two-step reversal behavior for temperatures T > 260 K, indicating a transition from a fully correlated magnetic film structure to an uncoupled system containing effectively two independent magnetic sublayers. Corroborating evidence of the temperature-dependent coupling of the top and bottom regions for xc=0.28 was revealed by PNR, which demonstrated the field-dependent occurrence of antiparallel magnetization alignment on opposite interfaces at sufficiently high temperatures only.

  19. Evaluation of the genetic activity profiles of 65 pesticides

    SciTech Connect

    Garrett, N.E.; Stack, H.F.; Waters, M.D.

    1986-01-01

    This report focuses on the quantitative profiles of genetic activity produced by 65 pesticides and efforts to classify them according to their genotoxic effects and chemical structures. Three main categories may be distinguished based on the qualitative results: Category 1 pesticides were active in most of the in vitro and in vivo assays employed. These nine compounds include the structurally similar organophosphate insecticides, acephate, demeton, monocrotophos, and trichlorfon; the phthalimid fungicide analogues, captan and folpet; and the thiocarbamate herbicide analogues, dialiate, sulfallate and triallate. The 26 Category 2 compounds demonstrated fewer positive results and may be subdivided into two parts, one of which contains 12 halogenated aromatic or heterocyclic ring compounds, including the phenoxy herbicides, 2,4-D, 2,4-DB, and 2,4,5-T. The remaining part of Category 2 (14 compounds) consists of structurally similar organophosphate insecticides, azinphos-methyl, crotoxyphos, disulfoton, methyl parathion; three similar ethylenebisdithiocarbamate fungicides, maneb, mancozeb, and zineb; three similar pyrethroid insecticides, allethrin, chrysanthemic acid, and ethy chrysanthemate; and four structurally diverse compounds, cacodylic acid, dinoseb, sec-butylamine and benomyl. The third category of 30 pesticides gave negative results in all tests and represents structurally diverse compounds. Using the computerized profile matching methodology, from 2080 possible pairwise chemical combinations of the 65 pesticides, 20 statistically significant pairs were selected.

  20. Analysis of communicative behaviour: profiling roles and activities.

    PubMed

    Sørby, Inger Dybdahl; Nytrø, Oystein

    2007-01-01

    In this paper, we discuss how profiles of communicative behaviour can be used to present and analyse information about role activity recorded through structured observation of specific situations. The role activities are encoded as distinctive speech acts. Example profiles resulting from the analysis of three clinicians' communicative behaviour during pre-rounds meetings and ward rounds are given. The examples are based on an observational study performed at a Norwegian university hospital. One fifth-year medical student spent 20 days in two different hospital wards, following 7 physicians from one to seven days each. The observer recorded data from several ward situations such as pre-rounds meetings, ward rounds, and discharge situations. The data was recorded by means of an observation form consisting of a mixture of codes and free-text fields. The data has been post-processed by associating each event with one communicative act. The approach is an efficient and useful means for studying clinicians' information and communication patterns in hospital wards, which can serve as an important tool in the design of new clinical information systems.

  1. Physicochemical signatures of nanoparticle-dependent complement activation

    NASA Astrophysics Data System (ADS)

    Thomas, Dennis G.; Chikkagoudar, Satish; Heredia-Langner, Alejandro; Tardiff, Mark F.; Xu, Zhixiang; Hourcade, Dennis E.; Pham, Christine T. N.; Lanza, Gregory M.; Weinberger, Kilian Q.; Baker, Nathan A.

    2014-01-01

    Nanoparticles are potentially powerful therapeutic tools that have the capacity to target drug payloads and imaging agents. However, some nanoparticles can activate complement, a branch of the innate immune system, and cause adverse side-effects. Recently, we employed an in vitro hemolysis assay to measure the serum complement activity of perfluorocarbon nanoparticles that differed by size, surface charge, and surface chemistry, quantifying the nanoparticle-dependent complement activity using a metric called Residual Hemolytic Activity (RHA). In the present work, we have used a decision tree learning algorithm to derive the rules for estimating nanoparticle-dependent complement response based on the data generated from the hemolytic assay studies. Our results indicate that physicochemical properties of nanoparticles, namely, size, polydispersity index, zeta potential, and mole percentage of the active surface ligand of a nanoparticle, can serve as good descriptors for prediction of nanoparticle-dependent complement activation in the decision tree modeling framework.

  2. Synapse Maturation by Activity-Dependent Ectodomain Shedding of SIRPα

    PubMed Central

    Toth, Anna B.; Terauchi, Akiko; Zhang, Lily Y.; Johnson-Venkatesh, Erin M.; Larsen, David J.; Sutton, Michael A.; Umemori, Hisashi

    2013-01-01

    Formation of appropriate synaptic connections is critical for proper functioning of the brain. After initial synaptic differentiation, active synapses are stabilized by neural activity-dependent signals to establish functional synaptic connections. However, the molecular mechanisms underlying activity-dependent synapse maturation remain to be elucidated. Here we show that activity-dependent ectodomain shedding of SIRPα mediates presynaptic maturation. Two target-derived molecules, FGF22 and SIRPα, sequentially organize the glutamatergic presynaptic terminals during the initial synaptic differentiation and synapse maturation stages, respectively, in the mouse hippocampus. SIRPα drives presynaptic maturation in an activity-dependent fashion. Remarkably, neural activity cleaves the extracellular domain of SIRPα, and the shed ectodomain, in turn, promotes the maturation of the presynaptic terminal. This process involves CaM kinase, matrix metalloproteinases, and the presynaptic receptor CD47. Finally, SIRPα-dependent synapse maturation has significant impacts on synaptic function and plasticity. Thus, ectodomain shedding of SIRPα is an activity-dependent trans-synaptic mechanism for the maturation of functional synapses. PMID:24036914

  3. Improved free energy profile for reduction of NO in cytochrome c dependent nitric oxide reductase (cNOR).

    PubMed

    Blomberg, Margareta R A; Siegbahn, Per E M

    2016-07-15

    Quantum chemical calculations play an essential role in the elucidation of reaction mechanisms for redox-active metalloenzymes. For example, the cleavage and the formation of covalent bonds can usually not be described only on the basis of experimental information, but can be followed by the calculations. Conversely, there are properties, like reduction potentials, which cannot be accurately calculated. Therefore, computational and experimental data has to be carefully combined to obtain reliable descriptions of entire catalytic cycles involving electron and proton uptake from donors outside the enzyme. Such a procedure is illustrated here, for the reduction of nitric oxide (NO) to nitrous oxide and water in the membrane enzyme, cytochrome c dependent nitric oxide reductase (cNOR). A surprising experimental observation is that this reaction is nonelectrogenic, which means that no energy is conserved. On the basis of hybrid density functional calculations a free energy profile for the entire catalytic cycle is obtained, which agrees much better with experimental information on the active site reduction potentials than previous ones. Most importantly the energy profile shows that the reduction steps are endergonic and that the entire process is rate-limited by high proton uptake barriers during the reduction steps. This result implies that, if the reaction were electrogenic, it would become too slow when the gradient is present across the membrane. This explains why this enzyme does not conserve any of the free energy released. © 2016 Wiley Periodicals, Inc.

  4. Linear Energy Transfer-Dependent Change in Rice Gene Expression Profile after Heavy-Ion Beam Irradiation

    PubMed Central

    Ishii, Kotaro; Kazama, Yusuke; Morita, Ryouhei; Hirano, Tomonari; Ikeda, Tokihiro; Usuda, Sachiko; Hayashi, Yoriko; Ohbu, Sumie; Motoyama, Ritsuko; Nagamura, Yoshiaki; Abe, Tomoko

    2016-01-01

    A heavy-ion beam has been recognized as an effective mutagen for plant breeding and applied to the many kinds of crops including rice. In contrast with X-ray or γ-ray, the heavy-ion beam is characterized by a high linear energy transfer (LET). LET is an important factor affecting several aspects of the irradiation effect, e.g. cell survival and mutation frequency, making the heavy-ion beam an effective mutagen. To study the mechanisms behind LET-dependent effects, expression profiling was performed after heavy-ion beam irradiation of imbibed rice seeds. Array-based experiments at three time points (0.5, 1, 2 h after the irradiation) revealed that the number of up- or down-regulated genes was highest 2 h after irradiation. Array-based experiments with four different LETs at 2 h after irradiation identified LET-independent regulated genes that were up/down-regulated regardless of the value of LET; LET–dependent regulated genes, whose expression level increased with the rise of LET value, were also identified. Gene ontology (GO) analysis of LET-independent up-regulated genes showed that some GO terms were commonly enriched, both 2 hours and 3 weeks after irradiation. GO terms enriched in LET-dependent regulated genes implied that some factor regulates genes that have kinase activity or DNA-binding activity in cooperation with the ATM gene. Of the LET-dependent up-regulated genes, OsPARP3 and OsPCNA were identified, which are involved in DNA repair pathways. This indicates that the Ku-independent alternative non-homologous end-joining pathway may contribute to repairing complex DNA legions induced by high-LET irradiation. These findings may clarify various LET-dependent responses in rice. PMID:27462908

  5. Phytochemical profiles and antioxidant activity of adlay varieties.

    PubMed

    Wang, Lifeng; Chen, Jingyi; Xie, Huihui; Ju, Xingrong; Liu, Rui Hai

    2013-05-29

    Consumption of whole grains has been associated with reduced risk of developing major chronic diseases. These health benefits have been attributed in part to their unique phytochemicals. Little is known about the complete profiles of phytochemicals and antioxidant activities of different adlay varieties. The objectives of this study were to determine the phytochemicals profiles of the three adlay varieties, including both free and bound of total phenolics and total flavonoids, and to determine the total antioxidant activity of adlay. The free, bound, and total phenolic contents of adlay samples ranged from 31.23 to 45.19 mg of gallic acid equiv/100 g of sample, from 28.07 to 30.86 mg of gallic acid equiv/100 g of sample, and from 59.30 to 76.04 mg of gallic acid equiv/100 g of sample, respectively. On average, the bound phenolics contributed 45.3% of total phenolic content of the adlay varieties analyzed. The free, bound, and total flavonoid contents of adlay samples ranged from 6.21 to 18.24 mg of catechin equiv/100 g, from 18.68 to 35.27 mg of catechin equiv/100 g, and from 24.88 to 52.86 mg of catechin equiv/100 g, respectively. The average values of bound flavonoids contributed 71.1% of total flavonoids of the adlay varieties analyzed. The percentage contribution of flavonoid content to phenolic content of free, bound, and total ranged from 11.6 to 35.2%, from 50.5 to 66.8%, and from 24.6 to 50.5%. The free, bound, and total oxygen radical absorbance capacity (ORAC) values of adlay samples ranged from 231.9 to 316.6 mg of Trolox equiv/100 g, from 209.0 to 351.4 mg of Trolox equiv/100 g, and from 440.9 to 668.0 mg of Trolox equiv/100 g, respectively. The average ORAC values of bound phytochemicals contributed 48.1% of total antioxidant activity of the adlay varieties analyzed. The content of total polyphenol and the antioxidant capacity are obviously different among different species. Liaoning 5 adlay and Longyi 1 adlay are significantly better than Guizhou

  6. Real-time cell toxicity profiling of Tox21 10K compounds reveals cytotoxicity dependent toxicity pathway linkage.

    PubMed

    Hsieh, Jui-Hua; Huang, Ruili; Lin, Ja-An; Sedykh, Alexander; Zhao, Jinghua; Tice, Raymond R; Paules, Richard S; Xia, Menghang; Auerbach, Scott S

    2017-01-01

    Cytotoxicity is a commonly used in vitro endpoint for evaluating chemical toxicity. In support of the U.S. Tox21 screening program, the cytotoxicity of ~10K chemicals was interrogated at 0, 8, 16, 24, 32, & 40 hours of exposure in a concentration dependent fashion in two cell lines (HEK293, HepG2) using two multiplexed, real-time assay technologies. One technology measures the metabolic activity of cells (i.e., cell viability, glo) while the other evaluates cell membrane integrity (i.e., cell death, flor). Using glo technology, more actives and greater temporal variations were seen in HEK293 cells, while results for the flor technology were more similar across the two cell types. Chemicals were grouped into classes based on their cytotoxicity kinetics profiles and these classes were evaluated for their associations with activity in the Tox21 nuclear receptor and stress response pathway assays. Some pathways, such as the activation of H2AX, were associated with the fast-responding cytotoxicity classes, while others, such as activation of TP53, were associated with the slow-responding cytotoxicity classes. By clustering pathways based on their degree of association to the different cytotoxicity kinetics labels, we identified clusters of pathways where active chemicals presented similar kinetics of cytotoxicity. Such linkages could be due to shared underlying biological processes between pathways, for example, activation of H2AX and heat shock factor. Others involving nuclear receptor activity are likely due to shared chemical structures rather than pathway level interactions. Based on the linkage between androgen receptor antagonism and Nrf2 activity, we surmise that a subclass of androgen receptor antagonists cause cytotoxicity via oxidative stress that is associated with Nrf2 activation. In summary, the real-time cytotoxicity screen provides informative chemical cytotoxicity kinetics data related to their cytotoxicity mechanisms, and with our analysis, it is

  7. Real-time cell toxicity profiling of Tox21 10K compounds reveals cytotoxicity dependent toxicity pathway linkage

    PubMed Central

    Huang, Ruili; Lin, Ja-An; Sedykh, Alexander; Zhao, Jinghua; Tice, Raymond R.; Paules, Richard S.; Xia, Menghang; Auerbach, Scott S.

    2017-01-01

    Cytotoxicity is a commonly used in vitro endpoint for evaluating chemical toxicity. In support of the U.S. Tox21 screening program, the cytotoxicity of ~10K chemicals was interrogated at 0, 8, 16, 24, 32, & 40 hours of exposure in a concentration dependent fashion in two cell lines (HEK293, HepG2) using two multiplexed, real-time assay technologies. One technology measures the metabolic activity of cells (i.e., cell viability, glo) while the other evaluates cell membrane integrity (i.e., cell death, flor). Using glo technology, more actives and greater temporal variations were seen in HEK293 cells, while results for the flor technology were more similar across the two cell types. Chemicals were grouped into classes based on their cytotoxicity kinetics profiles and these classes were evaluated for their associations with activity in the Tox21 nuclear receptor and stress response pathway assays. Some pathways, such as the activation of H2AX, were associated with the fast-responding cytotoxicity classes, while others, such as activation of TP53, were associated with the slow-responding cytotoxicity classes. By clustering pathways based on their degree of association to the different cytotoxicity kinetics labels, we identified clusters of pathways where active chemicals presented similar kinetics of cytotoxicity. Such linkages could be due to shared underlying biological processes between pathways, for example, activation of H2AX and heat shock factor. Others involving nuclear receptor activity are likely due to shared chemical structures rather than pathway level interactions. Based on the linkage between androgen receptor antagonism and Nrf2 activity, we surmise that a subclass of androgen receptor antagonists cause cytotoxicity via oxidative stress that is associated with Nrf2 activation. In summary, the real-time cytotoxicity screen provides informative chemical cytotoxicity kinetics data related to their cytotoxicity mechanisms, and with our analysis, it is

  8. Activity-dependent inhibitory synapse remodeling through gephyrin phosphorylation.

    PubMed

    Flores, Carmen E; Nikonenko, Irina; Mendez, Pablo; Fritschy, Jean-Marc; Tyagarajan, Shiva K; Muller, Dominique

    2015-01-06

    Maintaining a proper balance between excitation and inhibition is essential for the functioning of neuronal networks. However, little is known about the mechanisms through which excitatory activity can affect inhibitory synapse plasticity. Here we used tagged gephyrin, one of the main scaffolding proteins of the postsynaptic density at GABAergic synapses, to monitor the activity-dependent adaptation of perisomatic inhibitory synapses over prolonged periods of time in hippocampal slice cultures. We find that learning-related activity patterns known to induce N-methyl-D-aspartate (NMDA) receptor-dependent long-term potentiation and transient optogenetic activation of single neurons induce within hours a robust increase in the formation and size of gephyrin-tagged clusters at inhibitory synapses identified by correlated confocal electron microscopy. This inhibitory morphological plasticity was associated with an increase in spontaneous inhibitory activity but did not require activation of GABAA receptors. Importantly, this activity-dependent inhibitory plasticity was prevented by pharmacological blockade of Ca(2+)/calmodulin-dependent protein kinase II (CaMKII), it was associated with an increased phosphorylation of gephyrin on a site targeted by CaMKII, and could be prevented or mimicked by gephyrin phospho-mutants for this site. These results reveal a homeostatic mechanism through which activity regulates the dynamics and function of perisomatic inhibitory synapses, and they identify a CaMKII-dependent phosphorylation site on gephyrin as critically important for this process.

  9. Angular dependence of Doppler profiles of atomic emission produced in electron-molecule collisions: Estimation of anisotropy parameters

    NASA Astrophysics Data System (ADS)

    Nakashima, Keiji; Ogawa, Teiichiro

    1985-11-01

    The angular dependence of Doppler profiles of atomic fluorescence produced in electron impact dissociation of molecules was simulated in consideration of the effect of the anisotropy of dissociation and the ``polarization'' in magnetic sublevel. The asymmetry parameter b and the polarization of the electric vector of emission Jp are key parameters of Doppler profiles for the excited atom of known translational energy distribution. The difference of two Doppler profiles taken at 90° and 45°, which is denoted as angular difference Doppler profile, is shown to be useful to estimate these two key parameters.

  10. Molecular Profiling of Activated Neurons by Phosphorylated Ribosome Capture

    PubMed Central

    Knight, Zachary A.; Tan, Keith; Birsoy, Kivanc; Schmidt, Sarah; Garrison, Jennifer L.; Wysocki, Robert W.; Emiliano, Ana; Ekstrand, Mats I.; Friedman, Jeffrey M.

    2013-01-01

    SUMMARY The mammalian brain is composed of thousands of interacting neural cell types. Systematic approaches to establish the molecular identity of functional populations of neurons would advance our understanding of neural mechanisms controlling behavior. Here, we show that ribosomal protein S6, a structural component of the ribosome, becomes phosphorylated in neurons activated by a wide range of stimuli. We show that these phosphorylated ribosomes can be captured from mouse brain homogenates, thereby enriching directly for the mRNAs expressed in discrete subpopulations of activated cells. We use this approach to identify neurons in the hypothalamus regulated by changes in salt balance or food availability. We show that galanin neurons are activated by fasting and that prodynorphin neurons restrain food intake during scheduled feeding. These studies identify elements of the neural circuit that controls food intake and illustrate how the activity-dependent capture of cell-type-specific transcripts can elucidate the functional organization of a complex tissue. PMID:23178128

  11. Metabolomic profiling and antioxidant activity of some Acacia species

    PubMed Central

    Abdel-Farid, I.B.; Sheded, M.G.; Mohamed, E.A.

    2014-01-01

    Metabolomic profiling of different parts (leaves, flowers and pods) of Acacia species (Acacia nilotica, Acacia seyal and Acacia laeta) was evaluated. The multivariate data analyses such as principal component analysis (PCA) and partial least square-discriminant analysis (PLS-DA) were used to differentiate the distribution of plant metabolites among different species or different organs of the same species. A.nilotica was characterized with a high content of saponins and A.seyal was characterized with high contents of proteins, phenolics, flavonoids and anthocyanins. A.laeta had a higher content of carbohydrates than A. nilotica and A. seyal. On the basis of these results, total antioxidant capacity, DPPH free radical scavenging activity and reducing power of the methanolic extracts of studied parts were evaluated. A.nilotica and A.seyal extracts showed less inhibitory concentration 50 (IC50) compared to A.laeta extracts which means that these two species have the strongest radical scavenging activity whereas A. laeta extracts have the lowest radical scavenging activity. A positive correlation between saponins and flavonoids with total antioxidant capacity and DPPH radical scavenging activity was observed. Based on these results, the potentiality of these plants as antioxidants was discussed. PMID:25313274

  12. Activity-based probes for the proteomic profiling of metalloproteases

    PubMed Central

    Saghatelian, Alan; Jessani, Nadim; Joseph, Arul; Humphrey, Mark; Cravatt, Benjamin F.

    2004-01-01

    Metalloproteases (MPs) are a large and diverse class of enzymes implicated in numerous physiological and pathological processes, including tissue remodeling, peptide hormone processing, and cancer. MPs are tightly regulated by multiple posttranslational mechanisms in vivo, hindering their functional analysis by conventional genomic and proteomic methods. Here we describe a general strategy for creating activity-based proteomic probes for MPs by coupling a zinc-chelating hydroxamate to a benzophenone photocrosslinker, which promote selective binding and modification of MP active sites, respectively. These probes labeled active MPs but not their zymogen or inhibitor-bound counterparts and were used to identify members of this enzyme class up-regulated in invasive cancer cells and to evaluate the selectivity of MP inhibitors in whole proteomes. Interestingly, the matrix metalloproteinase inhibitor GM6001 (ilomastat), which is currently in clinical development, was found to also target the neprilysin, aminopeptidase, and dipeptidylpeptidase clans of MPs. These results demonstrate that MPs can display overlapping inhibitor sensitivities despite lacking sequence homology and stress the need to evaluate MP inhibitors broadly across this enzyme class to develop agents with suitable target selectivities in vivo. Activity-based profiling offers a powerful means for conducting such screens, as this approach can be carried out directly in whole proteomes, thereby facilitating the discovery of disease-associated MPs concurrently with inhibitors that selectively target these proteins. PMID:15220480

  13. Occipital TMS has an activity-dependent suppressive effect

    PubMed Central

    Perini, Francesca; Cattaneo, Luigi; Carrasco, Marisa; Schwarzbach, Jens V.

    2012-01-01

    The effects of transcranial magnetic stimulation (TMS) vary depending on the brain state at the stimulation moment. Four mechanisms have been proposed to underlie these effects: (i) virtual lesion–TMS suppresses neural signals; (ii) preferential activation of less active neurons–TMS drives up activity in the stimulated area, but active neurons are saturating, (iii) noise generation–TMS adds random neuronal activity and its effect interacts with stimulus-intensity; (iv) noise generation–TMS adds random neuronal activity and its effect depends on TMS-intensity. Here we explore these hypotheses by investigating the effects of TMS on early visual cortex on the contrast response function while varying adaptation state of the observers. We tested human participants in an orientation discrimination task, in which performance is contingent upon contrast sensitivity. Before each trial, neuronal activation of visual cortex was altered through contrast adaptation to two flickering gratings. In a factorial design, with or without adaptation, a single TMS pulse was delivered simultaneously with targets of varying contrast. Adaptation decreased contrast sensitivity. The effect of TMS on performance was state-dependent: TMS decreased contrast sensitivity in the absence of adaptation but increased it after adaptation. None of the proposed mechanisms can account for the results in their entirety, in particular, for the facilitatory effect at intermediate to high contrasts after adaptation. We propose an alternative hypothesis: TMS effects are activity-dependent, so that TMS suppresses the most active neurons and thereby changes the balance between excitation and inhibition. PMID:22956826

  14. Muscle Transcriptional Profile Based on Muscle Fiber, Mitochondrial Respiratory Activity, and Metabolic Enzymes

    PubMed Central

    Liu, Xuan; Du, Yang; Trakooljul, Nares; Brand, Bodo; Muráni, Eduard; Krischek, Carsten; Wicke, Michael; Schwerin, Manfred; Wimmers, Klaus; Ponsuksili, Siriluck

    2015-01-01

    Skeletal muscle is a highly metabolically active tissue that both stores and consumes energy. Important biological pathways that affect energy metabolism and metabolic fiber type in muscle cells may be identified through transcriptomic profiling of the muscle, especially ante mortem. Here, gene expression was investigated in malignant hyperthermia syndrome (MHS)-negative Duroc and Pietrian (PiNN) pigs significantly differing for the muscle fiber types slow-twitch-oxidative fiber (STO) and fast-twitch-oxidative fiber (FTO) as well as mitochondrial activity (succinate-dependent state 3 respiration rate). Longissimus muscle samples were obtained 24 h before slaughter and profiled using cDNA microarrays. Differential gene expression between Duroc and PiNN muscle samples were associated with protein ubiquitination, stem cell pluripotency, amyloid processing, and 3-phosphoinositide biosynthesis and degradation pathways. In addition, weighted gene co-expression network analysis within both breeds identified several co-expression modules that were associated with the proportion of different fiber types, mitochondrial respiratory activity, and ATP metabolism. In particular, Duroc results revealed strong correlations between mitochondrion-associated co-expression modules and STO (r = 0.78), fast-twitch glycolytic fiber (r = -0.98), complex I (r=0.72) and COX activity (r = 0.86). Other pathways in the protein-kinase-activity enriched module were positively correlated with STO (r=0.93), while negatively correlated with FTO (r = -0.72). In contrast to PiNN, co-expression modules enriched in macromolecule catabolic process, actin cytoskeleton, and transcription activator activity were associated with fiber types, mitochondrial respiratory activity, and metabolic enzyme activities. Our results highlight the importance of mitochondria for the oxidative capacity of porcine muscle and for breed-dependent molecular pathways in muscle cell fibers. PMID:26681915

  15. Active flow control for a NACA-0012 Profile: Part II

    NASA Astrophysics Data System (ADS)

    Oualli, H.; Makadem, M.; Ouchene, H.; Ferfouri, A.; Bouabdallah, A.; Gad-El-Hak, M.

    2016-11-01

    Active flow control is applied to a NACA-0012 profile. The experiments are conducted in a wind tunnel. Using a high-resolution visible-light camera and tomography, flow visualizations are carried out. LES finite-volume 3D code is used to complement the physical experiments. The symmetric wing is clipped into two parts, and those parts extend and retract along the chord according to the same sinusoidal law we optimized last year for the same profile but clipped at an angle of 60 deg, instead of the original 90 deg. The Reynolds number range is extended to 500,000, thus covering the flying regimes of micro-UAVs, UAVs, as well as small aircraft. When the nascent cavity is open and the attack angle is 30 deg, the drag coefficient is increased by 1,300%, as compared to the uncontrolled case. However, when the cavity is covered and Re <=105 , a relatively small frequency, f <= 30 Hz, is required for the drag coefficient to drop to negative values. At the maximum Reynolds number, thrust is generated but only at much higher frequencies, 12 <= f <= 16 kHz.

  16. Solar activity effects on the equatorial thermosphere temperature profile

    NASA Astrophysics Data System (ADS)

    Arduini, C.; Laneve, G.; Nobile, L.

    In this paper we present the effects of solar activity on the temperature profiles of the equatorial thermosphere as derived from the neutral density data collected by the San Marco 5 (SM5) satellite. This satellite flew during the increasing part of the solar cycle 22 (1988). It had a quasi-equatorial orbit, with inclination lower than 3 deg. The range of measurements, from April to December, allows the inference of seasonal and diurnal effects on the temperature profiles. The density data are collected every second along arcs of orbit lasting up to 50 minutes. The analysis of these densities has been already partially presented and provided evidence for several interesting features, in particular the vertical structure of the diurnal harmonic content and its seasonal variations. The temperatures derived from the same data set provide a useful complement to this picture. The SM5 satellite carried on board 5 instruments for studying the equatorial ionosphere and thermosphere, among them, the Drag Balance Instrument (DBI) for measuring the neutral density and the Ion Drift Meter and Potential Retarding Analyzer (IVI) that allow the evaluation of ions concentration, velocity and temperature. It is possible, therefore, to compare the neutral temperature derived from the neutral density data with the ion temperature given by the IVI.

  17. Subgroup-dependent effects of voluntary alcohol intake on behavioral profiles in outbred Wistar rats.

    PubMed

    Momeni, Shima; Roman, Erika

    2014-12-15

    Experimental animal models are critical for understanding the genetic, environmental and neurobiological underpinnings of alcohol use disorders. Limited studies investigate alcohol-induced effects on behavior using free-choice paradigms. The aims of the present experiment were to study voluntary alcohol intake using a modified intermittent access paradigm, investigate the effects of voluntary alcohol intake on behavioral profiles in water- and alcohol-drinking rats, and select extreme low- and high-drinking animals for a more detailed behavioral characterization. Sixty outbred male Wistar rats were randomized into water and alcohol groups. Behavioral profiles in the multivariate concentric square field™ (MCSF) test were assessed prior to and after voluntary alcohol intake. The animals had intermittent access to 20% alcohol and water for three consecutive days per week for seven weeks. The results revealed increased alcohol intake over time. No major alcohol-induced differences on behavior profiles were found when comparing water- and alcohol-drinking animals. The high-drinking animals displayed an alcohol deprivation effect, which was not found in the low-drinking animals. High-drinking rats had lower risk-taking behavior prior to alcohol access and lower anxiety-like behavior after voluntary alcohol intake compared to low-drinking rats. In conclusion, the modified intermittent access paradigm may be useful for pharmacological manipulation of alcohol intake. With regard to behavior, the present findings highlights the importance of studying subgroup-dependent differences and add to the complexity of individual differences in behavioral traits of relevance to the vulnerability for excessive alcohol intake.

  18. Wavelength dependent near-range lidar profiling of smog aerosol over Athens

    NASA Astrophysics Data System (ADS)

    Stachlewska, Iwona S.; Marinou, Eleni; Engelmann, Ronny; Costa Surós, Montserrat; Kottas, Mickael; Baars, Holger; Janicka, Lucja; Solomos, Stavros; Heese, Birgit; Kumala, Wojciech; Tsekeri, Alexandra; Binietoglou, Ioannis; Markowicz, Krzysztof M.; Amiridis, Vassilis; Balis, Dimitris; Althausen, Dietrich; Wandinger, Ulla; Ansmann, Albert

    2016-04-01

    Recently, the ACTRIS2 JRA1 field campaign focusing on joint remote and in-situ sensing of absorbing aerosols has been conducted in Athens (http://actris-athens.eu). In the frame of the ACTRIS2 BL-Smog TNA, co-located measurements of the near-range lidar receiver (NARLa) of the University of Warsaw with the multi-wavelength PollyXT lidar of the National Observatory of Athens were performed. The excellent capacities of the PollyXT-NOA lidar, equipped with eight far-range channels (355, 355s, 387, 407, 532, 532s, 607, and 1064nm) and two near-range channels (532 and 607 nm), were enhanced by integrating the NARLa for simultaneous observations. By using the NARLa, equipped with the elastic channels (355 and 532nm) and Raman channels (387 and 607nm), the wavelength dependence of the aerosol particles properties within boundary layer was captured. The dominant conditions observed during the JRA1 period were the fresh winter smog layers occurring in lowermost boundary layer over Athens. NARLa provided profiles as close to surface as 50m, thus the data obtained in the near-range were used for the incomplete overlap region of the far-field channels. With NARLa we assessed the overlap at 355 and 532nm wavelengths and concluded on the possibility of using the single near-range 532 nm channel for the overlap correction in both VIS and UV channels of the PollyXT-NOA. As a result, the obtained lidar profiles are expected to be more consistent with the sunphotometer measurements. In the future, the GARRLiC code can be applied on the synergy of combined near and far range lidar profiles with AERONET data sets in order to study improvement on the inversion results.

  19. Metabolic profiling in Caenorhabditis elegans provides an unbiased approach to investigations of dosage dependent lead toxicity.

    PubMed

    Sudama, Gita; Zhang, John; Isbister, Jenefir; Willett, James D

    2013-02-01

    The nematode, Caenorhabditis elegans (CE), serves as a model system in which to explore the impact of particularly low-levels of lead [250, 500, 1000 and 2000 parts per million (ppm) (1.4 × 10(-6) M to 1.1 × 10(-5) M/nematode)] on specific metabolic pathways and processes. Chromatographic profiles of redox active metabolites are captured through application of high performance liquid chromatography coupled to electrochemical detection (Coularray/HPLC). Principal Component Analysis (PCA: unbiased cluster analysis) and the application of a slicing program, located significant areas of difference occurring within the 2.8-4.58 min section of the chromatograms. It is within this region of the data profiles that known components of the purine pathway reside. Two analytes of unknown structure were detected at 3.5 and 4 min respectively. Alterations in levels of the purine, tryptophan and tyrosine pathway intermediates measured in response to differing concentrations of lead acetate indicate that the effect of lead on these pathways is not linear, yet the ratio of the pathway precursors, tryptophan and tyrosine remains relatively constant. The application of the above combined analytical approaches enhances the value of data generated. Exposure of CE to very low levels of lead produced significant alterations in profiles of electrochemically active compounds. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s11306-012-0438-0) contains supplementary material, which is available to authorized users.

  20. Diagnostic and therapeutic approaches in respiratory allergy are different depending on the profile of aeroallergen sensitisation.

    PubMed

    Domínguez-Ortega, J; Quirce, S; Delgado, J; Dávila, I; Martí-Guadaño, E; Valero, A

    2014-01-01

    There are few studies which analyse the characteristics of allergic respiratory disease according to profiles of sensitisation to different allergens. This study describes the clinical features and therapeutic approaches, according to the sensitisation profile to relevant aeroallergens, in a sample of adult patients with a first-time diagnosis of respiratory allergy (rhinitis and/or asthma). 1287 patients, enrolled consecutively in the spring of 2010 by 200 allergy specialists, were classified into four groups according to sensitisation to significant allergens in each geographical area (grass pollen, olive pollen, grass and olive pollen, house dust mites). Information was obtained on demographics, diagnostic procedures used, treatments prescribed, clinical characteristics of the rhinitis, and severity and control of asthma. Of the patients, 58.6% had rhinitis only and 38.7% had both rhinitis and asthma. Patients with more severe rhinitis had more severe and poorer controlled asthma. Sensitisation to different allergens was not associated with significant differences in severity and control of asthma, but patients with house dust mite allergy presented persistent rhinitis more frequently. Allergy to grass pollen was significantly associated with food allergies. Differences were observed in the frequency of prescription of immunotherapy and antileukotrienes in patients allergic to house dust mites and of topical corticosteroids in patients with pollen allergy. It was observed in this study that in respiratory allergy disease, there are clinical differences as well as differences in diagnostic procedure and therapeutic attitudes, depending on the clinically relevant allergen. Copyright © 2012 SEICAP. Published by Elsevier Espana. All rights reserved.

  1. Profile of deferasirox for the treatment of patients with non-transfusion-dependent thalassemia syndromes

    PubMed Central

    Ricchi, Paolo; Marsella, Maria

    2015-01-01

    It has been clearly shown that iron overload adds progressively significant morbidity and mortality in patients with non-transfusion-dependent thalassemia (NTDT). The lack of physiological mechanisms to eliminate the excess of iron requires effective iron chelation therapy. The reduced compliance to deferoxamine and the risk of severe hematological adverse events during deferiprone treatment have limited the use of both these drugs to correct iron imbalance in NTDT. According to the principles of evidence-based medicine, following the demonstration of the effectiveness and the safety of deferasirox (Exjade®) in a prospective, randomized, controlled trial, deferasirox was approved by the US Food and Drug Administration in May 2013 for the treatment of iron overload associated with NTDT. This review, assessing the available scientific literature, will focus on the profile of DFX in the treatment of non-transfusional hemosiderosis in patients with NTDT. PMID:26719673

  2. Metabolomic Profiling of Hormone-Dependent Cancers: A Bird’s Eye View

    PubMed Central

    Lloyd, Stacy M.; Arnold, James; Sreekumar, Arun

    2015-01-01

    Hormone-dependent cancers present a significant public health challenge, as they are among the most common cancers in the world. One factor associated with cancer development and progression is metabolic reprogramming. By understanding these alterations, we can identify potential markers and novel biochemical therapeutic targets. Metabolic profiling is an advanced technology that allows investigators to assess low molecular weight compounds that reflect physiological alterations. Current research in metabolomics in prostate and breast cancer has made great strides in uncovering specific metabolic pathways that are associated with cancer development, progression, and resistance. This review will highlight some of the major findings and potential therapeutic advances that have been reported utilizing this technology. PMID:26242817

  3. Iterative approach to self-adapting and altitude-dependent regularization for atmospheric profile retrievals.

    PubMed

    Ridolfi, Marco; Sgheri, Luca

    2011-12-19

    In this paper we present the IVS (Iterative Variable Strength) method, an altitude-dependent, self-adapting Tikhonov regularization scheme for atmospheric profile retrievals. The method is based on a similar scheme we proposed in 2009. The new method does not need any specifically tuned minimization routine, hence it is more robust and faster. We test the self-consistency of the method using simulated observations of the Michelson Interferometer for Passive Atmospheric Sounding (MIPAS). We then compare the new method with both our previous scheme and the scalar method currently implemented in the MIPAS on-line processor, using both synthetic and real atmospheric limb measurements. The IVS method shows very good performances.

  4. Profile of deferasirox for the treatment of patients with non-transfusion-dependent thalassemia syndromes.

    PubMed

    Ricchi, Paolo; Marsella, Maria

    2015-01-01

    It has been clearly shown that iron overload adds progressively significant morbidity and mortality in patients with non-transfusion-dependent thalassemia (NTDT). The lack of physiological mechanisms to eliminate the excess of iron requires effective iron chelation therapy. The reduced compliance to deferoxamine and the risk of severe hematological adverse events during deferiprone treatment have limited the use of both these drugs to correct iron imbalance in NTDT. According to the principles of evidence-based medicine, following the demonstration of the effectiveness and the safety of deferasirox (Exjade(®)) in a prospective, randomized, controlled trial, deferasirox was approved by the US Food and Drug Administration in May 2013 for the treatment of iron overload associated with NTDT. This review, assessing the available scientific literature, will focus on the profile of DFX in the treatment of non-transfusional hemosiderosis in patients with NTDT.

  5. Thickness dependent CARS measurement of polymeric thin films without depth-profiling.

    PubMed

    Choi, Dae Sik; Jeoung, Sae Chae; Chon, Byung-Hyuk

    2008-02-18

    Coherent anti-Stokes Raman scattering (CARS) microscopy is demonstrated to be a promising optical method for the characterization of polymer films with film thickness varying between 180 nm to 4300 nm. In case of PMMA films with a thickness of few hundreds of nanometers, the observed CARS signal was mainly associated with the interference effect of large nonresonant CARS field from glass substrate and the weak resonant field of PMMA. The dependence of resonant CARS intensity of PMMA film on film thickness is in good agreement with the theoretical prediction on a CARS field. The current work offers potential possibilities of noninvasive thickness measurement of polymeric thin film of thickness less than 180 nm by multiplex CARS microscopy without depth-profiling.

  6. Comparison of phytochemical profiles, antioxidant and cellular antioxidant activities of different varieties of blueberry (Vaccinium spp.).

    PubMed

    Wang, Huailing; Guo, Xinbo; Hu, Xiaodan; Li, Tong; Fu, Xiong; Liu, Rui Hai

    2017-02-15

    Numerous reports have demonstrated that the consumption of fruits and vegetables is beneficial for the human health. Blueberries, in particular, are rich in phytochemicals including free and bound forming. Phytochemical profiles of 14 varieties of blueberry were compared in this study. 12 compounds were analyzed and had significant changes in blueberry fruits. Total antioxidant activities in different blueberry varieties varied about 2.6times by oxygen radical absorbance capacity (ORAC) assay, and 2times by peroxyl radical scavenging capacity (PSC) assay. The cellular antioxidant activities (CAA) in different varieties varied about 3.9times without phosphate buffer saline (PBS) wash, and 4.7times with PBS wash by CAA assay. Blueberry extracts had potent antiproliferative activities against HepG2 human liver cancer cells, indicating the potential protective benefits associated with their use as functional foods. The anti-proliferative activity was observed to be dose-dependent in blueberry extracts.

  7. Efficacy of treatment in an opioid -dependent population group using the Maudsley Addiction Profile (MAP) tool.

    PubMed

    Collins, Ruth; Boggs, Bob; Taggart, Noel; Kelly, Martin; Drillington, Aileen; Swanton, Ivy; Patterson, Diane

    2009-01-01

    A pilot study was performed to assess the effectiveness of treatment in an opioid dependent population using the Maudsley Addiction Profile (MAP) tool1.The primary outcome of the study was to assess if treatment had an effect on 1. Substance use (quantity and frequency of use), 2. Health risk behaviour (injecting and sharing injecting equipment), 3. Health symptoms (physical and psychological) and 4. Personal /Social functioning (relationships, employment and crime). A secondary outcome was also sought.The study took place in 2007 in an inner city Belfast hospital specialising in the treatment of addiction, over a two month period. Fifteen patients, all opioid dependent and receiving outpatient community treatment, were interviewed at baseline (prior to the commencement of treatment) and at eight weeks follow up.Three patients were lost to follow up. Two patients stopped using altogether. Of the remaining patients, improvements were seen in most areas. There was a decrease in the use of heroin (71.28%), cocaine (99.72%), crack cocaine (100%), cannabis (99.94%) and alcohol (33.17%). There was a reduction in injecting behaviour (60.93%). Improvements were observed in health with a reduction in physical (41.35%) and psychological (35%) symptoms. Overall personal and social functioning improved regarding interactions with family and friends. A reduction in crime was also observed (75%).Opinions and views of staff involved in the study were generally positive.This patient population presents with multiple and complex needs. Effective treatment needs to address these needs and not just drug addiction alone. The Maudsley Addiction Profile tool highlights this.

  8. Lexical learning in mild aphasia: gesture benefit depends on patholinguistic profile and lesion pattern.

    PubMed

    Kroenke, Klaus-Martin; Kraft, Indra; Regenbrecht, Frank; Obrig, Hellmuth

    2013-01-01

    Gestures accompany speech and enrich human communication. When aphasia interferes with verbal abilities, gestures become even more relevant, compensating for and/or facilitating verbal communication. However, small-scale clinical studies yielded diverging results with regard to a therapeutic gesture benefit for lexical retrieval. Based on recent functional neuroimaging results, delineating a speech-gesture integration network for lexical learning in healthy adults, we hypothesized that the commonly observed variability may stem from differential patholinguistic profiles in turn depending on lesion pattern. Therefore we used a controlled novel word learning paradigm to probe the impact of gestures on lexical learning, in the lesioned language network. Fourteen patients with chronic left hemispheric lesions and mild residual aphasia learned 30 novel words for manipulable objects over four days. Half of the words were trained with gestures while the other half were trained purely verbally. For the gesture condition, rootwords were visually presented (e.g., Klavier, [piano]), followed by videos of the corresponding gestures and the auditory presentation of the novel words (e.g., /krulo/). Participants had to repeat pseudowords and simultaneously reproduce gestures. In the verbal condition no gesture-video was shown and participants only repeated pseudowords orally. Correlational analyses confirmed that gesture benefit depends on the patholinguistic profile: lesser lexico-semantic impairment correlated with better gesture-enhanced learning. Conversely largely preserved segmental-phonological capabilities correlated with better purely verbal learning. Moreover, structural MRI-analysis disclosed differential lesion patterns, most interestingly suggesting that integrity of the left anterior temporal pole predicted gesture benefit. Thus largely preserved semantic capabilities and relative integrity of a semantic integration network are prerequisites for successful use of

  9. Comparative transcriptome profiling of the injured zebrafish and mouse hearts identifies miRNA-dependent repair pathways

    PubMed Central

    Crippa, Stefania; Nemir, Mohamed; Ounzain, Samir; Ibberson, Mark; Berthonneche, Corinne; Sarre, Alexandre; Boisset, Gaëlle; Maison, Damien; Harshman, Keith; Xenarios, Ioannis; Diviani, Dario; Schorderet, Daniel; Pedrazzini, Thierry

    2016-01-01

    Aims The adult mammalian heart has poor regenerative capacity. In contrast, the zebrafish heart retains a robust capacity for regeneration into adulthood. These distinct responses are consequences of a differential utilization of evolutionary-conserved gene regulatory networks in the damaged heart. To systematically identify miRNA-dependent networks controlling cardiac repair following injury, we performed comparative gene and miRNA profiling of the cardiac transcriptome in adult mice and zebrafish. Methods and results Using an integrated approach, we show that 45 miRNA-dependent networks, involved in critical biological pathways, are differentially modulated in the injured zebrafish vs. mouse hearts. We study, more particularly, the miR-26a-dependent response. Therefore, miR-26a is down-regulated in the fish heart after injury, whereas its expression remains constant in the mouse heart. Targets of miR-26a involve activators of the cell cycle and Ezh2, a component of the polycomb repressive complex 2 (PRC2). Importantly, PRC2 exerts repressive functions on negative regulators of the cell cycle. In cultured neonatal cardiomyocytes, inhibition of miR-26a stimulates, therefore, cardiomyocyte proliferation. Accordingly, miR-26a knockdown prolongs the proliferative window of cardiomyocytes in the post-natal mouse heart. Conclusions This novel strategy identifies a series of miRNAs and associated pathways, in particular miR-26a, which represent attractive therapeutic targets for inducing repair in the injured heart. PMID:26857418

  10. RNA-Dependent RNA Polymerase Activity in Influenza Virions

    PubMed Central

    Penhoet, Edward; Miller, Henry; Doyle, Michael; Blatti, Stanley

    1971-01-01

    An RNA-dependent RNA polymerase activity has been detected in purified preparations of influenza virus. In contrast to the replicase activity induced in influenza-infected cells, the virion-associated enzyme has an absolute requirement for Mn++. Most of the RNA synthesized in vitro is complementary to virion RNA. PMID:5288388

  11. Influence of Saline on Temperature Profile of Laser Lithotripsy Activation

    PubMed Central

    Silva, Igor N.; Donalisio da Silva, Rodrigo; Gustafson, Diedra; Sehrt, David; Kim, Fernando J.

    2015-01-01

    Abstract Purpose: We established an ex vivo model to evaluate the temperature profile of the ureter during laser lithotripsy, the influence of irrigation on temperature, and thermal spread during lithotripsy with the holmium:yttrium-aluminum-garnet (Ho:YAG) laser. Materials and Methods: Two ex vivo models of Ovis aries urinary tract and human calcium oxalate calculi were used. The Open Ureteral Model was opened longitudinally to measure the thermal profile of the urothelium. On the Clinical Model, anterograde ureteroscopy was performed in an intact urinary system. Temperatures were measured on the external portion of the ureter and the urothelium during lithotripsy and intentional perforation. The lithotripsy group (n=20) was divided into irrigated (n=10) and nonirrigated (n=10), which were compared for thermal spread length and values during laser activation. The intentional perforation group (n=10) was evaluated under saline flow. The Ho:YAG laser with a 365 μm laser fiber and power at 10W was used (1J/Pulse at 10 Hz). Infrared Fluke Ti55 Thermal Imager was used for evaluation. Maximum temperature values were recorded and compared. Results: On the Clinical Model, the external ureteral wall obtained a temperature of 37.4°C±2.5° and 49.5°C±2.3° (P=0.003) and in the Open Ureteral Model, 49.7°C and 112.4°C with and without irrigation, respectively (P<0.05). The thermal spread along the external ureter wall was not statically significant with or without irrigation (P=0.065). During intentional perforation, differences in temperatures were found between groups (opened with and without irrigation): 81.8°±8.8° and 145.0°±15.0°, respectively (P<0.005). Conclusion: There is an increase in the external ureteral temperature during laser activation, but ureteral thermal values decreased when saline flow was applied. Ureter thermal spread showed no difference between irrigated and nonirrigated subgroups. This is the first laser lithotripsy thermography study

  12. Influence of saline on temperature profile of laser lithotripsy activation.

    PubMed

    Molina, Wilson R; Silva, Igor N; Donalisio da Silva, Rodrigo; Gustafson, Diedra; Sehrt, David; Kim, Fernando J

    2015-02-01

    We established an ex vivo model to evaluate the temperature profile of the ureter during laser lithotripsy, the influence of irrigation on temperature, and thermal spread during lithotripsy with the holmium:yttrium-aluminum-garnet (Ho:YAG) laser. Two ex vivo models of Ovis aries urinary tract and human calcium oxalate calculi were used. The Open Ureteral Model was opened longitudinally to measure the thermal profile of the urothelium. On the Clinical Model, anterograde ureteroscopy was performed in an intact urinary system. Temperatures were measured on the external portion of the ureter and the urothelium during lithotripsy and intentional perforation. The lithotripsy group (n=20) was divided into irrigated (n=10) and nonirrigated (n=10), which were compared for thermal spread length and values during laser activation. The intentional perforation group (n=10) was evaluated under saline flow. The Ho:YAG laser with a 365 μm laser fiber and power at 10W was used (1J/Pulse at 10 Hz). Infrared Fluke Ti55 Thermal Imager was used for evaluation. Maximum temperature values were recorded and compared. On the Clinical Model, the external ureteral wall obtained a temperature of 37.4°C±2.5° and 49.5°C±2.3° (P=0.003) and in the Open Ureteral Model, 49.7°C and 112.4°C with and without irrigation, respectively (P<0.05). The thermal spread along the external ureter wall was not statically significant with or without irrigation (P=0.065). During intentional perforation, differences in temperatures were found between groups (opened with and without irrigation): 81.8°±8.8° and 145.0°±15.0°, respectively (P<0.005). There is an increase in the external ureteral temperature during laser activation, but ureteral thermal values decreased when saline flow was applied. Ureter thermal spread showed no difference between irrigated and nonirrigated subgroups. This is the first laser lithotripsy thermography study establishing the framework to evaluate the temperature

  13. Reversible, activity-dependent targeting of profilin to neuronal nuclei

    SciTech Connect

    Birbach, Andreas . E-mail: andreas.birbach@lbicr.lbg.ac.at; Verkuyl, J. Martin; Matus, Andrew . E-mail: aim@fmi.ch

    2006-07-15

    The actin cytoskeleton in pyramidal neurons plays a major role in activity-dependent processes underlying neuronal plasticity. The small actin-binding protein profilin shows NMDA receptor-dependent accumulation in dendritic spines, which is correlated with suppression of actin dynamics and long-term stabilization of synaptic morphology. Here we show that following NMDA receptor activation profilin also accumulates in the nucleus of hippocampal neurons via a process involving rearrangement of the actin cytoskeleton. This simultaneous targeting to dendritic spines and the cell nucleus suggests a novel mechanism of neuronal plasticity in which profilin both tags activated synapses and influences nuclear events.

  14. Colorimetric Determination of Pure Mg2+-dependent Phosphatidate Phosphatase Activity

    PubMed Central

    Havriluk, Tara; Lozy, Fred; Siniossoglou, Symeon; Carman, George M.

    2008-01-01

    The malachite green-molybdate reagent was used for a colorimetric assay of pure Mg2+-dependent phosphatidate phosphatase activity. This enzyme plays a major role in fat metabolism. Enzyme activity was linear with time and protein concentration, and with the concentration of water-soluble dioctanoyl phosphatidate. The colorimetric assay was used to examine enzyme inhibition by phenylglyoxal, propranolol, and dimethyl sulfoxide. Pure enzyme and a water-soluble phosphatidate substrate were required for the assay, which should be applicable to a well-defined large-scale screen of Mg2+-dependent phosphatidate phosphatase inhibitors (or activators). PMID:17910939

  15. Circadian Neuron Feedback Controls the Drosophila Sleep-Activity Profile

    PubMed Central

    Guo, Fang; Yu, Junwei; Jung, Hyung Jae; Abruzzi, Katharine C.; Luo, Weifei; Griffith, Leslie C.; Rosbash, Michael

    2016-01-01

    SUMMARY Little is known about the ability of Drosophila circadian neurons to promote sleep. We show here with optogenetic manipulations and video recording that a subset of dorsal clock neurons (DN1s) are potent sleep-promoting cells, releasing glutamate to directly inhibit key pacemaker neurons. These pacemakers promote morning arousal by activating these same DN1s, implying that there is a late-day feedback circuit to drive siesta and nighttime sleep. To address more plastic aspects of the sleep program, we used a novel calcium assay to monitor and compared the real-time DN1 activity of freely behaving males and females. It revealed a dramatic sexual dimorphism, which parallels the well-known difference in daytime sleep. DN1 activity is also enhanced by elevated temperature, consistent with its known effect on sleep. These new approaches indicate that the DN1s have a major impact on the fly sleep-wake profile and integrate environmental information with the circadian molecular program. PMID:27479324

  16. Phytochemical profiles and antioxidant activity of 27 cultivars of tea.

    PubMed

    Zeng, Liang; Luo, Liyong; Li, Hongjun; Liu, Ruihai

    2017-08-01

    Tea, rich in phytochemicals, has been suggested to have human health benefits. The phenolic profiles, antioxidant and antiproliferative activities of 27 tea cultivars were determined. Wide ranges of variation were found in analyzed cultivars for the contents of water-soluble phenolics (121.6-223.7 mg/g dry weight (DW)), total catechins (TC) (90.5-177.2 mg/g DW), antioxidant activities (PSC values 627.3-2332.3 μmol of vitamin C equiv/g DW, ORAC values (1865.1-3489.3 μmol of vitamin C equiv/g DW), CAA values (37.7-134.3 μmol of QE/g DW without PBS wash and 25.3-75.4 μmol of QE/g DW with PBS wash) and antiproliferative activity (53.0-90.8% at the concentration of 400 μg/mL extracts). The PSC, ORAC and CAA values were significantly correlated with phenolics, epicatechin gallate (ECG), CC and TC. Knowledge of specific differences among tea cultivars is important for breeding tea cultivars and gives sights to its potential application to promote health.

  17. Synthesis and Antidepressant Activity Profile of Some Novel Benzothiazole Derivatives.

    PubMed

    Demir Özkay, Ümide; Kaya, Ceren; Acar Çevik, Ulviye; Can, Özgür Devrim

    2017-09-07

    Within the scope of our new antidepressant drug development efforts, in this study, we synthesized eight novel benzothiazole derivatives 3a-3h. The chemical structures of the synthesized compounds were elucidated by spectroscopic methods. Test compounds were administered orally at a dose of 40 mg/kg to mice 24, 5 and 1 h before performing tail suspension, modified forced swimming, and activity cage tests. The obtained results showed that compounds 3c, 3d, 3f-3h reduced the immobility time of mice as assessed in the tail suspension test. Moreover, in the modified forced swimming tests, the same compounds significantly decreased the immobility, but increased the swimming frequencies of mice, without any alteration in the climbing frequencies. These results, similar to the results induced by the reference drug fluoxetine (20 mg/kg, po), indicated the antidepressant-like activities of the compounds 3c, 3d, 3f-3h. Owing to the fact that test compounds did not induce any significant alteration in the total number of spontaneous locomotor activities, the antidepressant-like effects of these derivatives seemed to be specific. In order to predict ADME parameters of the synthesized compounds 3a-3h, some physicochemical parameters were calculated. The ADME prediction study revealed that all synthesized compounds may possess good pharmacokinetic profiles.

  18. A comparison of cigarette smoking profiles in opioid-dependent pregnant patients receiving methadone or buprenorphine.

    PubMed

    Chisolm, Margaret S; Fitzsimons, Heather; Leoutsakos, Jeannie-Marie S; Acquavita, Shauna P; Heil, Sarah H; Wilson-Murphy, Molly; Tuten, Michelle; Kaltenbach, Karol; Martin, Peter R; Winklbaur, Bernadette; Jansson, Lauren M; Jones, Hendrée E

    2013-07-01

    Little is known about the relationship between cigarette smoking and agonist treatment in opioid-dependent pregnant patients. The objective of this study is to examine the extent to which cigarette smoking profiles differentially changed during the course of pregnancy in opioid-dependent patients receiving either double-blind methadone or buprenorphine. Patients were participants in the international, randomized controlled Maternal Opioid Treatment: Human Experimental Research (MOTHER) study. A sample of opioid-maintained pregnant patients (18-41 years old) with available smoking data who completed a multisite, double-blind, double-dummy, randomized controlled trial of methadone (n = 67) and buprenorphine (n = 57) between 2005 and 2008. Participants were compared on smoking variables based on opioid agonist treatment condition. Overall, 95% of the sample reported cigarette smoking at treatment entry. Participants in the two medication conditions were similar on pretreatment characteristics including smoking rates and daily cigarette amounts. Over the course of the pregnancy, no meaningful changes in cigarette smoking were observed for either medication condition. The fitted difference in change in adjusted cigarettes per day between the two conditions was small and nonsignificant (β = -0.08, SE = 0.05, p = .132). Results support high rates of smoking with little change during pregnancy among opioid-dependent patients, regardless of the type of agonist medication received. These findings are consistent with evidence that suggests nicotine effects, and interactions may be similar for buprenorphine compared with methadone. The outcomes further highlight that aggressive efforts are needed to reduce/eliminate smoking in opioid-dependent pregnant women.

  19. A Comparison of Cigarette Smoking Profiles in Opioid-Dependent Pregnant Patients Receiving Methadone or Buprenorphine

    PubMed Central

    2013-01-01

    Introduction: Little is known about the relationship between cigarette smoking and agonist treatment in opioid-dependent pregnant patients. The objective of this study is to examine the extent to which cigarette smoking profiles differentially changed during the course of pregnancy in opioid-dependent patients receiving either double-blind methadone or buprenorphine. Patients were participants in the international, randomized controlled Maternal Opioid Treatment: Human Experimental Research (MOTHER) study. Methods: A sample of opioid-maintained pregnant patients (18–41 years old) with available smoking data who completed a multisite, double-blind, double-dummy, randomized controlled trial of methadone (n = 67) and buprenorphine (n = 57) between 2005 and 2008. Participants were compared on smoking variables based on opioid agonist treatment condition. Results: Overall, 95% of the sample reported cigarette smoking at treatment entry. Participants in the two medication conditions were similar on pretreatment characteristics including smoking rates and daily cigarette amounts. Over the course of the pregnancy, no meaningful changes in cigarette smoking were observed for either medication condition. The fitted difference in change in adjusted cigarettes per day between the two conditions was small and nonsignificant (β = −0.08, SE = 0.05, p = .132). Conclusions: Results support high rates of smoking with little change during pregnancy among opioid-dependent patients, regardless of the type of agonist medication received. These findings are consistent with evidence that suggests nicotine effects, and interactions may be similar for buprenorphine compared with methadone. The outcomes further highlight that aggressive efforts are needed to reduce/eliminate smoking in opioid-dependent pregnant women. PMID:23288871

  20. Activity-dependent dendritic release of BDNF and biological consequences

    PubMed Central

    Kuczewski, Nicola; Porcher, Christophe; Lessmann, Volkmar; Medina, Igor; Gaiarsa, Jean-Luc

    2009-01-01

    Network construction and reorganization is modulated by the level and pattern of synaptic activity generated in the nervous system. During the past decades, neurotrophins, and in particular brain-derived neurotrophic factor (BDNF), have emerged as attractive candidates for linking synaptic activity and brain plasticity. Thus, neurotrophin expression and secretion are under the control of activity-dependent mechanisms and, besides their classical role in supporting neuronal survival neurotrophins, modulate nearly all key steps of network construction from neuronal migration to experience-dependent refinement of local connections. In this paper, we provide an overview of recent findings showing that BDNF can serve as a target-derived messenger for activity-dependent synaptic plasticity and development at the single cell level. PMID:19156544

  1. Muscle activity pattern dependent pain development and alleviation.

    PubMed

    Sjøgaard, Gisela; Søgaard, Karen

    2014-12-01

    Muscle activity is for decades considered to provide health benefits irrespectively of the muscle activity pattern performed and whether it is during e.g. sports, transportation, or occupational work tasks. Accordingly, the international recommendations for public health-promoting physical activity do not distinguish between occupational and leisure time physical activity. However, in this body of literature, attention has not been paid to the extensive documentation on occupational physical activity imposing a risk of impairment of health - in particular musculoskeletal health in terms of muscle pain. Focusing on muscle activity patterns and musculoskeletal health it is pertinent to elucidate the more specific aspects regarding exposure profiles and body regional pain. Static sustained muscle contraction for prolonged periods often occurs in the neck/shoulder area during occupational tasks and may underlie muscle pain development in spite of rather low relative muscle load. Causal mechanisms include a stereotype recruitment of low threshold motor units (activating type 1 muscle fibers) characterized by a lack of temporal as well as spatial variation in recruitment. In contrast during physical activities at leisure and sport the motor recruitment patterns are more dynamic including regularly relatively high muscle forces - also activating type 2 muscles fibers - as well as periods of full relaxation even of the type 1 muscle fibers. Such activity is unrelated to muscle pain development if adequate recovery is granted. However, delayed muscle soreness may develop following intensive eccentric muscle activity (e.g. down-hill skiing) with peak pain levels in thigh muscles 1-2 days after the exercise bout and a total recovery within 1 week. This acute pain profile is in contrast to the chronic muscle pain profile related to repetitive monotonous work tasks. The painful muscles show adverse functional, morphological, hormonal, as well as metabolic characteristics. Of

  2. IsomiR expression profiles in human lymphoblastoid cell lines exhibit population and gender dependencies.

    PubMed

    Loher, Phillipe; Londin, Eric R; Rigoutsos, Isidore

    2014-09-30

    For many years it was believed that each mature microRNA (miRNA) existed as a single entity with fixed endpoints and a 'static' and unchangeable primary sequence. However, recent evidence suggests that mature miRNAs are more 'dynamic' and that each miRNA precursor arm gives rise to multiple isoforms, the isomiRs. Here we report on our identification of numerous and abundant isomiRs in the lymphoblastoid cell lines (LCLs) of 452 men and women from five different population groups. Unexpectedly, we find that these isomiRs exhibit an expression profile that is population-dependent and gender-dependent. This is important as it indicates that the LCLs of each gender/population combination have their own unique collection of mature miRNA transcripts. Moreover, each identified isomiR has its own characteristic abundance that remains consistent across biological replicates indicating that these are not degradation products. The primary sequences of identified isomiRs differ from the known miRBase miRNA either at their 5´-endpoint (leads to a different 'seed' sequence and suggests a different targetome), their 3´-endpoint, or both simultaneously. Our analysis of Argonaute PAR-CLIP data from LCLs supports the association of many of these newly identified isomiRs with the Argonaute silencing complex and thus their functional roles through participation in the RNA interference pathway.

  3. IsomiR expression profiles in human lymphoblastoid cell lines exhibit population and gender dependencies

    PubMed Central

    Loher, Phillipe; Londin, Eric R.; Rigoutsos, Isidore

    2014-01-01

    For many years it was believed that each mature microRNA (miRNA) existed as a single entity with fixed endpoints and a ‘static’ and unchangeable primary sequence. However, recent evidence suggests that mature miRNAs are more ‘dynamic’ and that each miRNA precursor arm gives rise to multiple isoforms, the isomiRs. Here we report on our identification of numerous and abundant isomiRs in the lymphoblastoid cell lines (LCLs) of 452 men and women from five different population groups. Unexpectedly, we find that these isomiRs exhibit an expression profile that is population-dependent and gender-dependent. This is important as it indicates that the LCLs of each gender/population combination have their own unique collection of mature miRNA transcripts. Moreover, each identified isomiR has its own characteristic abundance that remains consistent across biological replicates indicating that these are not degradation products. The primary sequences of identified isomiRs differ from the known miRBase miRNA either at their 5´-endpoint (leads to a different ‘seed’ sequence and suggests a different targetome), their 3´-endpoint, or both simultaneously. Our analysis of Argonaute PAR-CLIP data from LCLs supports the association of many of these newly identified isomiRs with the Argonaute silencing complex and thus their functional roles through participation in the RNA interference pathway. PMID:25229428

  4. Cdk5-dependent phosphorylation of liprinα1 mediates neuronal activity-dependent synapse development.

    PubMed

    Huang, Huiqian; Lin, Xiaochen; Liang, Zhuoyi; Zhao, Teng; Du, Shengwang; Loy, Michael M T; Lai, Kwok-On; Fu, Amy K Y; Ip, Nancy Y

    2017-08-15

    The experience-dependent modulation of brain circuitry depends on dynamic changes in synaptic connections that are guided by neuronal activity. In particular, postsynaptic maturation requires changes in dendritic spine morphology, the targeting of postsynaptic proteins, and the insertion of synaptic neurotransmitter receptors. Thus, it is critical to understand how neuronal activity controls postsynaptic maturation. Here we report that the scaffold protein liprinα1 and its phosphorylation by cyclin-dependent kinase 5 (Cdk5) are critical for the maturation of excitatory synapses through regulation of the synaptic localization of the major postsynaptic organizer postsynaptic density (PSD)-95. Whereas Cdk5 phosphorylates liprinα1 at Thr701, this phosphorylation decreases in neurons in response to neuronal activity. Blockade of liprinα1 phosphorylation enhances the structural and functional maturation of excitatory synapses. Nanoscale superresolution imaging reveals that inhibition of liprinα1 phosphorylation increases the colocalization of liprinα1 with PSD-95. Furthermore, disruption of liprinα1 phosphorylation by a small interfering peptide, siLIP, promotes the synaptic localization of PSD-95 and enhances synaptic strength in vivo. Our findings collectively demonstrate that the Cdk5-dependent phosphorylation of liprinα1 is important for the postsynaptic organization during activity-dependent synapse development.

  5. Cdk5-dependent phosphorylation of liprinα1 mediates neuronal activity-dependent synapse development

    PubMed Central

    Huang, Huiqian; Lin, Xiaochen; Liang, Zhuoyi; Zhao, Teng; Du, Shengwang; Loy, Michael M. T.; Lai, Kwok-On; Fu, Amy K. Y.

    2017-01-01

    The experience-dependent modulation of brain circuitry depends on dynamic changes in synaptic connections that are guided by neuronal activity. In particular, postsynaptic maturation requires changes in dendritic spine morphology, the targeting of postsynaptic proteins, and the insertion of synaptic neurotransmitter receptors. Thus, it is critical to understand how neuronal activity controls postsynaptic maturation. Here we report that the scaffold protein liprinα1 and its phosphorylation by cyclin-dependent kinase 5 (Cdk5) are critical for the maturation of excitatory synapses through regulation of the synaptic localization of the major postsynaptic organizer postsynaptic density (PSD)-95. Whereas Cdk5 phosphorylates liprinα1 at Thr701, this phosphorylation decreases in neurons in response to neuronal activity. Blockade of liprinα1 phosphorylation enhances the structural and functional maturation of excitatory synapses. Nanoscale superresolution imaging reveals that inhibition of liprinα1 phosphorylation increases the colocalization of liprinα1 with PSD-95. Furthermore, disruption of liprinα1 phosphorylation by a small interfering peptide, siLIP, promotes the synaptic localization of PSD-95 and enhances synaptic strength in vivo. Our findings collectively demonstrate that the Cdk5-dependent phosphorylation of liprinα1 is important for the postsynaptic organization during activity-dependent synapse development. PMID:28760951

  6. Physiological Profile and Activity Pattern of Minor Gaelic Football Players.

    PubMed

    Cullen, Bryan D; Roantree, Mark T; McCarren, Andrew L; Kelly, David T; OʼConnor, Paul L; Hughes, Sarah M; Daly, Pat G; Moyna, Niall M

    2017-07-01

    Cullen, BD, Roantree, M, McCarren, A, Kelly, DT, O'Connor, PL, Hughes, SM, Daly, PG, and Moyna1, NM. Physiological profile and activity pattern of minor Gaelic football players. J Strength Cond Res 31(7): 1811-1820, 2017-The purpose of this study was to evaluate the physiological profile and activity pattern in club- and county-level under-18 (U-18) Gaelic football players relative to playing position. Participants (n = 85) were analyzed during 17 official 15-a-side matches using global positioning system technology (SPI Pro X II; GPSports Systems, Canberra, Australia) and heart rate (HR) telemetry. During the second part of this study, 63 participants underwent an incremental treadmill test to assess their maximal oxygen uptake (V[Combining Dot Above]o2max) and peak HR (HRmax). Players covered a mean distance of 5,774 ± 737 m during a full 60-minute match. The mean %HRmax and %V[Combining Dot Above]O2max observed during the match play were 81.6 ± 4.3% and 70.1 ± 7.75%, respectively. The playing level had no effect on the distance covered, player movement patterns, or %HRmax observed during match play. Midfield players covered significantly greater distance than defenders (p = 0.033). Playing position had no effect on %HRmax or the frequency of sprinting or high-intensity running during match play. The frequency of jogging, cruise running, striding (p = 0.000), and walking (p = 0.003) was greater in the midfield position than in the forward position. Time had a significant effect (F(1,39) = 33.512, p-value = 0.000, and (Equation is included in full-text article.)= 0.462) on distance covered and %HRmax, both of which showed a reduction between playing periods. Gaelic football is predominantly characterized by low-to-moderate intensity activity interspersed with periods of high-intensity running. The information provided may be used as a framework for coaches in the design and prescription of training strategies. Positional specific training may be warranted given

  7. Physicochemical signatures of nanoparticle-dependent complement activation

    SciTech Connect

    Thomas, Dennis G.; Chikkagoudar, Satish; Heredia-Langner, Alejandro; Tardiff, Mark F.; Xu, Zhixiang; Hourcade, Dennis; Pham, Christine; Lanza, Gregory M.; Weinberger, Kilian Q.; Baker, Nathan A.

    2014-03-21

    Nanoparticles are potentially powerful therapeutic tools that have the capacity to target drug payloads and imaging agents. However, some nanoparticles can activate complement, a branch of the innate immune system, and cause adverse side-effects. Recently, we developed an in vitro hemolytic assay protocol for measuring the nanoparticle-dependent complement activity of serum samples and applied this protocol to several nanoparticle formulations that differed in size, surface charge, and surface chemistry; quantifying the nanoparticle-dependent complement activity using a metric called Residual Hemolytic Activity (RHA). In the present work, we have used a decision tree learning algorithm to derive the rules for estimating nanoparticle-dependent complement response based on the data generated from the hemolytic assay studies. Our results indicate that physicochemical properties of nanoparticles, namely, size, polydispersity index, zeta potential, and mole percentage of the active surface ligand of a nanoparticle, can serve as good descriptors for prediction of nanoparticle-dependent complement activation in the decision tree modeling framework. The robustness and predictability of the model can be improved by training the model with additional data points that are uniformly distributed in the RHA/physicochemical descriptor space and by incorporating instability effects on nanoparticle physicochemical properties into the model.

  8. Distinct age and differentiation-state dependent metabolic profiles of oligodendrocytes under optimal and stress conditions.

    PubMed

    Rao, Vijayaraghava T S; Khan, Damla; Cui, Qiao-Ling; Fuh, Shih-Chieh; Hossain, Shireen; Almazan, Guillermina; Multhaup, Gerhard; Healy, Luke M; Kennedy, Timothy E; Antel, Jack P

    2017-01-01

    Within the microenvironment of multiple sclerosis lesions, oligodendrocytes are subject to metabolic stress reflecting effects of focal ischemia and inflammation. Previous studies have shown that under optimal conditions in vitro, the respiratory activity of human adult brain-derived oligodendrocytes is lower and more predominantly glycolytic compared to oligodendrocytes differentiated in vitro from post natal rat brain oligodendrocyte progenitor cells. In response to sub-lethal metabolic stress, adult human oligodendrocytes reduce overall energy production rate impacting the capacity to maintain myelination. Here, we directly compare the metabolic profiles of oligodendrocytes derived from adult rat brain with oligodendrocytes newly differentiated in vitro from oligodendrocyte progenitor cells obtained from the post natal rat brain, under both optimal culture and metabolic stress (low/no glucose) conditions. Oxygen consumption and extracellular acidification rates were measured using a Seahorse extracellular flux analyzer. Our findings indicate that under optimal conditions, adult rat oligodendrocytes preferentially use glycolysis whereas newly differentiated post natal rat oligodendrocytes, and the oligodendrocyte progenitor cells from which they are derived, mainly utilize oxidative phosphorylation to produce ATP. Metabolic stress increases the rate of ATP production via oxidative phosphorylation and significantly reduces glycolysis in adult oligodendrocytes. The rate of ATP production was relatively unchanged in newly differentiated post natal oligodendrocytes under these stress conditions, while it was significantly reduced in oligodendrocyte progenitor cells. Our study indicates that both age and maturation influence the metabolic profile under optimal and stressed conditions, emphasizing the need to consider these variables for in vitro studies that aim to model adult human disease.

  9. Psychopathological profile and prevalence of dual pathology on patients with alcoholic dependence undergoing outpatient treatment.

    PubMed

    García-Carretero, Miguel A; Novalbos-Ruiz, José P; Robles-Martínez, María; Jordán-Quintero, María A; O'Ferrall-González, Cristina

    2017-01-01

    Assess the prevalence of dual pathology in patients with alcohol dependence and describe the psychopathological profile of mental disorders, impulsiveness, ADHD presence and craving. It is a cross-sectional study about dual pathology, carried out on 102 patients undergoing outpatient treatment. The presence of dual pathology is established by means of the MINI-5 interview and the MCMI-III test; DSM-IV being used as the alcohol abuse criteria. Impulsiveness, ADHD presence, craving and quality of life were measured through SIS, ASRSv1, MACS and SF-36. The prevalence of dual pathology ranges from 45.1% to 80.4% according to MCMI-III and MINI-5, respectively. The most frequent pathologies are current major depressive episodes, followed by current generalized anxiety disorders, suicide risk and current dysthymia disorders; 73.2% of dual patients present a moderate and intense global score according to MACS, 56.1% got a meaningful score in impulsiveness according to SIS and 41.5% has highly consistent symptoms with ADHD. As regards quality of life, 53.7% of the sample had bad mental health. In the case of dual patients consuming other substances, 30% had a history of bipolar disorders and 10% had a high suicide risk. The prevalence of psychiatric comorbidity in patients with alcohol dependence undergoing outpatient treatment varies depending on the detection method, MINI being the one identifying a greater number of cases. More than half of dual patients present impulsive behavior, a bad mental health state and high craving levels. Special attention should be paid to dual patients consuming other substances.

  10. Niche-Dependent Gene Expression Profile of Intratumoral Heterogeneous Ovarian Cancer Stem Cell Populations

    PubMed Central

    Abelson, Sagi; Shamai, Yeela; Berger, Liron; Skorecki, Karl; Tzukerman, Maty

    2013-01-01

    Intratumoral heterogeneity challenges existing paradigms for anti-cancer therapy. We have previously demonstrated that the human embryonic stem cells (hESC)-derived cellular microenvironment in immunocompromised mice, enables functional distinction of heterogeneous tumor cells, including cells which do not grow into a tumor in a conventional direct tumor xenograft platform. We have identified and characterized six cancer cell subpopulations each clonally expanded from a single cell, derived from human ovarian clear cell carcinoma of a single tumor, to demonstrate striking intratumoral phenotypic heterogeneity that is dynamically dependent on the tumor growth microenvironment. These cancer cell subpopulations, characterized as cancer stem cell subpopulations, faithfully recapitulate the full spectrum of histological phenotypic heterogeneity known for human ovarian clear cell carcinoma. Each of the six subpopulations displays a different level of morphologic and tumorigenic differentiation wherein growth in the hESC-derived microenvironment favors growth of CD44+/aldehyde dehydrogenase positive pockets of self-renewing cells that sustain tumor growth through a process of tumorigenic differentiation into CD44-/aldehyde dehydrogenase negative derivatives. Strikingly, these derivative cells display microenvironment-dependent plasticity with the capacity to restore self-renewal markers and CD44 expression. In the current study, we delineate the distinct gene expression and epigenetic profiles of two such subpopulations, representing extremes of phenotypic heterogeneity in terms of niche-dependent self-renewal and tumorigenic differentiation. By combining Gene Set Enrichment, Gene Ontology and Pathway-focused array analyses with methylation status, we propose a suite of robust differences in tumor self-renewal and differentiation pathways that underlie the striking intratumoral phenotypic heterogeneity which characterize this and other solid tumor malignancies. PMID

  11. Profiles.

    ERIC Educational Resources Information Center

    School Arts, 1979

    1979-01-01

    Profiles seven Black, Native American, and Chicano artists and art teachers: Hale A. Woodruff, Allan Houser, Luis Jimenez, Betrand D. Phillips, James E. Pate, I, and Fernando Navarro. This article is part of a theme issue on multicultural art. (SJL)

  12. Profiles.

    ERIC Educational Resources Information Center

    School Arts, 1979

    1979-01-01

    Profiles seven Black, Native American, and Chicano artists and art teachers: Hale A. Woodruff, Allan Houser, Luis Jimenez, Betrand D. Phillips, James E. Pate, I, and Fernando Navarro. This article is part of a theme issue on multicultural art. (SJL)

  13. DNA methylation profiling of esophageal adenocarcinoma using Methylation Ligation-dependent Macroarray (MLM).

    PubMed

    Guilleret, Isabelle; Losi, Lorena; Chelbi, Sonia T; Fonda, Sergio; Bougel, Stéphanie; Saponaro, Sara; Gozzi, Gaia; Alberti, Loredana; Braunschweig, Richard; Benhattar, Jean

    2016-10-14

    Most types of cancer cells are characterized by aberrant methylation of promoter genes. In this study, we described a rapid, reproducible, and relatively inexpensive approach allowing the detection of multiple human methylated promoter genes from many tissue samples, without the need of bisulfite conversion. The Methylation Ligation-dependent Macroarray (MLM), an array-based analysis, was designed in order to measure methylation levels of 58 genes previously described as putative biomarkers of cancer. The performance of the design was proven by screening the methylation profile of DNA from esophageal cell lines, as well as microdissected formalin-fixed and paraffin-embedded (FFPE) tissues from esophageal adenocarcinoma (EAC). Using the MLM approach, we identified 32 (55%) hypermethylated promoters in EAC, and not or rarely methylated in normal tissues. Among them, 21promoters were found aberrantly methylated in more than half of tumors. Moreover, seven of them (ADAMTS18, APC, DKK2, FOXL2, GPX3, TIMP3 and WIF1) were found aberrantly methylated in all or almost all the tumor samples, suggesting an important role for these genes in EAC. In addition, dysregulation of the Wnt pathway with hypermethylation of several Wnt antagonist genes was frequently observed. MLM revealed a homogeneous pattern of methylation for a majority of tumors which were associated with an advanced stage at presentation and a poor prognosis. Interestingly, the few tumors presenting less methylation changes had a lower pathological stage. In conclusion, this study demonstrated the feasibility and accuracy of MLM for DNA methylation profiling of FFPE tissue samples. Copyright © 2016 Elsevier Inc. All rights reserved.

  14. Stage-dependent gene expression profiles during natural killer cell development.

    PubMed

    Kang, Hyung-Sik; Kim, Eun-Mi; Lee, Sanggyu; Yoon, Suk-Ran; Kawamura, Toshihiko; Lee, Young-Cheol; Kim, Sangsoo; Myung, Pyung-Keun; Wang, San Ming; Choi, Inpyo

    2005-11-01

    Natural killer (NK) cells develop from hematopoietic stem cells (HSCs) in the bone marrow. To understand the molecular regulation of NK cell development, serial analysis of gene expression (SAGE) was applied to HSCs, NK precursor (pNK) cells, and mature NK cells (mNK) cultured without or with OP9 stromal cells. From 170,464 total individual tags from four SAGE libraries, 35,385 unique genes were identified. A set of genes was expressed in a stage-specific manner: 15 genes in HSCs, 30 genes in pNK cells, and 27 genes in mNK cells. Among them, lipoprotein lipase induced NK cell maturation and cytotoxic activity. Identification of genome-wide profiles of gene expression in different stages of NK cell development affords us a fundamental basis for defining the molecular network during NK cell development.

  15. Profiles of Childhood Trauma in Patients with Alcohol Dependence and Their Associations with Addiction-Related Problems.

    PubMed

    Lotzin, Annett; Haupt, Lena; von Schönfels, Julia; Wingenfeld, Katja; Schäfer, Ingo

    2016-03-01

    The high occurrence of childhood trauma in individuals with alcohol dependence is well-recognized. Nevertheless, researchers have rarely studied which types of childhood trauma often co-occur and how these combinations of different types and severities of childhood trauma are related to the patients' current addiction-related problems. We aimed to identify childhood trauma profiles in patients with alcohol dependence and examined relations of these trauma profiles with the patients' current addiction-related problems. In 347 alcohol-dependent patients, 5 types of childhood trauma (sexual abuse, physical abuse, emotional abuse, emotional neglect, and physical neglect) were measured using the Childhood Trauma Questionnaire. Childhood trauma profiles were identified using cluster analysis. The patients' current severity of addiction-related problems was assessed using the European Addiction Severity Index. We identified 6 profiles that comprised different types and severities of childhood trauma. The patients' trauma profiles predicted the severity of addiction-related problems in the domains of psychiatric symptoms, family relationships, social relationships, and drug use. Childhood trauma profiles may provide more useful information about the patient's risk of current addiction-related problems than the common distinction between traumatized versus nontraumatized patients. Copyright © 2016 by the Research Society on Alcoholism.

  16. On deriving the accurate aerosol extinction profiles in the troposphere and lower stratosphere using the range dependent scattering ratio

    NASA Astrophysics Data System (ADS)

    Satyanarayana, M. V.; Radhakrishnan, S. R.; Mahadevanpillai, V. P.; Krishnakumar, V.

    2008-12-01

    Lidar has proven to be an effective instrument for obtaining high resolution profiles of atmospheric aerosols. Deriving the optical properties of aerosols from the experimentally obtained lidar data is one of the most interesting and challenging task for the atmospheric scientists. A few methods had been developed so far, to obtain the quantitative profiles of extinction and backscattering coefficient of aerosols from the pulsed backscattering lidar measurements. Most of the existing inversion methods assume a range independent value for the scattering ratio for inverting the lidar signal even though it is known that the scattering ratio depends on the nature of aerosols and as such range dependent. We used a modified Klett's method for the inversion of lidar signal that uses range dependent scattering ratio (s) for the characterization of atmospheric aerosols. This method provides the constants k and s for all the altitude regions of the atmosphere and leads to derive the aerosol extinction profile for the lidar data. In this paper we made a study on the errors involved in the extinction profiles derived using the range dependent scattering ratio and discuss the approach in this regard to obtain the accurate extinction profiles.

  17. Substrate-Competitive Activity-Based Profiling of Ester Prodrug Activating Enzymes.

    PubMed

    Xu, Hao; Majmudar, Jaimeen D; Davda, Dahvid; Ghanakota, Phani; Kim, Ki H; Carlson, Heather A; Showalter, Hollis D; Martin, Brent R; Amidon, Gordon L

    2015-09-08

    Understanding the mechanistic basis of prodrug delivery and activation is critical for establishing species-specific prodrug sensitivities necessary for evaluating preclinical animal models and potential drug-drug interactions. Despite significant adoption of prodrug methodologies for enhanced pharmacokinetics, functional annotation of prodrug activating enzymes is laborious and often unaddressed. Activity-based protein profiling (ABPP) describes an emerging chemoproteomic approach to assay active site occupancy within a mechanistically similar enzyme class in native proteomes. The serine hydrolase enzyme family is broadly reactive with reporter-linked fluorophosphonates, which have shown to provide a mechanism-based covalent labeling strategy to assay the activation state and active site occupancy of cellular serine amidases, esterases, and thioesterases. Here we describe a modified ABPP approach using direct substrate competition to identify activating enzymes for an ethyl ester prodrug, the influenza neuraminidase inhibitor oseltamivir. Substrate-competitive ABPP analysis identified carboxylesterase 1 (CES1) as an oseltamivir-activating enzyme in intestinal cell homogenates. Saturating concentrations of oseltamivir lead to a four-fold reduction in the observed rate constant for CES1 inactivation by fluorophosphonates. WWL50, a reported carbamate inhibitor of mouse CES1, blocked oseltamivir hydrolysis activity in human cell homogenates, confirming CES1 is the primary prodrug activating enzyme for oseltamivir in human liver and intestinal cell lines. The related carbamate inhibitor WWL79 inhibited mouse but not human CES1, providing a series of probes for analyzing prodrug activation mechanisms in different preclinical models. Overall, we present a substrate-competitive activity-based profiling approach for broadly surveying candidate prodrug hydrolyzing enzymes and outline the kinetic parameters for activating enzyme discovery, ester prodrug design, and

  18. Evolving Hox Activity Profiles Govern Diversity in Locomotor Systems

    PubMed Central

    Jung, Heekyung; Mazzoni, Esteban O.; Soshnikova, Natalia; Hanley, Olivia; Venkatesh, Byrappa; Duboule, Denis; Dasen, Jeremy S.

    2014-01-01

    Summary The emergence of limb-driven locomotor behaviors was a key event in the evolution of vertebrates and fostered the transition from aquatic to terrestrial life. We show that the generation of limb-projecting lateral motor column (LMC) neurons in mice relies on a transcriptional autoregulatory module initiated via transient activity of multiple genes within the HoxA and HoxC clusters. Repression of this module at thoracic levels restricts expression of LMC determinants, thus dictating LMC position relative to the limbs. This suppression is mediated by a key regulatory domain that is specifically found in the Hoxc9 proteins of appendage-bearing vertebrates. The profile of Hoxc9 expression inversely correlates with LMC position in land vertebrates, and likely accounts for the absence of LMC neurons in limbless species such as snakes. Thus, modulation of both Hoxc9 protein function and Hoxc9 gene expression likely contributed to evolutionary transitions between undulatory and ambulatory motor circuit connectivity programs. PMID:24746670

  19. Target identification with quantitative activity based protein profiling (ABPP).

    PubMed

    Chen, Xiao; Wong, Yin Kwan; Wang, Jigang; Zhang, Jianbin; Lee, Yew-Mun; Shen, Han-Ming; Lin, Qingsong; Hua, Zi-Chun

    2017-02-01

    As many small bioactive molecules fulfill their functions through interacting with protein targets, the identification of such targets is crucial in understanding their mechanisms of action (MOA) and side effects. With technological advancements in target identification, it has become possible to accurately and comprehensively study the MOA and side effects of small molecules. While small molecules with therapeutic potential were derived solely from nature in the past, the remodeling and synthesis of such molecules have now been made possible. Presently, while some small molecules have seen successful application as drugs, the majority remain undeveloped, requiring further understanding of their MOA and side effects to fully tap into their potential. Given the typical promiscuity of many small molecules and the complexity of the cellular proteome, a high-flux and high-accuracy method is necessary. While affinity chromatography approaches combined with MS have had successes in target identification, limitations associated with nonspecific results remain. To overcome these complications, quantitative chemical proteomics approaches have been developed including metabolic labeling, chemical labeling, and label-free methods. These new approaches are adopted in conjunction with activity-based protein profiling (ABPP), allowing for a rapid process and accurate results. This review will briefly introduce the principles involved in ABPP, then summarize current advances in quantitative chemical proteomics approaches as well as illustrate with examples how ABPP coupled with quantitative chemical proteomics has been used to detect the targets of drugs and other bioactive small molecules including natural products.

  20. Age-dependent alterations of the NMDA receptor developmental profile and adult behavior in postnatally ketamine-treated mice.

    PubMed

    Lecointre, Maryline; Vézier, Claire; Bénard, Magalie; Ramdani, Yasmina; Dupré, Nicolas; Brasse-Lagnel, Carole; Henry, Vincent J; Roy, Vincent; Marret, Stéphane; Gonzalez, Bruno J; Jégou, Sylvie; Leroux-Nicollet, Isabelle

    2015-03-01

    Ketamine is a NMDA receptor (NMDAR) antagonist used in pediatric anesthesia. Given the role of glutamatergic signaling during brain maturation, we studied the effects of a single ketamine injection (40 mg/kg s.c) in mouse neonates depending on postnatal age at injection (P2, P5, or P10) on cortical NMDAR subunits expression and association with Membrane-Associated Guanylate Kinases PSD95 and SAP102. The effects of ketamine injection at P2, P5, or P10 on motor activity were compared in adulthood. Ketamine increased GluN2A and GluN2B mRNA levels in P2-treated mice without change in proteins, while it decreased GluN2B protein in P10-treated mice without change in mRNA. Ketamine reduced GluN2A mRNA and protein levels in P5-treated mice without change in GluN2B and GluN1. Ketamine affected the GluN2A/PSD95 association regardless of the age at injection, while GluN2B/PSD95 association was enhanced only in P5-treated mice. Microdissection of ketamine-treated mouse cortex showed a decrease in GluN2A mRNA level in superficial layers (I-IV) and an increase in all subunit expressions in deep layers (V-VI) in P5- and P10-treated mice, respectively. Our data suggest that ketamine impairs cortical NMDAR subunit developmental profile and delays the synaptic targeting of GluN2A-enriched NMDAR. Ketamine injection at P2 or P10 resulted in hyperlocomotion in adult male mice in an open field, without change in females. Voluntary running-wheel exercise showed age- and sex-dependent alterations of the mouse activity, especially during the dark phase. Overall, a single neonatal ketamine exposure led to short-term NMDAR cortical developmental profile impairments and long-term motor activity alterations persisting in adulthood. © 2014 Wiley Periodicals, Inc.

  1. Plasma Biomarker Profiles Differ Depending on Breast Cancer Subtype but RANTES is Consistently Increased

    SciTech Connect

    Gonzales, Rachel M.; Daly, Don S.; Tan, Ruimin; Marks, Jeffrey R.; Zangar, Richard C.

    2011-07-01

    Background: Current biomarkers for breast cancer have little potential for detection. We determined if breast cancer subtypes influence circulating protein biomarkers. Methods: A sandwich-ELISA microarray platform was used to evaluate 23 candidate biomarkers in plasma samples that were obtained from subjects with either benign breast disease or invasive breast cancer. All plasma samples were collected at the time of biopsy, after a referral due to a suspicious screen (e.g., mammography). Cancer samples were evaluated based on breast cancer subtypes, as defined by the HER2 and estrogen receptor statuses. Results: Ten proteins were statistically altered in at least one breast cancer subtype, including four epidermal growth factor receptor ligands, two matrix metalloproteases, two cytokines, and two angiogenic factors. Only one cytokine, RANTES, was significantly increased (P<0.01 for each analysis) in all four subtypes, with areas under receiver operating characteristic curves (AUC) that ranged from 0.76 to 0.82, depending on cancer subtype. The best AUC values were observed for analyses that combined data from multiple biomarkers, with values ranging from 0.70 to 0.99, depending on the cancer subtype. Although the results for RANTES are consistent with previous publications, the multi-assay results need to be validated in independent sample sets. Conclusions: Different breast cancer subtypes produce distinct biomarker profiles, and circulating protein biomarkers have potential to differentiate between true and false positive screens for breast cancer. Impact: Subtype-specific biomarker panels may be useful for detecting breast cancer or as an adjunct assay to improve the accuracy of current screening methods.

  2. Safety Profile of Oral Iron Chelator Deferiprone in Chinese Children with Transfusion-Dependent Thalassaemia.

    PubMed

    Botzenhardt, Sebastian; Sing, Chor W; Wong, Ian C K; Chan, Godfrey Chi-Fung; Wong, Lisa Y L; Felisi, Mariagrazia; Rascher, Wolfgang; Ceci, Adriana; Neubert, Antje

    2016-01-01

    There is a lack of knowledge regarding the incidence of serious adverse drug reactions (ADR) to the oral iron chelator deferiprone in Chinese children with transfusion-dependent thalassaemia. In this retrospective population-based cohort study, paediatric thalassaemia patients in Hong Kong were screened for serious and medically important adverse events related to deferiprone therapy using diagnosis codes, laboratory data and hospital admissions. Potential ADRs were assessed by reviewing concomitant medications, diagnoses and laboratory data and evaluated using standardised causality assessment. Eighty-seven patients contributing 169.8 person-years were included. Thirty ADRs were identified in 21 patients. Most ADRs (56.0%) occurred in the first three months of therapy. Neutropenia occurred in 11 patients (12.6%; incidence rate 6.5 per 100 patient-years) and severe neutropenia (agranulocytosis) was observed in 5 patients (5.7%, incidence rate 2.9 per 100 patient-years). Other identified ADRs involve severe arthropathy, elevated liver enzymes and mild thrombocytopenia. In conclusion, the safety profile of DFP therapy in Chinese children suffering from transfusion-dependent thalassaemia is in line with previous studies of non-Chinese children. However, unlike previous studies, we observed a relatively high incidence of agranulocytosis and neutropenia in patients with simultaneous combined therapy. Hence close monitoring for white blood cell counts is advised in Chinese children under combined iron chelation therapy. Further prospective clinical and pharmacogenetic studies are required to better evaluate this important safety signal. • Half of the identified ADRs related to deferiprone therapy occurred during the first three months of treatment. • A relatively high incidence of agranulocytosis and neutropenia. Hence close monitoring for white blood cell counts is advised in Chinese children under combined iron chelation therapy.

  3. Chloride dependence of hyperpolarization-activated chloride channel gates

    PubMed Central

    Pusch, Michael; Jordt, Sven-Eric; Stein, Valentin; Jentsch, Thomas J

    1999-01-01

    ClC proteins are a class of voltage-dependent Cl− channels with several members mutated in human diseases. The prototype ClC-0 Torpedo channel is a dimeric protein; each subunit forms a pore that can gate independently from the other one. A common slower gating mechanism acts on both pores simultaneously; slow gating activates ClC-0 at hyperpolarized voltages. The ClC-2 Cl− channel is also activated by hyperpolarization, as are some ClC-1 mutants (e.g. D136G) and wild-type (WT) ClC-1 at certain pH values.We studied the dependence on internal Cl− ([Cl−]i) of the hyperpolarization-activated gates of several ClC channels (WT ClC-0, ClC-0 mutant P522G, ClC-1 mutant D136G and an N-terminal deletion mutant of ClC-2), by patch clamping channels expressed in Xenopus oocytes.With all these channels, reducing [Cl−]i shifted activation to more negative voltages and reduced the maximal activation at most negative voltages.We also investigated the external halide dependence of WT ClC-2 using two-electrode voltage-clamp recording. Reducing external Cl− ([Cl−]o) activated ClC-2 currents. Replacing [Cl−]o by the less permeant Br− reduced channel activity and accelerated deactivation.Gating of the ClC-2 mutant K566Q in normal [Cl−]o resembled that of WT ClC-2 in low [Cl−]o, i.e. channels had a considerable open probability (Po) at resting membrane potential. Substituting external Cl− by Br− or I− led to a decrease in Po.The [Cl−]i dependence of the hyperpolarization-activated gates of various ClC channels suggests a similar gating mechanism, and raises the possibility that the gating charge for the hyperpolarization-activated gate is provided by Cl−.The external halide dependence of hyperpolarization-activated gating of ClC-2 suggests that it is mediated or modulated by anions as in other ClC channels. In contrast to the depolarization-activated fast gates of ClC-0 and ClC-1, the absence of Cl− favours channel opening. Lysine 556 may be important

  4. Illustrating the Effect of pH on Enzyme Activity Using Gibbs Energy Profiles

    ERIC Educational Resources Information Center

    Bearne, Stephen L.

    2014-01-01

    Gibbs energy profiles provide students with a visual representation of the energy changes that occur during enzyme catalysis, making such profiles useful as teaching and learning tools. Traditional kinetic topics, such as the effect of pH on enzyme activity, are often not discussed in terms of Gibbs energy profiles. Herein, the symbolism of Gibbs…

  5. Illustrating the Effect of pH on Enzyme Activity Using Gibbs Energy Profiles

    ERIC Educational Resources Information Center

    Bearne, Stephen L.

    2014-01-01

    Gibbs energy profiles provide students with a visual representation of the energy changes that occur during enzyme catalysis, making such profiles useful as teaching and learning tools. Traditional kinetic topics, such as the effect of pH on enzyme activity, are often not discussed in terms of Gibbs energy profiles. Herein, the symbolism of Gibbs…

  6. Etoposide Induces ATM-Dependent Mitochondrial Biogenesis through AMPK Activation

    PubMed Central

    Lyu, Yi Lisa; Liu, Leroy F.; Qi, Haiyan

    2008-01-01

    Background DNA damage such as double-stranded DNA breaks (DSBs) has been reported to stimulate mitochondrial biogenesis. However, the underlying mechanism is poorly understood. The major player in response to DSBs is ATM (ataxia telangiectasia mutated). Upon sensing DSBs, ATM is activated through autophosphorylation and phosphorylates a number of substrates for DNA repair, cell cycle regulation and apoptosis. ATM has been reported to phosphorylate the α subunit of AMP-activated protein kinase (AMPK), which senses AMP/ATP ratio in cells, and can be activated by upstream kinases. Here we provide evidence for a novel role of ATM in mitochondrial biogenesis through AMPK activation in response to etoposide-induced DNA damage. Methodology/Principal Findings Three pairs of human ATM+ and ATM- cells were employed. Cells treated with etoposide exhibited an ATM-dependent increase in mitochondrial mass as measured by 10-N-Nonyl-Acridine Orange and MitoTracker Green FM staining, as well as an increase in mitochondrial DNA content. In addition, the expression of several known mitochondrial biogenesis regulators such as the major mitochondrial transcription factor NRF-1, PGC-1α and TFAM was also elevated in response to etoposide treatment as monitored by RT-PCR. Three pieces of evidence suggest that etoposide-induced mitochondrial biogenesis is due to ATM-dependent activation of AMPK. First, etoposide induced ATM-dependent phosphorylation of AMPK α subunit at Thr172, indicative of AMPK activation. Second, inhibition of AMPK blocked etoposide-induced mitochondrial biogenesis. Third, activation of AMPK by AICAR (an AMP analogue) stimulated mitochondrial biogenesis in an ATM-dependent manner, suggesting that ATM may be an upstream kinase of AMPK in the mitochondrial biogenesis pathway. Conclusions/Significance These results suggest that activation of ATM by etoposide can lead to mitochondrial biogenesis through AMPK activation. We propose that ATM-dependent mitochondrial

  7. RNA-dependent DNA-polymerase activity in human milk.

    PubMed

    McFarlane, E S; Ryan, S M; Mann, E

    1975-08-01

    A simple method is described for testing milk specimens from nursing mothers for the presence of RNA-dependent DNA-polymerase activity. Positive results were obtained in five of 137 women (3.6%) without a family history of breast cancer, and in six of 31 women (19.3%) with a family history of breast cancer.

  8. Unconscious Semantic Activation Depends on Feature-Specific Attention Allocation

    ERIC Educational Resources Information Center

    Spruyt, Adriaan; De Houwer, Jan; Everaert, Tom; Hermans, Dirk

    2012-01-01

    We examined whether semantic activation by subliminally presented stimuli is dependent upon the extent to which participants assign attention to specific semantic stimulus features and stimulus dimensions. Participants pronounced visible target words that were preceded by briefly presented, masked prime words. Both affective and non-affective…

  9. Unconscious Semantic Activation Depends on Feature-Specific Attention Allocation

    ERIC Educational Resources Information Center

    Spruyt, Adriaan; De Houwer, Jan; Everaert, Tom; Hermans, Dirk

    2012-01-01

    We examined whether semantic activation by subliminally presented stimuli is dependent upon the extent to which participants assign attention to specific semantic stimulus features and stimulus dimensions. Participants pronounced visible target words that were preceded by briefly presented, masked prime words. Both affective and non-affective…

  10. Dysregulation of Macrophage Activation Profiles by Engineered Nanoparticles

    PubMed Central

    Kodali, Vamsi; Littke, Matthew H.; Tilton, Susan C.; Teeguarden, Justin G.; Shi, Liang; Frevert, Charles W.; Wang, Wei; Pounds, Joel G.; Thrall, Brian D.

    2013-01-01

    Although the potential human health impacts from exposure to engineered nanoparticles (ENPs) are uncertain, past epidemiological studies have established correlations between exposure to ambient air pollution particulates and the incidence of pneumonia and lung infections. Using amorphous silica and superparamagnetic iron oxide (SPIO) as model high production volume ENPs, we examined how macrophage activation by bacterial lipopolysaccharide (LPS) or the lung pathogen Streptococcus pneumoniae is altered by ENP pre-treatment. Neither silica nor SPIO treatment elicited direct cytotoxic or pro-inflammatory effects in bone marrow-derived macrophages. However, pre-treatment of macrophages with SPIO caused extensive reprogramming of nearly 500 genes regulated in response to LPS challenge, hallmarked by exaggerated activation of oxidative stress response pathways and suppressed activation of both pro- and anti-inflammatory pathways. Silica pre-treatment altered regulation of only 67 genes, but there was strong correlation with gene sets affected by SPIO. Macrophages exposed to SPIO displayed a phenotype suggesting an impaired ability to transition from a M1 to M2-like activation state, characterized by suppressed IL-10 induction, enhanced TNFα production, and diminished phagocytic activity toward S. pneumoniae. Studies in macrophages deficient in scavenger receptor A (SR-A) showed SR-A participates in cell uptake of both the ENPs and S. pneumonia, and co-regulates the anti-inflammatory IL-10 pathway. Thus, mechanisms for dysregulation of innate immunity exist by virtue that common receptor recognition pathways are used by some ENPs and pathogenic bacteria, although the extent of transcriptional reprogramming of macrophage function depends on the physicochemical properties of the ENP after internalization. Our results also illustrate that biological effects of ENPs may be indirectly manifested only after challenging normal cell function. Nanotoxicology screening strategies

  11. Dysregulation of Macrophage Activation Profiles by Engineered Nanoparticles

    SciTech Connect

    Kodali, Vamsi; Littke, Matthew H.; Tilton, Susan C.; Teeguarden, Justin G.; Shi, Liang; Frevert, Charles W.; Wang, Wei; Pounds, Joel G.; Thrall, Brian D.

    2013-08-27

    Although the potential human health impacts from exposure to engineered nanoparticles (ENPs) are uncertain, past epidemiological studies have established correlations between exposure to ambient air pollution particulates and the incidence of pneumonia and lung infections. Using amorphous silica and superparamagnetic iron oxide (SPIO) as model high production volume ENPs, we examined how macrophage activation by bacterial lipopolysaccharide (LPS) or the lung pathogen Streptococcus pneumoniae is altered by ENP pretreatment. Neither silica nor SPIO treatment elicited direct cytotoxic or pro-inflammatory effects in bone marrow-derived macrophages. However, pretreatment of macrophages with SPIO caused extensive reprogramming of nearly 500 genes regulated in response to LPS challenge, hallmarked by exaggerated activation of oxidative stress response pathways and suppressed activation of both pro- and anti-inflammatory pathways. Silica pretreatment altered regulation of only 67 genes, but there was strong correlation with gene sets affected by SPIO. Macrophages exposed to SPIO displayed a phenotype suggesting an impaired ability to transition from an M1 to M2-like activation state, characterized by suppressed IL-10 induction, enhanced TNFα production, and diminished phagocytic activity toward S. pneumoniae. Studies in macrophages deficient in scavenger receptor A (SR-A) showed SR-A participates in cell uptake of both the ENPs and S. pneumonia and co-regulates the anti-inflammatory IL-10 pathway. Thus, mechanisms for dysregulation of innate immunity exist by virtue that common receptor recognition pathways are used by some ENPs and pathogenic bacteria, although the extent of transcriptional reprogramming of macrophage function depends on the physicochemical properties of the ENP after internalization. Our results also illustrate that biological effects of ENPs may be indirectly manifested only after challenging normal cell function. Finally, nanotoxicology screening

  12. Activity profile of young soccer players during actual match play.

    PubMed

    Castagna, Carlo; D'Ottavio, Stefano; Abt, Grant

    2003-11-01

    This study investigated the activity profile of young soccer players (mean age 11.8 +/- 0.6 years; N = 12) with the aim of providing information for the development of training strategies. Data for movements of each player were obtained using 2 cameras that aimed at the subject throughout the match (Play Controller, Phromos, Italy). Encoders transmitted camera movements to a computer. The collected signals were then converted into distances, times, and speed attained at arbitrarily selected match categories. Players were monitored during official 11 vs. 11 matches (N = 12), with each match consisting of 2 halves, each lasting 30 minutes played on a regular soccer pitch. Mean total match distance amounted to 6,175 +/- 318 m. During the second half, players covered 5.5% less distance (p > 0.05). At speeds between 13.1 and 18.0 km.h(-1), players covered 12% less distance during the second half (p < 0.05). Players stood still 11% of the total time played (3,789 +/- 109 seconds). At speeds faster than 18 km.h(-1), players performed 33 +/- 4 bouts during match play, with a mean time length of 2.3 +/- 0.6 seconds per bout. Mean time interval between two successive maximal sprint bouts was 118.5 +/- 20.5 seconds. Players stood still longer during the course of the second half (229 +/- 76 seconds vs. 173 +/- 61 seconds, p < 0.05). Players tended to play in small areas of the football pitch and spent 9% of the total match time at high intensity. In order to promote a more active space coverage of the young player, at least for the age considered in the present study, the dimensions of the football pitch and the number of players should be reduced.

  13. Kickboxing review: anthropometric, psychophysiological and activity profiles and injury epidemiology

    PubMed Central

    Chaabene, H; Miarka, B; Franchini, E; Chamari, K; Cheour, F

    2017-01-01

    Kickboxing is one of the modern combat sports. The psychophysiological demands of a kickboxing competition require athletes to achieve high thresholds of several aspects of physical fitness. The aim of the current review is to critically analyse and appraise the kickboxer’s anthropometric, physiological, physical and psychological attributes with the activity profile and injury epidemiology in order to provide practical recommendations for training as well as new areas of scientific research. The available information shows that both amateur and elite-level male kickboxers are characterized by a higher proportion of mesomorphy with a well-developed muscle mass and low body fat percentage. While there is some variation in the maximum oxygen uptake of kickboxers, moderate to high cardio-respiratory levels are reported for these athletes. Regardless of kickboxers’ level, a high peak and mean anaerobic power output were reported. High-level kickboxing performance also requires well-developed muscle power in both the upper and lower limbs. Psychological factors contribute to success that requires high levels of self-confidence, motivation, dispositional hope and optimism, mental toughness/resiliency, and adaptive perfectionism. Psychological attributes also distinguished successful from less successful kickboxers. The activity-to-rest ratio was higher in elite (1:1) than both amateur and national-level (from 1:2 to 1:5) kickboxers, with no significant differences between rounds (round 1=1:4, and rounds 2 and 3=1:5) as well as between winners and losers in amateur and national-level simulated combats. These particular psychophysiological characteristics and performance aspects of kickboxers influence performance and could serve as guidance for training. Finally, kickboxing is characterized by chronic repetitive head trauma, which causes hypopituitarism due to traumatic brain injury (TBI). Future investigations into the physical, physiological and psychological

  14. Kickboxing review: anthropometric, psychophysiological and activity profiles and injury epidemiology.

    PubMed

    Slimani, M; Chaabene, H; Miarka, B; Franchini, E; Chamari, K; Cheour, F

    2017-06-01

    Kickboxing is one of the modern combat sports. The psychophysiological demands of a kickboxing competition require athletes to achieve high thresholds of several aspects of physical fitness. The aim of the current review is to critically analyse and appraise the kickboxer's anthropometric, physiological, physical and psychological attributes with the activity profile and injury epidemiology in order to provide practical recommendations for training as well as new areas of scientific research. The available information shows that both amateur and elite-level male kickboxers are characterized by a higher proportion of mesomorphy with a well-developed muscle mass and low body fat percentage. While there is some variation in the maximum oxygen uptake of kickboxers, moderate to high cardio-respiratory levels are reported for these athletes. Regardless of kickboxers' level, a high peak and mean anaerobic power output were reported. High-level kickboxing performance also requires well-developed muscle power in both the upper and lower limbs. Psychological factors contribute to success that requires high levels of self-confidence, motivation, dispositional hope and optimism, mental toughness/resiliency, and adaptive perfectionism. Psychological attributes also distinguished successful from less successful kickboxers. The activity-to-rest ratio was higher in elite (1:1) than both amateur and national-level (from 1:2 to 1:5) kickboxers, with no significant differences between rounds (round 1=1:4, and rounds 2 and 3=1:5) as well as between winners and losers in amateur and national-level simulated combats. These particular psychophysiological characteristics and performance aspects of kickboxers influence performance and could serve as guidance for training. Finally, kickboxing is characterized by chronic repetitive head trauma, which causes hypopituitarism due to traumatic brain injury (TBI). Future investigations into the physical, physiological and psychological

  15. Pharmacokinetic profiles of the active metamizole metabolites in healthy horses.

    PubMed

    Giorgi, M; Aupanun, S; Lee, H-K; Poapolathep, A; Rychshanova, R; Vullo, C; Faillace, V; Laus, F

    2017-04-01

    Metamizole (MT) is an analgesic and antipyretic drug labelled for use in humans, horses, cattle, swine and dogs. MT is rapidly hydrolysed to the active primary metabolite 4-methylaminoantipyrine (MAA). MAA is formed in much larger amounts compared with other minor metabolites. Among the other secondary metabolites, 4-aminoantipyrine (AA) is also relatively active. The aim of this research was to evaluate the pharmacokinetic profiles of MAA and AA after dose of 25 mg/kg MT by intravenous (i.v.) and intramuscular (i.m.) routes in healthy horses. Six horses were randomly allocated to two equally sized treatment groups according to a 2 × 2 crossover study design. Blood was collected at predetermined times within 24 h, and plasma was analysed by a validated HPLC-UV method. No behavioural changes or alterations in health parameters were observed in the i.v. or i.m. groups of animals during or after (up to 7 days) drug administration. Plasma concentrations of MAA after i.v. and i.m. administrations of MT were detectable from 5 min to 10 h in all the horses. Plasma concentrations of AA were detectable in the same range of time, but in smaller amounts. Maximum concentration (Cmax ), time to maximum concentration (Tmax ) and AUMC0-last of MAA were statistically different between the i.v. and i.m. groups. The AUCIM /AUCIV ratio of MAA was 1.06. In contrast, AUC0-last of AA was statistically different between the groups (P < 0.05) with an AUCIM /AUCIV ratio of 0.54. This study suggested that the differences in the MAA and AA plasma concentrations found after i.m. and i.v. administrations of MT might have minor consequences on the pharmacodynamics of the drug.

  16. Phytochemical profiles, antioxidant and antimicrobial activities of three Potentilla species

    PubMed Central

    2013-01-01

    Background Extracts from Potentilla species have been applied in traditional medicine and exhibit antioxidant, hypoglycemic, anti-inflammatory, antitumor and anti-ulcerogenic properties, but little has been known about the diversity of phytochemistry and pharmacology on this genus. This study investigated and compared the phytochemical profiles, antioxidant and antimicrobial activities of leaf extracts from three Potentilla species (Potentilla fruticosa, Potentilla glabra and Potentilla parvifolia) in order to discover new resources for lead structures and pharmaceutical products. Methods Chemical composition and content of six phenolic compounds were evaluated and determined by RP-HPLC; Total phenolic and total flavonoid content were determined using Folin-Ciocalteau colourimetric method and sodium borohydride/chloranil-based method (SBC); Antioxidant activities were determined using DPPH, ABTS and FRAP assays; Antimicrobial properties were investigated by agar dilution and mycelial growth rate method. Results The results showed hyperoside was the predominant phenolic compound in three Potentilla species by RP-HPLC assay, with the content of 8.86 (P. fruticosa), 2.56 (P. glabra) and 2.68 mg/g (P. parvifolia), respectively. The highest content of total identified phenolic compounds (hyperoside, (+)-catechin, caffeic acid, ferulic acid, rutin and ellagic acid) was observed in P. parvifolia (14.17 mg/g), follow by P. fruticosa (10.01 mg/g) and P. glabra (7.01 mg/g). P. fruticosa possessed the highest content of total phenolic (84.93 ± 0.50 mmol gallic acid equivalent/100 g) and total flavonoid (84.14 ± 0.03 mmol quercetin equivalent/100 g), which were in good correlation with its significant DPPHIC50 (16.87 μg/mL), ABTS (2763.48 μmol Trolox equivalent/g) and FRAP (1398.70 μmol Trolox equivalent/g) capacities. Furthermore, the effective methodology to distinguish the different species of Potentilla was also established by chromatographic fingerprint analysis for

  17. A time-dependent volume of distribution term used to describe linear concentration-time profiles.

    PubMed

    Colburn, W A

    1983-08-01

    The present study was conducted to develop, test, and apply a single exponential time-dependent volume of distribution function to describe the pharmacokinetics of compounds following instantaneous, zero-order and first order input. Simulations were used to show the applicability and flexibility of the equations. Experimental data from the literature were fitted using the equations developed in the present study. In addition, the results of these fitted curves were compared to the results of the original fitting procedures to compare and contrast the methods. In the present analysis, the change from an initial volume (V1) to the total volume (V T) is perceived as a simple exponential function. Therefore, a single exponential term to describe elimination and a single exponential term to describe the change from V1 to V T were used to describe the entire blood concentration profile. The results from present simulations and fitted data indicate that the transition from V1 to V T is a more continuous process than observed with classical methods and is consistent with results obtained from physiologic flow-limited models. This observation suggests that the present curve-fitting technique may be more akin to physiologic reality, in that it depicts the change from the initial volume of distribution to the total volume of distribution as a continuous exponential function which reflects the establishment of an equilibrium.

  18. Genotype-dependent variation of mitochondrial transcriptional profiles in interpopulation hybrids.

    PubMed

    Ellison, Christopher K; Burton, Ronald S

    2008-10-14

    Hybridization between populations can disrupt gene expression, frequently resulting in deleterious hybrid phenotypes. Reduced fitness in interpopulation hybrids of the marine copepod Tigriopus californicus has been traced to interactions between the nuclear and mitochondrial genomes. Here, we determine transcript levels of four to six genes involved in the mitochondrial oxidative phosphorylation pathway for a series of parental and inbred hybrid lines using RT-qPCR. Both nuclear and mitochondrial-encoded genes are included in the analysis. Although all genes studied are up-regulated under salinity stress, only expression of genes located on the mtDNA differed among lines. Because mitochondrial genes are transcribed by a dedicated RNA polymerase encoded in the nuclear genome, we compare transcript levels among hybrid lines with different combinations of mitochondrial RNA polymerase and mtDNA genotypes. Lines bearing certain mtDNA-mitochondrial RNA polymerase genotypic combinations show a diminished capacity to up-regulate mitochondrial genes in response to hypoosmotic stress. Effects on the transcriptional profile depend on the specific interpopulation cross and are correlated with viability effects. We hypothesize that disruption of the mitochondrial transcriptional system in F(2) hybrids may play a central role in hybrid breakdown.

  19. Genotype-dependent variation of mitochondrial transcriptional profiles in interpopulation hybrids

    PubMed Central

    Ellison, Christopher K.; Burton, Ronald S.

    2008-01-01

    Hybridization between populations can disrupt gene expression, frequently resulting in deleterious hybrid phenotypes. Reduced fitness in interpopulation hybrids of the marine copepod Tigriopus californicus has been traced to interactions between the nuclear and mitochondrial genomes. Here, we determine transcript levels of four to six genes involved in the mitochondrial oxidative phosphorylation pathway for a series of parental and inbred hybrid lines using RT-qPCR. Both nuclear and mitochondrial-encoded genes are included in the analysis. Although all genes studied are up-regulated under salinity stress, only expression of genes located on the mtDNA differed among lines. Because mitochondrial genes are transcribed by a dedicated RNA polymerase encoded in the nuclear genome, we compare transcript levels among hybrid lines with different combinations of mitochondrial RNA polymerase and mtDNA genotypes. Lines bearing certain mtDNA-mitochondrial RNA polymerase genotypic combinations show a diminished capacity to up-regulate mitochondrial genes in response to hypoosmotic stress. Effects on the transcriptional profile depend on the specific interpopulation cross and are correlated with viability effects. We hypothesize that disruption of the mitochondrial transcriptional system in F2 hybrids may play a central role in hybrid breakdown. PMID:18843106

  20. Distinct age and differentiation-state dependent metabolic profiles of oligodendrocytes under optimal and stress conditions

    PubMed Central

    Fuh, Shih-Chieh; Hossain, Shireen; Almazan, Guillermina; Multhaup, Gerhard; Healy, Luke M.; Kennedy, Timothy E.; Antel, Jack P.

    2017-01-01

    Within the microenvironment of multiple sclerosis lesions, oligodendrocytes are subject to metabolic stress reflecting effects of focal ischemia and inflammation. Previous studies have shown that under optimal conditions in vitro, the respiratory activity of human adult brain-derived oligodendrocytes is lower and more predominantly glycolytic compared to oligodendrocytes differentiated in vitro from post natal rat brain oligodendrocyte progenitor cells. In response to sub-lethal metabolic stress, adult human oligodendrocytes reduce overall energy production rate impacting the capacity to maintain myelination. Here, we directly compare the metabolic profiles of oligodendrocytes derived from adult rat brain with oligodendrocytes newly differentiated in vitro from oligodendrocyte progenitor cells obtained from the post natal rat brain, under both optimal culture and metabolic stress (low/no glucose) conditions. Oxygen consumption and extracellular acidification rates were measured using a Seahorse extracellular flux analyzer. Our findings indicate that under optimal conditions, adult rat oligodendrocytes preferentially use glycolysis whereas newly differentiated post natal rat oligodendrocytes, and the oligodendrocyte progenitor cells from which they are derived, mainly utilize oxidative phosphorylation to produce ATP. Metabolic stress increases the rate of ATP production via oxidative phosphorylation and significantly reduces glycolysis in adult oligodendrocytes. The rate of ATP production was relatively unchanged in newly differentiated post natal oligodendrocytes under these stress conditions, while it was significantly reduced in oligodendrocyte progenitor cells. Our study indicates that both age and maturation influence the metabolic profile under optimal and stressed conditions, emphasizing the need to consider these variables for in vitro studies that aim to model adult human disease. PMID:28792512

  1. Chromatin-dependent regulation of RNA polymerases II and III activity throughout the transcription cycle.

    PubMed

    Jordán-Pla, Antonio; Gupta, Ishaan; de Miguel-Jiménez, Lola; Steinmetz, Lars M; Chávez, Sebastián; Pelechano, Vicent; Pérez-Ortín, José E

    2015-01-01

    The particular behaviour of eukaryotic RNA polymerases along different gene regions and amongst distinct gene functional groups is not totally understood. To cast light onto the alternative active or backtracking states of RNA polymerase II, we have quantitatively mapped active RNA polymerases at a high resolution following a new biotin-based genomic run-on (BioGRO) technique. Compared with conventional profiling with chromatin immunoprecipitation, the analysis of the BioGRO profiles in Saccharomyces cerevisiae shows that RNA polymerase II has unique activity profiles at both gene ends, which are highly dependent on positioned nucleosomes. This is the first demonstration of the in vivo influence of positioned nucleosomes on transcription elongation. The particular features at the 5' end and around the polyadenylation site indicate that this polymerase undergoes extensive specific-activity regulation in the initial and final transcription elongation phases. The genes encoding for ribosomal proteins show distinctive features at both ends. BioGRO also provides the first nascentome analysis for RNA polymerase III, which indicates that transcription of tRNA genes is poorly regulated at the individual copy level. The present study provides a novel perspective of the transcription cycle that incorporates inactivation/reactivation as an important aspect of RNA polymerase dynamics.

  2. Comparative activity profiles of Thielavia terrestris and Trichoderma reesei cellulases

    SciTech Connect

    Tuse, D.; Mason, B.J.; Skinner, W.A.

    1980-10-01

    The successful utilization of cellulosic materials depends on the development of economically feasible processes for the literation of low molecular weight soluble products from the polymers. These soluble products, such as hexoses and pentoses, can then be utilized as substrates for the microbial or chemical product of fuels, food, and chemical feedstocks. In the enzymatic saccharification of cellulose, one of the major roadblocks to the development of commercially attractive processes has been the instability of the cellulase complex. It is desirable, for example, to operate the conversion systems at elevated temperatures, but environments with high thermal energy can significantly shorten enzyme half life. The authors have isolated a strain of the fungus Thielavia terrestris that possesses a complete cellulase system, and its enzymes were found to have remarkable thermal stability. The author presents a comparison of the activities of the T. terrestris enzymes with those of Trichoderma reesei.

  3. Activity-dependent plasticity of hippocampal place maps

    PubMed Central

    Schoenenberger, Philipp; O'Neill, Joseph; Csicsvari, Jozsef

    2016-01-01

    Hippocampal neurons encode a cognitive map of space. These maps are thought to be updated during learning and in response to changes in the environment through activity-dependent synaptic plasticity. Here we examine how changes in activity influence spatial coding in rats using halorhodopsin-mediated, spatially selective optogenetic silencing. Halorhoposin stimulation leads to light-induced suppression in many place cells and interneurons; some place cells increase their firing through disinhibition, whereas some show no effect. We find that place fields of the unaffected subpopulation remain stable. On the other hand, place fields of suppressed place cells were unstable, showing remapping across sessions before and after optogenetic inhibition. Disinhibited place cells had stable maps but sustained an elevated firing rate. These findings suggest that place representation in the hippocampus is constantly governed by activity-dependent processes, and that disinhibition may provide a mechanism for rate remapping. PMID:27282121

  4. On parameterization of spectral line profiles including the speed-dependence in the case of gas mixtures

    NASA Astrophysics Data System (ADS)

    Kochanov, V. P.

    2017-03-01

    The physically grounded parameterization of a line profile including the speed-dependence was performed. It was shown that two actual parameters of the quadratic speed-dependence appear in gas mixtures instead of a single parameter in a pure gas. One of the parameters is associated with hard elastic velocity-changing collisions; the second is connected with the other sorts of collisions. For comparable concentrations of gas species, they may differ by 50% and depend nonlinearly on partial gas pressures. The dimensionless line narrowing parameter also reveals nonlinear pressure-dependence. The computational expressions for the line profile including all main physical mechanisms of its forming in conditions of gas mixtures are derived.

  5. Proteomic profiling of metalloprotease activities with cocktails of active-site probes

    PubMed Central

    Sieber, Stephan A; Niessen, Sherry; Hoover, Heather S; Cravatt, Benjamin F

    2006-01-01

    Metalloproteases are a large, diverse class of enzymes involved in many physiological and disease processes. Metalloproteases are regulated by post-translational mechanisms that diminish the effectiveness of conventional genomic and proteomic methods for their functional characterization. Chemical probes directed at active sites offer a potential way to measure metalloprotease activities in biological systems; however, large variations in structure limit the scope of any single small-molecule probe aimed at profiling this enzyme class. Here, we address this problem by creating a library of metalloprotease-directed probes that show complementary target selectivity. These probes were applied as a ‘cocktail’ to proteomes and their labeling profiles were analyzed collectively using an advanced liquid chromatography-mass spectrometry platform. More than 20 metalloproteases were identified, including members from nearly all of the major branches of this enzyme class. These findings suggest that chemical proteomic methods can serve as a universal strategy to profile the activity of the metalloprotease superfamily in complex biological systems. PMID:16565715

  6. In vivo high-throughput profiling of CRISPR-Cpf1 activity.

    PubMed

    Kim, Hui K; Song, Myungjae; Lee, Jinu; Menon, A Vipin; Jung, Soobin; Kang, Young-Mook; Choi, Jae W; Woo, Euijeon; Koh, Hyun C; Nam, Jin-Wu; Kim, Hyongbum

    2017-02-01

    CRISPR from Prevotella and Francisella 1 (Cpf1) is an effector endonuclease of the class 2 CRISPR-Cas (clustered regularly interspaced short palindromic repeats-CRISPR-associated proteins) gene editing system. We developed a method for evaluating Cpf1 activity, based on target sequence composition in mammalian cells, in a high-throughput manner. A library of >11,000 target sequence and guide RNA pairs was delivered into human cells using lentiviral vectors. Subsequent delivery of Cpf1 into this cell library induced insertions and deletions (indels) at the integrated synthetic target sequences, which allowed en masse evaluation of Cpf1 activity by using deep sequencing. With this approach, we determined protospacer-adjacent motif sequences of two Cpf1 nucleases, one from Acidaminococcus sp. BV3L6 (hereafter referred to as AsCpf1) and the other from Lachnospiraceae bacterium ND2006 (hereafter referred to as LbCpf1). We also defined target-sequence-dependent activity profiles of AsCpf1, which enabled the development of a web tool that predicts the indel frequencies for given target sequences (http://big.hanyang.ac.kr/cindel). Both the Cpf1 characterization profile and the in vivo high-throughput evaluation method will greatly facilitate Cpf1-based genome editing.

  7. Arrhenius temperature dependence of in vitro tissue plasminogen activator thrombolysis

    NASA Astrophysics Data System (ADS)

    Shaw, George J.; Dhamija, Ashima; Bavani, Nazli; Wagner, Kenneth R.; Holland, Christy K.

    2007-06-01

    Stroke is a devastating disease and a leading cause of death and disability. Currently, the only FDA approved therapy for acute ischemic stroke is the intravenous administration of the thrombolytic medication, recombinant tissue plasminogen activator (tPA). However, this treatment has many contraindications and can have dangerous side effects such as intra-cerebral hemorrhage. These treatment limitations have led to much interest in potential adjunctive therapies, such as therapeutic hypothermia (T <= 35 °C) and ultrasound enhanced thrombolysis. Such interest may lead to combining these therapies with tPA to treat stroke, however little is known about the effects of temperature on the thrombolytic efficacy of tPA. In this work, we measure the temperature dependence of the fractional clot mass loss Δm(T) resulting from tPA exposure in an in vitro human clot model. We find that the temperature dependence is well described by an Arrhenius temperature dependence with an effective activation energy Eeff of 42.0 ± 0.9 kJ mole-1. Eeff approximates the activation energy of the plasminogen-to-plasmin reaction of 48.9 kJ mole-1. A model to explain this temperature dependence is proposed. These results will be useful in predicting the effects of temperature in future lytic therapies.

  8. Different signaling patterns contribute to loss of keratinocyte cohesion dependent on autoantibody profile in pemphigus.

    PubMed

    Walter, Elias; Vielmuth, Franziska; Rotkopf, Lukas; Sárdy, Miklós; Horváth, Orsolya N; Goebeler, Matthias; Schmidt, Enno; Eming, Rüdiger; Hertl, Michael; Spindler, Volker; Waschke, Jens

    2017-06-15

    Pemphigus is an autoimmune blistering skin disease caused primarily by autoantibodies against desmoglein (Dsg)1 and 3. Here, we characterized the mechanisms engaged by pemphigus IgG from patients with different clinical phenotypes and autoantibody profiles. All pemphigus vulgaris (PV) and pemphigus foliaceus (PF) IgG and AK23, a monoclonal mouse antibody against Dsg3, caused loss of cell cohesion, cytokeratin retraction and p38MAPK activation. Strong alterations in Dsg3 distribution were caused by mucosal (aDsg3 antibodies), mucocutaneous (aDsg1 + aDsg3) as well as atypical (aDsg3) PV-IgG. All PV-IgG fractions and AK23 compromised Dsg3 but not Dsg1 binding and enhanced Src activity. In contrast, rapid Ca(2+) influx and Erk activation were induced by mucocutaneous PV-IgG and pemphigus foliaceus (PF) IgG (aDsg1) whereas cAMP was increased by mucosal and mucocutaneous PV-IgG only. Selective inhibition of p38MAPK, Src or PKC blocked loss of keratinocyte cohesion in response to all autoantibody fractions whereas Erk inhibition was protective against mucocutaneous PV-IgG and PF-IgG only. These results demonstrate that signaling patterns parallel the clinical phenotype as some mechanisms involved in loss of cell cohesion are caused by antibodies targeting Dsg3 whereas others correlate with autoantibodies against Dsg1. The concept of key desmosome regulators may explain observations from several experimental models of pemphigus.

  9. The Increasing Impact of Activity-Based Protein Profiling in Plant Science.

    PubMed

    Morimoto, Kyoko; van der Hoorn, Renier A L

    2016-03-01

    The active proteome dictates plant physiology. Yet, active proteins are difficult to predict based on transcript or protein levels, because protein activities are regulated post-translationally in their microenvironments. Over the past 10 years, activity-based protein profiling (ABPP) is increasingly used in plant science. ABPP monitors the activities of hundreds of plant proteins using tagged chemical probes that react with the active site of proteins in a mechanism-dependent manner. Since labeling is covalent and irreversible, labeled proteins can be detected and identified on protein gels and by mass spectrometry using tagged fluorophores and/or biotin. Here, we discuss general concepts, approaches and practical considerations of ABPP, before we summarize the discoveries made using 40 validated probes representing 14 chemotypes that can monitor the active state of >4,500 plant proteins. These discoveries and new opportunities indicate that this emerging functional proteomic technology is a powerful discovery tool that will have an increasing impact on plant science. © The Author 2016. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oup.com.

  10. Effect of germination on phytochemical profiles and antioxidant activity of mung bean sprouts (Vigna radiata).

    PubMed

    Guo, Xinbo; Li, Tong; Tang, Kexuan; Liu, Rui Hai

    2012-11-07

    Epidemiological studies have shown that regular consumption of fruits and vegetables is associated with reduced risk of chronic diseases. It is recommended to increase consumption of fruits and vegetables to prevent chronic diseases related to free radical-induced oxidative stress. Different varieties of fruits and vegetables provide different vitamins, phenolics, flavonoids, minerals, and dietary fibers for optimal health benefits. Mung bean sprouts are one of the major vegetables in human diet. However, the profiles of phytochemicals and effect of germination on phytochemical content and antioxidant activity of mung bean sprouts have not been studied. The objective of this study was to determine the effect of germination on phytochemical profiles and antioxidant activity of mung bean sprouts. Germination of mung beans dramatically increased vitamin C content in mung bean sprouts in a time-dependent manner and reached the peak on day 8 of germination up to 285 mg/100 g DW, almost 24 times higher than the initial concentration in mung bean seeds (p < 0.05). On fresh weight basis, one serving of mung bean sprouts (about 104 g) provides 21.6 mg of vitamin C, which could meet 36% of Daily Value (DV). In addition, the germination dramatically increased total phenolic compounds and total flavonoids in mung bean sprouts in a time-dependent manner, up to 4.5 and 6.8 times higher than the original concentration of mung bean seeds, respectively. Quercetin-3-O-glucoside content was significantly increased in mung bean sprouts after germination. The total antioxidant activity of mung bean sprouts was increased by 6 times higher than that of mung bean seeds. Therefore, the germination of mung bean sprouts significantly increased phytochemical content, vitamin C content, and antioxidant activity.

  11. Arrestin-Dependent Activation of JNK Family Kinases

    PubMed Central

    Zhan, Xuanzhi; Kook, Seunghyi; Gurevich, Eugenia V.

    2015-01-01

    The activity of all mitogen-activated protein kinases (MAPKs) is stimulated via phosphorylation by upstream MAPK kinases (MAPKK), which are in their turn activated via phosphorylation by MAPKK kinases (MAPKKKs). The cells ensure the specificity of signaling in these cascades by employing a variety of scaffolding proteins that bind matching MAPKKKs, MAPKKs, and MAPKs. All four vertebrate arrestin subtypes bind JNK3, but only arrestin-3 serves as a scaffold, promoting JNK3 activation in intact cells. Arrestin-3-mediated JNK3 activation does not depend on arrestin-3 interaction with G protein-coupled receptors (GPCRs), as demonstrated by the ability of some arrestin mutants that cannot bind receptors to activate JNK3, whereas certain mutants with enhanced GPCR binding fail to promote JNK3 activation. Recent findings suggest that arrestin-3 directly binds both MAPKKs necessary for JNK activation and facilitates JNK3 phosphorylation at both Thr (by MKK4) and Tyr (by MKK7). JNK3 is expressed in a limited set of cell types, whereas JNK1 and JNK2 isoforms are as ubiquitous as arrestin-3. Recent study showed that arrestin-3 facilitates the activation of JNK1 and JNK2, scaffolding MKK4/7-JNK1/2/3 signaling complexes. In all cases, arrestin-3 acts by bringing the kinases together: JNK phosphorylation shows biphasic dependence on arrestin-3, being enhanced at lower and suppressed at supraoptimal concentrations. Thus, arrestin-3 regulates the activity of multiple JNK isoforms, suggesting that it might play a role in survival and apoptosis of all cell types. PMID:24292834

  12. Comparative assessment of performance and genome dependence among phylogenetic profiling methods

    PubMed Central

    Snitkin, Evan S; Gustafson, Adam M; Mellor, Joseph; Wu, Jie; DeLisi, Charles

    2006-01-01

    Background The rapidly increasing speed with which genome sequence data can be generated will be accompanied by an exponential increase in the number of sequenced eukaryotes. With the increasing number of sequenced eukaryotic genomes comes a need for bioinformatic techniques to aid in functional annotation. Ideally, genome context based techniques such as proximity, fusion, and phylogenetic profiling, which have been so successful in prokaryotes, could be utilized in eukaryotes. Here we explore the application of phylogenetic profiling, a method that exploits the evolutionary co-occurrence of genes in the assignment of functional linkages, to eukaryotic genomes. Results In order to evaluate the performance of phylogenetic profiling in eukaryotes, we assessed the relative performance of commonly used profile construction techniques and genome compositions in predicting functional linkages in both prokaryotic and eukaryotic organisms. When predicting linkages in E. coli with a prokaryotic profile, the use of continuous values constructed from transformed BLAST bit-scores performed better than profiles composed of discretized E-values; the use of discretized E-values resulted in more accurate linkages when using S. cerevisiae as the query organism. Extending this analysis by incorporating several eukaryotic genomes in profiles containing a majority of prokaryotes resulted in similar overall accuracy, but with a surprising reduction in pathway diversity among the most significant linkages. Furthermore, the application of phylogenetic profiling using profiles composed of only eukaryotes resulted in the loss of the strong correlation between common KEGG pathway membership and profile similarity score. Profile construction methods, orthology definitions, ontology and domain complexity were explored as possible sources of the poor performance of eukaryotic profiles, but with no improvement in results. Conclusion Given the current set of completely sequenced eukaryotic

  13. Phenolic and carotenoid profiles and antiproliferative activity of foxtail millet.

    PubMed

    Zhang, Li Zhen; Liu, Rui Hai

    2015-05-01

    Commonly consumed foxtail millet varieties Jingu28 and Jingu34 were compared in terms of phytochemical composition, antioxidant property, and antiproliferative activity. The cellular antioxidant activity (CAA) was evaluated based on HepG2 cell cultivation. Antiproliferative properties against HepG2 and MDA cell were assayed by methylene blue assay. Total phenolic content (TPC) was 78.79 and 114.22 mg gallic acid equiv/100 g DW in Jingu28 and Jingu34. Both varieties contained ferulic acid, chlorogenic acid, caffeic acid and p-coumaric acid, syringic acid. Xanthophylls and zeaxanthin were also detected. Peroxyl radical scavenging capacity of the foxtail millet were 228.13 (Jingu28) and 355.03 (Jingu34) μmol of vitamin C equiv/100 g, respectively. CAA values of the foxtail millet varieties ranged from 1.52 to 8.97 μmol quercetin equiv/100 g DW. The proliferation of MDA and HepG2 cancer cells were significantly inhibited in a dose-dependent manner after exposure to Jingu28 and Jingu34 extractions.

  14. PHARMACOLOGICAL SIRT1 ACTIVATION IMPROVES MORTALITY AND MARKEDLY ALTERS TRANSCRIPTIONAL PROFILES THAT ACCOMPANY EXPERIMENTAL SEPSIS.

    PubMed

    Opal, Steven M; Ellis, James L; Suri, Vipin; Freudenberg, Johannes M; Vlasuk, George P; Li, Yong; Chahin, Abdullah B; Palardy, John E; Parejo, Nicholas; Yamamoto, Michelle; Chahin, Abdulrahman; Kessimian, Noubar

    2016-04-01

    The sirtuin family consists of seven NAD+-dependent enzymes affecting a broad array of regulatory protein networks by primarily catalyzing the deacetylation of key lysine residues in regulatory proteins. The enzymatic activity of SIRT1 can be enhanced by small molecule activators known as SIRT1 activator compounds (STACs). We tested the therapeutic potential of the STAC SRT3025 in two preclinical models of severe infection, the murine cecal ligation and puncture (CLP) model to induce peritonitis and intratracheal installation of Streptococcus pneumoniae to induce severe bacterial pneumonia. SRT3025 provided significant survival benefits over vehicle control in both the peritonitis and pneumococcal pneumonia models when administered with appropriate antimicrobial agents. The survival benefit of SRT3025 in the CLP model was absent in SIRT1 knockout showing the SIRT1 dependency of SRT3025's effects. SRT3025 administration promoted bacterial clearance and significantly reduced inflammatory cytokines from the lungs of animals challenged with S. pneumoniae. SRT3025 treatment was also accompanied by striking changes in the transcription profiles in multiple inflammatory and metabolic pathways in liver, spleen, small bowel, and lung tissue. Remarkably, these organ-specific changes in the transcriptome analyses were similar following CLP or pneumococcal challenge despite different sets of pathogens at disparate sites of infection. Pharmacologic activation of SIRT1 modulates the innate host response and could represent a novel treatment strategy for severe infection.

  15. Sirt1 Activation Markedly Alters Transcription Profiles and Improves Outcome in Experimental Sepsis.

    PubMed

    Opal, Steven; Ellis, James L; Suri, Vipin; Freudenberg, Johannes M; Vlasuk, George P; Li, Yong; Chahin, Abdullah B; Palardy, John E; Parejo, Nicholas; Yamamoto, Michelle; Chahin, Abdulrahman; Kessimian, Noubar

    2015-11-16

    The sirtuin family consists of seven NAD+-dependent enzymes affecting a broad array of regulatory protein networks by primarily catalyzing the deacetylation of key lysine residues in regulatory proteins. The enzymatic activity of SIRT1 can be enhanced by small molecule activators known as SIRT1 activator compounds (STACs). We tested the therapeutic potential of the STAC SRT3025 in two preclinical models of severe infection, the murine cecal ligation and puncture (CLP) model to induce peritonitis and intra-tracheal installation of Streptococcus pneumoniae to induce severe bacterial pneumonia. SRT3025 provided significant survival benefits over vehicle control in both the peritonitis and pneumococcal pneumonia models when administered with appropriate antimicrobial agents. The survival benefit of SRT3025 in the CLP model was absent in SIRT1 knock-out showing the SIRT1-dependency of SRT3025's effects. SRT3025 administration promoted bacterial clearance and significantly reduced inflammatory cytokines from the lungs of animals challenged with S. pneumoniae. SRT3025 treatment was also accompanied by striking changes in the transcription profiles in multiple inflammatory and metabolic pathways in liver, spleen, small bowel and lung tissue. Remarkably, these organ-specific changes in the transcriptome analyses were similar following CLP or pneumococcal challenge despite different sets of pathogens at disparate sites of infection. Pharmacologic activation of SIRT1 modulates the innate host response and could represent a novel treatment strategy for severe infection.

  16. Proteomic profile of Aspergillus flavus in response to water activity.

    PubMed

    Zhang, Feng; Zhong, Hong; Han, Xiaoyun; Guo, Zhenni; Yang, Weiqiang; Liu, Yongfeng; Yang, Kunlong; Zhuang, Zhenhong; Wang, Shihua

    2015-03-01

    Aspergillus flavus, a common contaminant of crops and stored grains, can produce aflatoxins that are harmful to humans and other animals. Water activity (aw) is one of the key factors influencing both fungal growth and mycotoxin production. In this study, we used the isobaric tagging for relative and absolute quantitation (iTRAQ) technique to investigate the effect of aw on the proteomic profile of A. flavus. A total of 3566 proteins were identified, of which 837 were differentially expressed in response to variations in aw. Among these 837 proteins, 403 were over-expressed at 0.99 aw, whereas 434 proteins were over-expressed at 0.93 aw. According to Gene Ontology (GO) analysis, the secretion of extracellular hydrolases increased as aw was raised, suggesting that extracellular hydrolases may play a critical role in induction of aflatoxin biosynthesis. On the basis of Clusters of Orthologous Groups (COG) and Kyoto Encyclopedia of Genes and Genomes (KEGG) categorizations, we identified an exportin protein, KapK, that may down-regulate aflatoxin biosynthesis by changing the location of NirA. Finally, we considered the role of two osmotic stress-related proteins (Sln1 and Glo1) in the Hog1 pathway and investigated the expression patterns of proteins related to aflatoxin biosynthesis. The data uncovered in this study are critical for understanding the effect of water stress on toxin production and for the development of strategies to control toxin contamination of agricultural products. Copyright © 2014 The British Mycological Society. Published by Elsevier Ltd. All rights reserved.

  17. The Effects of Physical Training on Blood Lipid Profiles in Adolescents with Insulin-Dependent Diabetes Mellitus.

    ERIC Educational Resources Information Center

    Campaigne, B. N.; And Others

    1985-01-01

    Fourteen adolescents with insulin-dependent diabetes mellitus (IDDM) participated in a 12-week exercise program to determine whether such training would bring about changes in blood lipid and lipoprotein profiles. The findings support the beneficial effects of regular exercise for individuals with IDDM. (MT)

  18. The Effects of Physical Training on Blood Lipid Profiles in Adolescents with Insulin-Dependent Diabetes Mellitus.

    ERIC Educational Resources Information Center

    Campaigne, B. N.; And Others

    1985-01-01

    Fourteen adolescents with insulin-dependent diabetes mellitus (IDDM) participated in a 12-week exercise program to determine whether such training would bring about changes in blood lipid and lipoprotein profiles. The findings support the beneficial effects of regular exercise for individuals with IDDM. (MT)

  19. Solar activity dependence of nightside aurora in winter conditions

    NASA Astrophysics Data System (ADS)

    Zhou, Su; Luan, Xiaoli; Dou, Xiankang

    2016-02-01

    The dependence of the nightside (21:00-03:00 MLT; magnetic local time) auroral energy flux on solar activity was quantitatively studied for winter/dark and geomagnetically quiet conditions. Using data combined from Thermosphere, Ionosphere, Mesosphere Energetics and Dynamics/Global Ultraviolet Imager and Defense Meteorological Satellite Program/Special Sensor Ultraviolet Spectrographic Imager observations, we separated the effects of geomagnetic activity from those of solar flux on the nightside auroral precipitation. The results showed that the nightside auroral power was reduced by ~42% in solar maximum (F10.7 = 200 sfu; solar flux unit 1 sfu = 10-22 W m-2 Hz-1) with respect to that under solar minimum (F10.7 = 70 sfu) for the Kp = 1 condition, and this change rate became less (~21%) for the Kp = 3 condition. In addition, the solar cycle dependence of nightside auroral power was similar with that from both the premidnight (21:00-23:00 MLT) and postmidnight (01:00-03:00 MLT) sectors. These results indicated that as the ionospheric ionization increases with the enhanced auroral and geomagnetic activities, the solar activity dependences of nightside auroral power become weaker, at least under geomagnetically quiet conditions.

  20. Cyclic AMP-dependent protein kinase activity in Trypanosoma cruzi.

    PubMed Central

    Ulloa, R M; Mesri, E; Esteva, M; Torres, H N; Téllez-Iñón, M T

    1988-01-01

    A cyclic AMP-dependent protein kinase activity from epimastigote forms of Trypanosoma cruzi was characterized. Cytosolic extracts were chromatographed on DEAE-cellulose columns, giving two peaks of kinase activity, which were eluted at 0.15 M- and 0.32 M-NaCl respectively. The second activity peak was stimulated by nanomolar concentrations of cyclic AMP. In addition, a cyclic AMP-binding protein co-eluted with the second kinase activity peak. Cyclic AMP-dependent protein kinase activity was further purified by gel filtration, affinity chromatography on histone-agarose and cyclic AMP-agarose, as well as by chromatography on CM-Sephadex. The enzyme ('holoenzyme') could be partially dissociated into two different components: 'catalytic' and 'regulatory'. The 'regulatory' component had specific binding for cyclic AMP, and it inhibited phosphotransferase activity of the homologous 'catalytic component' or of the 'catalytic subunit' from bovine heart. Cyclic AMP reversed these inhibitions. A 'holoenzyme preparation' was phosphorylated in the absence of exogenous phosphate acceptor and analysed by polyacrylamide-gel electrophoresis. A 56 kDa band was phosphorylated. The same preparation was analysed by Western blotting, by using polyclonal antibodies to the regulatory subunits of protein kinases type I or II. Both antibodies reacted with the 56 kDa band. Images Fig. 7. Fig. 8. PMID:2848508

  1. A network model for activity-dependent sleep regulation.

    PubMed

    Roy, Sandip; Krueger, James M; Rector, David M; Wan, Yan

    2008-08-07

    We develop and characterize a dynamical network model for activity-dependent sleep regulation. Specifically, in accordance with the activity-dependent theory for sleep, we view organism sleep as emerging from the local sleep states of functional units known as cortical columns; these local sleep states evolve through integration of local activity inputs, loose couplings with neighboring cortical columns, and global regulation (e.g. by the circadian clock). We model these cortical columns as coupled or networked activity-integrators that transition between sleep and waking states based on thresholds on the total activity. The model dynamics for three canonical experiments (which we have studied both through simulation and system-theoretic analysis) match with experimentally observed characteristics of the cortical-column network. Most notably, assuming connectedness of the network graph, our model predicts the recovery of the columns to a synchronized state upon temporary overstimulation of a single column and/or randomization of the initial sleep and activity-integration states. In analogy with other models for networked oscillators, our model also predicts the possibility for such phenomena as mode-locking.

  2. Network-dependent modulation of brain activity during sleep.

    PubMed

    Watanabe, Takamitsu; Kan, Shigeyuki; Koike, Takahiko; Misaki, Masaya; Konishi, Seiki; Miyauchi, Satoru; Miyahsita, Yasushi; Masuda, Naoki

    2014-09-01

    Brain activity dynamically changes even during sleep. A line of neuroimaging studies has reported changes in functional connectivity and regional activity across different sleep stages such as slow-wave sleep (SWS) and rapid-eye-movement (REM) sleep. However, it remains unclear whether and how the large-scale network activity of human brains changes within a given sleep stage. Here, we investigated modulation of network activity within sleep stages by applying the pairwise maximum entropy model to brain activity obtained by functional magnetic resonance imaging from sleeping healthy subjects. We found that the brain activity of individual brain regions and functional interactions between pairs of regions significantly increased in the default-mode network during SWS and decreased during REM sleep. In contrast, the network activity of the fronto-parietal and sensory-motor networks showed the opposite pattern. Furthermore, in the three networks, the amount of the activity changes throughout REM sleep was negatively correlated with that throughout SWS. The present findings suggest that the brain activity is dynamically modulated even in a sleep stage and that the pattern of modulation depends on the type of the large-scale brain networks.

  3. Activity-Dependent Human Brain Coding/Noncoding Gene Regulatory Networks

    PubMed Central

    Lipovich, Leonard; Dachet, Fabien; Cai, Juan; Bagla, Shruti; Balan, Karina; Jia, Hui; Loeb, Jeffrey A.

    2012-01-01

    While most gene transcription yields RNA transcripts that code for proteins, a sizable proportion of the genome generates RNA transcripts that do not code for proteins, but may have important regulatory functions. The brain-derived neurotrophic factor (BDNF) gene, a key regulator of neuronal activity, is overlapped by a primate-specific, antisense long noncoding RNA (lncRNA) called BDNFOS. We demonstrate reciprocal patterns of BDNF and BDNFOS transcription in highly active regions of human neocortex removed as a treatment for intractable seizures. A genome-wide analysis of activity-dependent coding and noncoding human transcription using a custom lncRNA microarray identified 1288 differentially expressed lncRNAs, of which 26 had expression profiles that matched activity-dependent coding genes and an additional 8 were adjacent to or overlapping with differentially expressed protein-coding genes. The functions of most of these protein-coding partner genes, such as ARC, include long-term potentiation, synaptic activity, and memory. The nuclear lncRNAs NEAT1, MALAT1, and RPPH1, composing an RNAse P-dependent lncRNA-maturation pathway, were also upregulated. As a means to replicate human neuronal activity, repeated depolarization of SY5Y cells resulted in sustained CREB activation and produced an inverse pattern of BDNF-BDNFOS co-expression that was not achieved with a single depolarization. RNAi-mediated knockdown of BDNFOS in human SY5Y cells increased BDNF expression, suggesting that BDNFOS directly downregulates BDNF. Temporal expression patterns of other lncRNA-messenger RNA pairs validated the effect of chronic neuronal activity on the transcriptome and implied various lncRNA regulatory mechanisms. lncRNAs, some of which are unique to primates, thus appear to have potentially important regulatory roles in activity-dependent human brain plasticity. PMID:22960213

  4. Temperature dependence of oxygen evolution through catalase-like activity of horseradish peroxidase

    NASA Astrophysics Data System (ADS)

    Popović-Bijelić, A.; Bijelić, G.; Kolar-Anić, Lj.; Vukojević, V.

    2007-09-01

    By experimental investigations of the temperature dependence of catalase-like activity of horseradish peroxidase in the temperature range 278 328 K, different kinetic profiles for oxygen evolution were found below and above 298 K. Extension of the model is proposed to account for these observations. By numeric simulations of the reaction kinetics at different temperatures, it was found that enhanced evaporation of molecular oxygen from the reaction solution is the main root through which oxygen is lost at elevated temperatures in laboratory conditions.

  5. Stress versus temperature dependence of activation energies for creep

    NASA Technical Reports Server (NTRS)

    Freed, A. D.; Raj, S. V.; Walker, K. P.

    1992-01-01

    The activation energy for creep at low stresses and elevated temperatures is associated with lattice diffusion, where the rate controlling mechanism for deformation is dislocation climb. At higher stresses and intermediate temperatures, the rate controlling mechanism changes from dislocation climb to obstacle-controlled dislocation glide. Along with this change in deformation mechanism occurs a change in the activation energy. When the rate controlling mechanism for deformation is obstacle-controlled dislocation glide, it is shown that a temperature-dependent Gibbs free energy does better than a stress-dependent Gibbs free energy in correlating steady-state creep data for both copper and LiF-22mol percent CaF2 hypereutectic salt.

  6. Stress versus temperature dependent activation energies in creep

    NASA Technical Reports Server (NTRS)

    Freed, A. D.; Raj, S. V.; Walker, K. P.

    1990-01-01

    The activation energy for creep at low stresses and elevated temperatures is lattice diffusion, where the rate controlling mechanism for deformation is dislocation climb. At higher stresses and intermediate temperatures, the rate controlling mechanism changes from that of dislocation climb to one of obstacle-controlled dislocation glide. Along with this change, there occurs a change in the activation energy. It is shown that a temperature-dependent Gibbs free energy does a good job of correlating steady-state creep data, while a stress-dependent Gibbs free energy does a less desirable job of correlating the same data. Applications are made to copper and a LiF-22 mol. percent CaF2 hypereutectic salt.

  7. ‘Dopamine-first’ mechanism enables the rational engineering of the norcoclaurine synthase aldehyde activity profile

    PubMed Central

    Lichman, Benjamin R; Gershater, Markus C; Lamming, Eleanor D; Pesnot, Thomas; Sula, Altin; Keep, Nicholas H; Hailes, Helen C; Ward, John M

    2015-01-01

    Norcoclaurine synthase (NCS) (EC 4.2.1.78) catalyzes the Pictet–Spengler condensation of dopamine and an aldehyde, forming a substituted (S)-tetrahydroisoquinoline, a pharmaceutically important moiety. This unique activity has led to NCS being used for both in vitro biocatalysis and in vivo recombinant metabolism. Future engineering of NCS activity to enable the synthesis of diverse tetrahydroisoquinolines is dependent on an understanding of the NCS mechanism and kinetics. We assess two proposed mechanisms for NCS activity: (a) one based on the holo X-ray crystal structure and (b) the ‘dopamine-first’ mechanism based on computational docking. Thalictrum flavum NCS variant activities support the dopamine-first mechanism. Suppression of the non-enzymatic background reaction reveals novel kinetic parameters for NCS, showing it to act with low catalytic efficiency. This kinetic behaviour can account for the ineffectiveness of recombinant NCS in in vivo systems, and also suggests NCS may have an in planta role as a metabolic gatekeeper. The amino acid substitution L76A, situated in the proposed aldehyde binding site, results in the alteration of the enzyme's aldehyde activity profile. This both verifies the dopamine-first mechanism and demonstrates the potential for the rational engineering of NCS activity. PMID:25620686

  8. Online Activity Levels Are Related to Caffeine Dependency.

    PubMed

    Phillips, James G; Landhuis, C Erik; Shepherd, Daniel; Ogeil, Rowan P

    2016-05-01

    Online activity could serve in the future as behavioral markers of emotional states for computer systems (i.e., affective computing). Hence, this study considered relationships between self-reported stimulant use and online study patterns. Sixty-two undergraduate psychology students estimated their daily caffeine use, and this was related to study patterns as tracked by their use of a Learning Management System (Blackboard). Caffeine dependency was associated with less time spent online, lower rates of file access, and fewer online activities completed. Reduced breadth or depth of processing during work/study could be used as a behavioral marker of stimulant use.

  9. The Zinc-Dependent Protease Activity of the Botulinum Neurotoxins

    PubMed Central

    Lebeda, Frank J.; Cer, Regina Z.; Mudunuri, Uma; Stephens, Robert; Singh, Bal Ram; Adler, Michael

    2010-01-01

    The botulinum neurotoxins (BoNT, serotypes A-G) are some of the most toxic proteins known and are the causative agents of botulism. Following exposure, the neurotoxin binds and enters peripheral cholinergic nerve endings and specifically and selectively cleaves one or more SNARE proteins to produce flaccid paralysis. This review centers on the kinetics of the Zn-dependent proteolytic activities of these neurotoxins, and briefly describes inhibitors, activators and factors underlying persistence of toxin action. Some of the structural, enzymatic and inhibitor data that are discussed here are available at the botulinum neurotoxin resource, BotDB (http://botdb.abcc.ncifcrf.gov). PMID:22069621

  10. Peptide-size dependent active transport in the proteasome

    NASA Astrophysics Data System (ADS)

    Zaikin, A.; Pöschel, T.

    2005-03-01

    We investigate the transport of proteins inside the proteasome and propose an active-transport mechanism based on a spatially asymmetric interaction potential of peptide chains. The transport is driven by fluctuations which are always present in such systems. We compute the peptide-size dependent transport rate which is essential for the functioning of the proteasome. In agreement with recent experiments, varying temperature changes the transport mechanism qualitatively.

  11. Allosteric activation of apicomplexan calcium-dependent protein kinases

    SciTech Connect

    Ingram, Jessica R.; Knockenhauer, Kevin E.; Markus, Benedikt M.; Mandelbaum, Joseph; Ramek, Alexander; Shan, Yibing; Shaw, David E.; Schwartz, Thomas U.; Ploegh, Hidde L.; Lourido, Sebastian

    2015-08-24

    Calcium-dependent protein kinases (CDPKs) comprise the major group of Ca2+-regulated kinases in plants and protists. It has long been assumed that CDPKs are activated, like other Ca2+-regulated kinases, by derepression of the kinase domain (KD). However, we found that removal of the autoinhibitory domain from Toxoplasma gondii CDPK1 is not sufficient for kinase activation. From a library of heavy chain-only antibody fragments (VHHs), we isolated an antibody (1B7) that binds TgCDPK1 in a conformation-dependent manner and potently inhibits it. We uncovered the molecular basis for this inhibition by solving the crystal structure of the complex and simulating, through molecular dynamics, the effects of 1B7–kinase interactions. In contrast to other Ca2+-regulated kinases, the regulatory domain of TgCDPK1 plays a dual role, inhibiting or activating the kinase in response to changes in Ca2+ concentrations. We propose that the regulatory domain of TgCDPK1 acts as a molecular splint to stabilize the otherwise inactive KD. This dependence on allosteric stabilization reveals a novel susceptibility in this important class of parasite enzymes.

  12. Allosteric activation of apicomplexan calcium-dependent protein kinases

    DOE PAGES

    Ingram, Jessica R.; Knockenhauer, Kevin E.; Markus, Benedikt M.; ...

    2015-08-24

    Calcium-dependent protein kinases (CDPKs) comprise the major group of Ca2+-regulated kinases in plants and protists. It has long been assumed that CDPKs are activated, like other Ca2+-regulated kinases, by derepression of the kinase domain (KD). However, we found that removal of the autoinhibitory domain from Toxoplasma gondii CDPK1 is not sufficient for kinase activation. From a library of heavy chain-only antibody fragments (VHHs), we isolated an antibody (1B7) that binds TgCDPK1 in a conformation-dependent manner and potently inhibits it. We uncovered the molecular basis for this inhibition by solving the crystal structure of the complex and simulating, through molecular dynamics,more » the effects of 1B7–kinase interactions. In contrast to other Ca2+-regulated kinases, the regulatory domain of TgCDPK1 plays a dual role, inhibiting or activating the kinase in response to changes in Ca2+ concentrations. We propose that the regulatory domain of TgCDPK1 acts as a molecular splint to stabilize the otherwise inactive KD. This dependence on allosteric stabilization reveals a novel susceptibility in this important class of parasite enzymes.« less

  13. Allosteric activation of apicomplexan calcium-dependent protein kinases

    PubMed Central

    Ingram, Jessica R.; Knockenhauer, Kevin E.; Markus, Benedikt M.; Mandelbaum, Joseph; Ramek, Alexander; Shan, Yibing; Shaw, David E.; Schwartz, Thomas U.; Ploegh, Hidde L.; Lourido, Sebastian

    2015-01-01

    Calcium-dependent protein kinases (CDPKs) comprise the major group of Ca2+-regulated kinases in plants and protists. It has long been assumed that CDPKs are activated, like other Ca2+-regulated kinases, by derepression of the kinase domain (KD). However, we found that removal of the autoinhibitory domain from Toxoplasma gondii CDPK1 is not sufficient for kinase activation. From a library of heavy chain-only antibody fragments (VHHs), we isolated an antibody (1B7) that binds TgCDPK1 in a conformation-dependent manner and potently inhibits it. We uncovered the molecular basis for this inhibition by solving the crystal structure of the complex and simulating, through molecular dynamics, the effects of 1B7–kinase interactions. In contrast to other Ca2+-regulated kinases, the regulatory domain of TgCDPK1 plays a dual role, inhibiting or activating the kinase in response to changes in Ca2+ concentrations. We propose that the regulatory domain of TgCDPK1 acts as a molecular splint to stabilize the otherwise inactive KD. This dependence on allosteric stabilization reveals a novel susceptibility in this important class of parasite enzymes. PMID:26305940

  14. Pyrethroid activity-based probes for profiling cytochrome P450 activities associated with insecticide interactions

    PubMed Central

    Ismail, Hanafy M.; O’Neill, Paul M.; Hong, David W.; Finn, Robert D.; Henderson, Colin J.; Wright, Aaron T.; Cravatt, Benjamin F.; Hemingway, Janet; Paine, Mark J. I.

    2013-01-01

    Pyrethroid insecticides are used to control diseases spread by arthropods. We have developed a suite of pyrethroid mimetic activity-based probes (PyABPs) to selectively label and identify P450s associated with pyrethroid metabolism. The probes were screened against pyrethroid-metabolizing and nonmetabolizing mosquito P450s, as well as rodent microsomes, to measure labeling specificity, plus cytochrome P450 oxidoreductase and b5 knockout mouse livers to validate P450 activation and establish the role for b5 in probe activation. Using PyABPs, we were able to profile active enzymes in rat liver microsomes and identify pyrethroid-metabolizing enzymes in the target tissue. These included P450s as well as related detoxification enzymes, notably UDP-glucuronosyltransferases, suggesting a network of associated pyrethroid-metabolizing enzymes, or “pyrethrome.” Considering the central role P450s play in metabolizing insecticides, we anticipate that PyABPs will aid in the identification and profiling of P450s associated with insecticide pharmacology in a wide range of species, improving understanding of P450–insecticide interactions and aiding the development of unique tools for disease control. PMID:24248381

  15. Pyrethroid Activity-Based Probes for Profiling Cytochrome P450 Activities Associated with Insecticide Interactions

    SciTech Connect

    Ismail, Hanafy M.; O'Neill, Paul M.; Hong, David; Finn, Robert; Henderson, Colin; Wright, Aaron T.; Cravatt, Benjamin; Hemingway, Janet; Paine, Mark J.

    2014-01-18

    Pyrethroid insecticides are used to control a diverse spectrum of diseases spread by arthropods. We have developed a suite of pyrethroid mimetic activity based probes (PyABPs) to selectively label and identify P450s associated with pyrethroid metabolism. The probes were screened against pyrethroid metabolizing and non-metabolizing mosquito P450s, as well as rodent microsomes to measure labeling specificity, plus CPR and b5 knockout mouse livers to validate P450 activation and establish the role for b5 in probe activation. Using a deltamethrin mimetic PyABP we were able to profile active enzymes in rat liver microsomes and identify pyrethroid metabolizing enzymes in the target tissue. The most reactive enzyme was a P450, CYP2C11, which is known to metabolize deltamethrin. Furthermore, several other pyrethroid metabolizers were identified (CYPs 2C6, 3A4, 2C13 and 2D1) along with related detoxification enzymes, notably UDP-g’s 2B1 - 5, suggesting a network of associated pyrethroid metabolizing enzymes, or ‘pyrethrome’. Considering the central role that P450s play in metabolizing insecticides, we anticipate that PyABPs will aid the identification and profiling of P450s associated with insecticide pharmacology in a wide range of species, improving understanding of P450-insecticide interactions and aiding the development of new tools for disease control.

  16. Quantitative site-specific ADP-ribosylation profiling of DNA-dependent PARPs.

    PubMed

    Gagné, Jean-Philippe; Ethier, Chantal; Defoy, Daniel; Bourassa, Sylvie; Langelier, Marie-France; Riccio, Amanda A; Pascal, John M; Moon, Kyung-Mee; Foster, Leonard J; Ning, Zhibin; Figeys, Daniel; Droit, Arnaud; Poirier, Guy G

    2015-06-01

    An important feature of poly(ADP-ribose) polymerases (PARPs) is their ability to readily undergo automodification upon activation. Although a growing number of substrates were found to be poly(ADP-ribosyl)ated, including histones and several DNA damage response factors, PARPs themselves are still considered as the main acceptors of poly(ADP-ribose). By monitoring spectral counts of specific hydroxamic acid signatures generated after the conversion of the ADP-ribose modification onto peptides by hydroxylamine hydrolysis, we undertook a thorough mass spectrometry mapping of the glutamate and aspartate ADP-ribosylation sites onto automodified PARP-1, PARP-2 and PARP-3. Thousands of hydroxamic acid-conjugated peptides were identified with high confidence and ranked based on their spectral count. This semi-quantitative approach allowed us to locate the preferentially targeted residues in DNA-dependent PARPs. In contrast to what has been reported in the literature, automodification of PARP-1 is not predominantly targeted towards its BRCT domain. Our results show that interdomain linker regions that connect the BRCT to the WGR module and the WGR to the PRD domain undergo prominent ADP-ribosylation during PARP-1 automodification. We also found that PARP-1 efficiently automodifies the D-loop structure within its own catalytic fold. Interestingly, additional major ADP-ribosylation sites were identified in functional domains of PARP-1, including all three zinc fingers. Similar to PARP-1, specific residues located within the catalytic sites of PARP-2 and PARP-3 are major targets of automodification following their DNA-dependent activation. Together our results suggest that poly(ADP-ribosyl)ation hot spots make a dominant contribution to the overall automodification process.

  17. Dose-dependent effects of caffeine in human Sertoli cells metabolism and oxidative profile: relevance for male fertility.

    PubMed

    Dias, Tânia R; Alves, Marco G; Bernardino, Raquel L; Martins, Ana D; Moreira, Ana C; Silva, Joaquina; Barros, Alberto; Sousa, Mário; Silva, Branca M; Oliveira, Pedro F

    2015-02-03

    Caffeine is a widely consumed substance present in several beverages. There is an increasing consumption of energetic drinks, rich in caffeine, among young individuals in reproductive age. Caffeine has been described as a modulator of cellular metabolism. Hence, we hypothesized that it alters human Sertoli cells (hSCs) metabolism and oxidative profile, which are essential for spermatogenesis. For that purpose, hSCs were cultured with increasing doses of caffeine (5, 50, 500 μM). Caffeine at the lowest concentrations (5 and 50 μM) stimulated lactate production, but only hSCs exposed to 50 μM showed increased expression of glucose transporters (GLUTs). At the highest concentration (500 μM), caffeine stimulated LDH activity to sustain lactate production. Notably, the antioxidant capacity of hSCs decreased in a dose-dependent manner and SCs exposed to 500 μM caffeine presented a pro-oxidant potential, with a concurrent increase of protein oxidative damage. Hence, moderate consumption of caffeine appears to be safe to male reproductive health since it stimulates lactate production by SCs, which can promote germ cells survival. Nevertheless, caution should be taken by heavy consumers of energetic beverages and food supplemented with caffeine to avoid deleterious effects in hSCs functioning and thus, abnormal spermatogenesis.

  18. Solar activity dependence of the topside ionosphere at low latitudes

    NASA Astrophysics Data System (ADS)

    Chen, Yiding; Liu, Libo; Wan, Weixing; Yue, Xinan; Su, Shin-Yi

    2009-08-01

    We investigated the solar activity dependence of the topside ionosphere with ROCSAT-1 observations. The distribution of the plasma density at 600 km altitude shows features with considerable local time, season, and solar activity differences. In the daytime, plasma density peaks around the dip equator. This peak is more distinct in equinoxes and weaker in May-July, and it enhances with solar activity in all seasons. The seasonal behavior of this peak is primarily controlled by the seasonal variations of neutral density and E × B vertical drift. The enhancement of the peak with solar activity is related to the effect of E × B vertical drift. Around sunset, double peaks are found in the latitudinal distribution of plasma density in solar maximum equinoxes and December solstice, which are mainly attributed to the effects of strong prereversal enhancement (PRE) vertical drift. Moreover, the plasma density at 600 km altitude strongly depends on the solar proxy P = (F 107 + F 107A)/2. At higher altitudes, e.g., 800 km, the amplification trend prevails in the solar activity variations of plasma density. In contrast, the plasma density at 600 km altitude presents three kinds of patterns (linear, amplification, and saturation), which has not been reported. Saturation effect is found at equinox sunset around the dip equator. This saturation effect is attributed to the increase in the PRE vertical drift with solar activity. Solar activity effects of ROCSAT-1 plasma density are argued to be the combined effects induced by the changes in the peak height, the scale height, and the peak electron density, respectively. Among these factors, the rise of the F 2 peak is more important for the equatorial plasma density at 600 km altitude.

  19. Bilateral Activity-Dependent Interactions in the Developing Corticospinal System

    PubMed Central

    Friel, Kathleen M.; Martin, John H.

    2009-01-01

    Activity-dependent competition between the corticospinal (CS) systems in each hemisphere drives postnatal development of motor skills and stable CS tract connections with contralateral spinal motor circuits. Unilateral restriction of motor cortex (M1) activity during an early postnatal critical period impairs contralateral visually guided movements later in development and in maturity. Silenced M1 develops aberrant connections with the contralateral spinal cord whereas the initially active M1, in the other hemisphere, develops bilateral connections. In this study, we determined whether the aberrant pattern of CS tract terminations and motor impairments produced by early postnatal M1 activity restriction could be abrogated by reducing activity-dependent synaptic competition from the initially active M1 later in development. We first inactivated M1 unilaterally between postnatal weeks 5–7. We next inactivated M1 on the other side from weeks 7–11 (alternate inactivation), to reduce the competitive advantage that this side may have over the initially inactivated side. Alternate inactivation redirected aberrant contralateral CS tract terminations from the initially silenced M1 to their normal spinal territories and reduced the density of aberrant ipsilateral terminations from the initially active side. Normal movement endpoint control during visually guided locomotion was fully restored. This reorganization of CS terminals reveals an unsuspected late plasticity after the critical period for establishing the pattern of CS terminations in the spinal cord. Our findings show that robust bilateral interactions between the developing CS systems on each side are important for achieving balance between contralateral and ipsilateral CS tract connections and visuomotor control. PMID:17928450

  20. Aven-dependent activation of ATM following DNA damage

    PubMed Central

    Guo, Jessie Yanxiang; Yamada, Ayumi; Kajino, Taisuke; Wu, Judy Qiju; Tang, Wanli; Freel, Christopher D.; Feng, Junjie; Chau, B. Nelson; Wang, Michael Zhuo; Margolis, Seth; Yoo, Hae Yong; Wang, Xiao-Fan; Dunphy, William G.; Irusta, Pablo M.; Hardwick, J. Marie; Kornbluth, Sally

    2009-01-01

    Summary Background In response to DNA damage, cells either undergo cell cycle arrest or apoptosis, depending on the extent of damage and the cell’s capacity for DNA repair. Cell cycle arrest induced by double-stranded DNA breaks depends on activation of the ataxia-telangiectasia (ATM) protein kinase, which phosphorylates cell cycle effectors such as Chk2 and p53 to inhibit cell cycle progression. ATM is recruited to double stranded DNA breaks by a complex of sensor proteins including Mre11/Rad50/Nbs1, resulting in autophosphorylation, monomerization, and activation of ATM kinase. Results In characterizing Aven protein, a previously reported apoptotic inhibitor, we have found that Aven can function as an ATM activator to inhibit G2/M progression. Aven bound to ATM and Aven overexpression in cycling Xenopus egg extracts prevented mitotic entry and induced phosphorylation of ATM and its substrates. Immunodepletion of endogenous Aven allowed mitotic entry even in the presence of damaged DNA, and RNAi-mediated knock-down of Aven in human cells prevented autophosphorylation of ATM at an activating site (S1981) in response to DNA damage. Interestingly, Aven is also a substrate of the ATM kinase. Mutation of ATM-mediated phosphorylation sites on Aven reduced its ability to activate ATM, suggesting that Aven activation of ATM following DNA damage is enhanced by ATM-mediated Aven phosphorylation. Conclusions These results identify Aven as a new ATM activator and describe a positive feedback loop operating between Aven and ATM. In aggregate, these findings place Aven, a known apoptotic inhibitor, as a critical transducer of the DNA damage signal. PMID:18571408

  1. A DNA enzyme with Mg(2+)-Dependent RNA Phosphoesterase Activity

    NASA Technical Reports Server (NTRS)

    Breaker, Ronald R.; Joyce, Gerald F.

    1995-01-01

    Previously we demonstrated that DNA can act as an enzyme in the Pb(2+)-dependent cleavage of an RNA phosphoester. This is a facile reaction, with an uncatalyzed rate for a typical RNA phosphoester of approx. 10(exp -4)/ min in the presence of 1 mM Pb(OAc)2 at pH 7.0 and 23 C. The Mg(2+) - dependent reaction is more difficult, with an uncatalyzed rate of approx. 10(exp -7)/ min under comparable conditions. Mg(2+) - dependent cleavage has special relevance to biology because it is compatible with intracellular conditions. Using in vitro selection, we sought to develop a family of phosphoester-cleaving DNA enzymes that operate in the presence of various divalent metals, focusing particularly on the Mg(2+) - dependent reaction. Results: We generated a population of greater than 10(exp 13) DNAs containing 40 random nucleotides and carried out repeated rounds of selective amplification, enriching for molecules that cleave a target RNA phosphoester in the presence of 1 mM Mg(2+), Mn(2+), Zn(2+) or Pb(2+). Examination of individual clones from the Mg(2+) lineage after the sixth round revealed a catalytic motif comprised of a three-stem junction.This motif was partially randomized and subjected to seven additional rounds of selective amplification, yielding catalysts with a rate of 0.01/ min. The optimized DNA catalyst was divided into separate substrate and enzyme domains and shown to have a similar level of activity under multiple turnover conditions. Conclusions: We have generated a Mg(2+) - dependent DNA enzyme that cleaves a target RNA phosphoester with a catalytic rate approx. 10(exp 5) - fold greater than that of the uncatalyzed reaction. This activity is compatible with intracellular conditions, raising the possibility that DNA enzymes might be made to operate in vivo.

  2. Direct evaluation of the position dependent diffusion coefficient and persistence time from the equilibrium density profile in anisotropic fluids.

    PubMed

    Olivares-Rivas, Wilmer; Colmenares, Pedro J; López, Floralba

    2013-08-21

    We derive expressions for the transverse diffusion coefficient D(z) and the average persistence time τ(z; L) within a layer of width L, for particles of a non-homogeneous fluid enclosed in a planar nanopore. The method allows the direct evaluation of these position-dependent dynamical quantities from the equilibrium local particle density profile. We use results for the density and persistence time profiles from the virtual layer molecular dynamics method to numerically assess the significance of the Smoluchowski approximation.

  3. Activity-dependent, homeostatic regulation of neurotransmitter release from auditory nerve fibers

    PubMed Central

    Ngodup, Tenzin; Goetz, Jack A.; McGuire, Brian C.; Sun, Wei; Lauer, Amanda M.; Xu-Friedman, Matthew A.

    2015-01-01

    Information processing in the brain requires reliable synaptic transmission. High reliability at specialized auditory nerve synapses in the cochlear nucleus results from many release sites (N), high probability of neurotransmitter release (Pr), and large quantal size (Q). However, high Pr also causes auditory nerve synapses to depress strongly when activated at normal rates for a prolonged period, which reduces fidelity. We studied how synapses are influenced by prolonged activity by exposing mice to constant, nondamaging noise and found that auditory nerve synapses changed to facilitating, reflecting low Pr. For mice returned to quiet, synapses recovered to normal depression, suggesting that these changes are a homeostatic response to activity. Two additional properties, Q and average excitatory postsynaptic current (EPSC) amplitude, were unaffected by noise rearing, suggesting that the number of release sites (N) must increase to compensate for decreased Pr. These changes in N and Pr were confirmed physiologically using the integration method. Furthermore, consistent with increased N, endbulbs in noise-reared animals had larger VGlut1-positive puncta, larger profiles in electron micrographs, and more release sites per profile. In current-clamp recordings, noise-reared BCs had greater spike fidelity even during high rates of synaptic activity. Thus, auditory nerve synapses regulate excitability through an activity-dependent, homeostatic mechanism, which could have major effects on all downstream processing. Our results also suggest that noise-exposed bushy cells would remain hyperexcitable for a period after returning to normal quiet conditions, which could have perceptual consequences. PMID:25944933

  4. Role of BDNF epigenetics in activity-dependent neuronal plasticity.

    PubMed

    Karpova, Nina N

    2014-01-01

    Brain-derived neurotrophic factor (BDNF) is a key mediator of the activity-dependent processes in the brain that have a major impact on neuronal development and plasticity. Impaired control of neuronal activity-induced BDNF expression mediates the pathogenesis of various neurological and psychiatric disorders. Different environmental stimuli, such as the use of pharmacological compounds, physical and learning exercises or stress exposure, lead to activation of specific neuronal networks. These processes entail tight temporal and spatial transcriptional control of numerous BDNF splice variants through epigenetic mechanisms. The present review highlights recent findings on the dynamic and long-term epigenetic programming of BDNF gene expression by the DNA methylation, histone-modifying and microRNA machineries. The review also summarizes the current knowledge on the activity-dependent BDNF mRNA trafficking critical for rapid local regulation of BDNF levels and synaptic plasticity. Current data open novel directions for discovery of new promising therapeutic targets for treatment of neuropsychiatric disorders. This article is part of the Special Issue entitled 'BDNF Regulation of Synaptic Structure, Function, and Plasticity'. Copyright © 2013 Elsevier Ltd. All rights reserved.

  5. Catalytic reaction profile for NADH-dependent reduction of aromatic aldehydes by xylose reductase from Candida tenuis.

    PubMed Central

    Mayr, Peter; Nidetzky, Bernd

    2002-01-01

    Kinetic substituent effects have been used to examine the catalytic reaction profile of xylose reductase from the yeast Candida tenuis, a representative aldo/keto reductase of primary carbohydrate metabolism. Michaelis-Menten parameters (k(cat) and K(m)) for NADH-dependent enzymic aldehyde reductions have been determined using a homologous series of benzaldehyde derivatives in which substituents in meta and para positions were employed to systematically perturb the properties of the reactive carbonyl group. Kinetic isotope effects (KIEs) on k(cat) and k(cat)/K(m) for enzymic reactions with meta-substituted benzaldehydes have been obtained by using NADH (2)H-labelled in the pro-R C4-H position, and equilibrium constants for the conversion of these aldehydes into the corresponding alcohols (K(eq)) have been measured in the presence of NAD(H) and enzyme. Aldehyde dissociation constants (K(d)) and the hydride transfer rate constant (k(7)) have been calculated from steady-state rate and KIE data. Quantitative structure-activity relationship analysis was used to factor the observed substituent dependence of k(cat)/K(m) into a major electronic effect and a productive positional effect of the para substituent. k(cat)/K(m) (after correction for substituent position) and K(eq) obeyed log-linear correlations over the substituent parameter, Hammett sigma, giving identical slope values (rho) of +1.4 to +1.7, whereas the same Hammett plot for logK(d) yielded rho=-1.5. This leads to the conclusion that electron-withdrawing substituents facilitate the reaction and increase binding to about the same extent. KIE values for k(cat) (1.8) and k(cat)/K(m) (2.7), and likewise k(7), showed no substituent dependence. Therefore, irrespective of the observed changes in reactivity over the substrate series studied no shift in the character of the rate-limiting transition state of hydride transfer occurred. The signs and magnitudes of rho values suggest this transition state to be product

  6. The quality of feedback during formative OSCEs depends on the tutors' profile.

    PubMed

    Junod Perron, Noelle; Louis-Simonet, Martine; Cerutti, Bernard; Pfarrwaller, Eva; Sommer, Johanna; Nendaz, Mathieu

    2016-11-15

    During their pre-clinical years, medical students are given the opportunity to practice clinical skills with simulated patients. During these formative objective structured clinical encounters (OSCEs), tutors from various backgrounds give feedback on students' history taking, physical exam, and communication skills. The aim of the study was to evaluate whether the content and process of feedback varied according to the tutors' profile. During 2013, all 2(nd) and 3(rd) year medical students and tutors involved in three formative OSCEs were asked to fill in questionnaires, and their feedback sessions were audiotaped. Tutors were divided into two groups: 1) generalists: primary care, general internist and educationalist physicians 2) specialists involved in the OSCE related to their field of expertise. Outcome measures included the students' perceptions of feedback quality and utility and objective assessment of feedback quality. Participants included 251 medical students and 38 tutors (22 generalists and 16 specialists). Students self-reported that feedback was useful to improve history taking, physical exam and communication skills. Objective assessment showed that feedback content essentially focused on history taking and physical exam skills, and that elaboration on clinical reasoning or communication/professionalism issues was uncommon. Multivariate analyses showed that generalist tutors used more learner-centered feedback skills than specialist tutors (stimulating student's self-assessment (p < .001; making the student active in finding solutions, p < .001; checking student's understanding, p < .001) and elaborated more on communication and professionalism issues (p < 0.001). Specialists reported less training in how to provide feedback than generalists. These findings suggest that generalist tutors are more learner-centered and pay more attention to communication and professionalism during feedback than specialist tutors. Such differences may be

  7. Deciphering Phosphotyrosine-Dependent Signaling Networks in Cancer by SH2 Profiling

    PubMed Central

    Machida, Kazuya; Khenkhar, Malik

    2012-01-01

    It has been a decade since the introduction of SH2 profiling, a modular domain-based molecular diagnostics tool. This review covers the original concept of SH2 profiling, different analytical platforms, and their applications, from the detailed analysis of single proteins to broad screening in translational research. Illustrated by practical examples, we discuss the uniqueness and advantages of the approach as well as its limitations and challenges. We provide guidance for basic researchers and oncologists who may consider SH2 profiling in their respective cancer research, especially for those focusing on tyrosine phosphoproteomics. SH2 profiling can serve as an alternative phosphoproteomics tool to dissect aberrant tyrosine kinase pathways responsible for individual malignancies, with the goal of facilitating personalized diagnostics for the treatment of cancer. PMID:23226573

  8. Constructing bioactive peptides with pH-dependent activities.

    PubMed

    Tu, Zhigang; Volk, Melanie; Shah, Khushali; Clerkin, Kevin; Liang, Jun F

    2009-08-01

    Many bioactive peptides are featured by their arginine and lysine rich contents. In this study, lysine and arginine residues in lytic peptides were selectively replaced by histidines. Although resulting histidine-containing lytic peptides had decreased activity, they did show pH-dependent cytotoxicity. The activity of the constructed histidine-containing lytic peptides increased 2-8 times as the solution pH changed from 7.4 to 5.5. More importantly, these histidine-containing peptides maintain the same cell killing mechanism as their parent peptides by causing cell lysis. Both the activity and pH-sensitivity of histidine-containing peptides are tunable by adjusting histidine substitution numbers and positions. This study has presented a general strategy to create bioactive peptides with desired pH-sensitivity to meet the needs of various applications such as cancer treatments.

  9. Constructing bioactive peptides with pH-dependent activities

    PubMed Central

    Tu, Zhigang; Volk, Melanie; Shah, Khushali; Clerkin, Kevin; Liang, Jun F.

    2009-01-01

    Many bioactive peptides are featured by their arginine and lysine rich contents. In this study, lysine and arginine residues in lytic peptides were selectively replaced by histidines. Although resulted histidine-containing lytic peptides had decreased activity, they did show pH-dependent cytotoxicity. The activity of the constructed histidine-containing lytic peptides increased 2 ~ 8 times as the solution pH changed from of 7.4 to 5.5. More importantly, these histidine-containing peptides maintain the same cell killing mechanism as their parent peptides by causing cell lysis. Both the activity and pH-sensitivity of histidine-containing peptides are tunable by adjusting histidine substitution numbers and positions. This study has presented a general strategy to create bioactive peptides with desired pH-sensitivity to meet the needs of various applications such as cancer treatments. PMID:19464332

  10. Neuronal activity-dependent membrane traffic at the neuromuscular junction

    PubMed Central

    Miana-Mena, Francisco Javier; Roux, Sylvie; Benichou, Jean-Claude; Osta, Rosario; Brûlet, Philippe

    2002-01-01

    During development and also in adulthood, synaptic connections are modulated by neuronal activity. To follow such modifications in vivo, new genetic tools are designed. The nontoxic C-terminal fragment of tetanus toxin (TTC) fused to a reporter gene such as LacZ retains the retrograde and transsynaptic transport abilities of the holotoxin itself. In this work, the hybrid protein is injected intramuscularly to analyze in vivo the mechanisms of intracellular and transneuronal traffics at the neuromuscular junction (NMJ). Traffic on both sides of the synapse are strongly dependent on presynaptic neural cell activity. In muscle, a directional membrane traffic concentrates β-galactosidase-TTC hybrid protein into the NMJ postsynaptic side. In neurons, the probe is sorted across the cell to dendrites and subsequently to an interconnected neuron. Such fusion protein, sensitive to presynaptic neuronal activity, would be extremely useful to analyze morphological changes and plasticity at the NMJ. PMID:11880654

  11. Plasminogen-dependent proteolytic activity in Bifidobacterium lactis.

    PubMed

    Candela, Marco; Miccoli, Giacomo; Bergmann, Simone; Turroni, Silvia; Vitali, Beatrice; Hammerschmidt, Sven; Brigidi, Patrizia

    2008-08-01

    Bifidobacteria represent one of the most important health-promoting bacterial groups of the intestinal microbiota. The binding of plasminogen to species of Bifidobacterium has been recently reported. To further explore the interaction between bifidobacteria and plasminogen, we investigated the role of Bifidobacterium lactis BI07 plasminogen-dependent proteolytic activity in the degradation of host-specific substrates. Our experimental data demonstrate that the recruitment of plasminogen on the bacterial cell surface and its subsequent conversion into plasmin by host-derived plasminogen activators provide B. lactis BI07 with a surface-associated plasmin activity effective in degradation of physiological substrates such as extracellular matrix, fibronectin and fibrinogen. The ability of bifidobacteria to intervene in the host plasminogen/plasmin system may contribute to facilitating colonization of the host gastrointestinal tract.

  12. Altered Spontaneous Brain Activity in Betel Quid Dependence

    PubMed Central

    Liu, Tao; Li, Jian-jun; Zhao, Zhong-yan; Yang, Guo-shuai; Pan, Meng-jie; Li, Chang-qing; Pan, Su-yue; Chen, Feng

    2016-01-01

    Abstract It has been suggested by the first voxel-based morphometry investigation that betel quid dependence (BQD) individuals are presented with brain structural changes in previous reports, and there may be a neurobiological basis for BQD individuals related to an increased risk of executive dysfunction and disinhibition, subjected to the reward system, cognitive system, and emotion system. However, the effects of BQD on neural activity remain largely unknown. Individuals with impaired cognitive control of behavior often reveal altered spontaneous cerebral activity in resting-state functional magnetic resonance imaging and those changes are usually earlier than structural alteration. Here, we examined BQD individuals (n = 33) and age-, sex-, and education-matched healthy control participants (n = 32) in an resting-state functional magnetic resonance imaging study to observe brain function alterations associated with the severity of BQD. Amplitude of low-frequency fluctuation (ALFF) and regional homogeneity (ReHo) values were both evaluated to stand for spontaneous cerebral activity. Gray matter volumes of these participants were also calculated for covariate. In comparison with healthy controls, BQD individuals demonstrated dramatically decreased ALFF and ReHo values in the prefrontal gurus along with left fusiform, and increased ALFF and ReHo values in the primary motor cortex area, temporal lobe as well as some regions of occipital lobe. The betel quid dependence scores (BQDS) were negatively related to decreased activity in the right anterior cingulate. The abnormal spontaneous cerebral activity revealed by ALFF and ReHo calculation excluding the structural differences in patients with BQD may help us probe into the neurological pathophysiology underlying BQD-related executive dysfunction and disinhibition. Diminished spontaneous brain activity in the right anterior cingulate cortex may, therefore, represent a biomarker of BQD individuals. PMID

  13. Functionally diverse biotin-dependent enzymes with oxaloacetate decarboxylase activity.

    PubMed

    Lietzan, Adam D; St Maurice, Martin

    2014-02-15

    Biotin-dependent enzymes catalyze carboxylation, decarboxylation and transcarboxylation reactions that participate in the primary metabolism of a wide range of organisms. In all cases, the overall reaction proceeds via two half reactions that take place in physically distinct active sites. In the first half-reaction, a carboxyl group is transferred to the 1-N' of a covalently tethered biotin cofactor. The tethered carboxybiotin intermediate subsequently translocates to a second active site where the carboxyl group is either transferred to an acceptor substrate or, in some bacteria and archaea, is decarboxylated to biotin and CO2 in order to power the export of sodium ions from the cytoplasm. A homologous carboxyltransferase domain is found in three enzymes that catalyze diverse overall reactions: carbon fixation by pyruvate carboxylase, decarboxylation and sodium transport by the biotin-dependent oxaloacetate decarboxylase complex, and transcarboxylation by transcarboxylase from Propionibacterium shermanii. Over the past several years, structural data have emerged which have greatly advanced the mechanistic description of these enzymes. This review assembles a uniform description of the carboxyltransferase domain structure and catalytic mechanism from recent studies of pyruvate carboxylase, oxaloacetate decarboxylase and transcarboxylase, three enzymes that utilize an analogous carboxyltransferase domain to catalyze the biotin-dependent decarboxylation of oxaloacetate.

  14. E region electric field dependence of the solar activity

    NASA Astrophysics Data System (ADS)

    Denardini, C. M.; Moro, J.; Resende, L. C. A.; Chen, S. S.; Schuch, N. J.; Costa, J. E. R.

    2015-10-01

    We have being studying the zonal and vertical E region electric field components inferred from the Doppler shifts of type 2 echoes (gradient drift irregularities) detected with the 50 MHz backscatter coherent radar set at São Luis, Brazil (SLZ, 2.3°S, 44.2°W) during the solar cycle 24. In this report we present the dependence of the vertical and zonal components of this electric field with the solar activity, based on the solar flux F10.7. For this study we consider the geomagnetically quiet days only (Kp ≤ 3+). A magnetic field-aligned-integrated conductivity model was developed for proving the conductivities, using the IRI-2007, the MISIS-2000, and the IGRF-11 models as input parameters for ionosphere, neutral atmosphere, and Earth magnetic field, respectively. The ion-neutron collision frequencies of all the species are combined through the momentum transfer collision frequency equation. The mean zonal component of the electric field, which normally ranged from 0.19 to 0.35 mV/m between the 8 and 18 h (LT) in the Brazilian sector, show a small dependency with the solar activity. Whereas the mean vertical component of the electric field, which normally ranges from 4.65 to 10.12 mV/m, highlights the more pronounced dependency of the solar flux.

  15. A total ozone-dependent ozone profile climatology based on ozonesondes and Aura MLS data

    NASA Astrophysics Data System (ADS)

    Labow, Gordon J.; Ziemke, Jerald R.; McPeters, Richard D.; Haffner, David P.; Bhartia, Pawan K.

    2015-03-01

    Ozone profiles measured with the Aura Microwave Limb Sounder (MLS) and ozonesondes are used to create a new ozone climatology that can be used for satellite retrievals and radiative transfer studies. The climatology is binned according to total column ozone amount and latitude rather than with season. Because of high correlation between ozone profile shape and total ozone, the ozone profiles in this climatology capture ozone variations well, especially near the tropopause. This climatology has been constructed from nearly a million individual MLS ozone profile measurements taken between 2004 and 2013 as well as over 55,000 ozonesonde measurements from 1988 to 2011. The MLS profiles were sorted by total column ozone as measured by Ozone Monitoring Instrument in observations that were coincident with the MLS measurements. The data from the sondes were used in the troposphere and lower stratosphere and MLS in the middle and upper stratosphere. These two data sets were blended together between 13 and 17 km (~159-88 hPa). This climatology consists of average ozone profiles as a function of total ozone for six 30° latitude bands covering altitudes between 0 and 75 km (in Z* pressure altitude coordinates) as well as the corresponding standard deviations for each layer. There is no seasonal component. This new climatology shows some remarkable and somewhat unexpected correlations between the total column ozone and the ozone amount at some layers, particularly in the lower and middle troposphere in some latitude bands.

  16. Protein profiling of mouse livers with peroxisome proliferator-activated receptor alpha activation.

    PubMed

    Chu, Ruiyin; Lim, Hanjo; Brumfield, Laura; Liu, Hong; Herring, Chris; Ulintz, Peter; Reddy, Janardan K; Davison, Matthew

    2004-07-01

    Peroxisome proliferator-activated receptor alpha (PPARalpha) is important in the induction of cell-specific pleiotropic responses, including the development of liver tumors, when it is chronically activated by structurally diverse synthetic ligands such as Wy-14,643 or by unmetabolized endogenous ligands resulting from the disruption of the gene encoding acyl coenzyme A (CoA) oxidase (AOX). Alterations in gene expression patterns in livers with PPARalpha activation were delineated by using a proteomic approach to analyze liver proteins of Wy-14,643-treated and AOX(-/-) mice. We identified 46 differentially expressed proteins in mouse livers with PPARalpha activation. Up-regulated proteins, including acetyl-CoA acetyltransferase, farnesyl pyrophosphate synthase, and carnitine O-octanoyltransferase, are involved in fatty acid metabolism, whereas down-regulated proteins, including ketohexokinase, formiminotransferase-cyclodeaminase, fructose-bisphosphatase aldolase B, sarcosine dehydrogenase, and cysteine sulfinic acid decarboxylase, are involved in carbohydrate and amino acid metabolism. Among stress response and xenobiotic metabolism proteins, selenium-binding protein 2 and catalase showed a dramatic approximately 18-fold decrease in expression and a modest approximately 6-fold increase in expression, respectively. In addition, glycine N-methyltransferase, pyrophosphate phosphohydrolase, and protein phosphatase 1D were down-regulated with PPARalpha activation. These observations establish proteomic profiles reflecting a common and predictable pattern of differential protein expression in livers with PPARalpha activation. We conclude that livers with PPARalpha activation are transcriptionally geared towards fatty acid combustion.

  17. Activity-dependent plasticity of mouse hippocampal assemblies in vitro

    PubMed Central

    Keller, Martin K.; Draguhn, Andreas; Both, Martin; Reichinnek, Susanne

    2015-01-01

    Memory formation is associated with the generation of transiently stable neuronal assemblies. In hippocampal networks, such groups of functionally coupled neurons express highly ordered spatiotemporal activity patterns which are coordinated by local network oscillations. One of these patterns, sharp wave-ripple complexes (SPW-R), repetitively activates previously established groups of memory-encoding neurons, thereby supporting memory consolidation. This function implies that repetition of specific SPW-R induces plastic changes which render the underlying neuronal assemblies more stable. We modeled this repetitive activation in an in vitro model of SPW-R in mouse hippocampal slices. Weak electrical stimulation upstream of the CA3-CA1 networks reliably induced SPW-R of stereotypic waveform, thus representing re-activation of similar neuronal activity patterns. Frequent repetition of these patterns (100 times) reduced the variance of both, evoked and spontaneous SPW-R waveforms, indicating stabilization of pre-existing assemblies. These effects were most pronounced in the CA1 subfield and depended on the timing of stimulation relative to spontaneous SPW-R. Additionally, plasticity of SPW-R was blocked by application of a NMDA receptor antagonist, suggesting a role for associative synaptic plasticity in this process. Thus, repetitive activation of specific patterns of SPW-R causes stabilization of memory-related networks. PMID:26041998

  18. Brain activation during sight gags and language-dependent humor.

    PubMed

    Watson, Karli K; Matthews, Benjamin J; Allman, John M

    2007-02-01

    Humor is a hallmark of human discourse. People use it to relieve stress and to facilitate social bonding, as well as for pure enjoyment in the absence of any apparent adaptive value. Although recent studies have revealed that humor acts as an intrinsic reward, which explains why people actively seek to experience and create humor, few have addressed the cognitive aspects of humor. We used event-related functional magnetic resonance imaging to differentiate brain activity induced by the hedonic similarities and cognitive differences inherent in 2 kinds of humor: visual humor (sight gags) and language-based humor. Our findings indicate that the brain networks recruited during a humorous experience differ according to the type of humor being processed, with high-level visual areas activated during visual humor and classic language areas activated during language-dependent humor. Our results additionally highlight a common network activated by both types of humor that includes the amygdalar and midbrain regions, which presumably reflect the euphoric component of humor. Furthermore, we found that humor activates anterior cingulate cortex and frontoinsular cortex, 2 regions in the brain that are known to have phylogenetically recent neuronal circuitry. These results suggest that humor may have coevolved with another cognitive specialization of the great apes and humans: the ability to navigate through a shifting and complex social space.

  19. Quantifying the fingerprint descriptor dependence of structure-activity relationship information on a large scale.

    PubMed

    Dimova, Dilyana; Stumpfe, Dagmar; Bajorath, Jürgen

    2013-09-23

    It is well-known that different molecular representations, e.g., graphs, numerical descriptors, fingerprints, or 3D models, change the numerical results of molecular similarity calculations. Because the assessment of structure-activity relationships (SARs) requires similarity and potency comparisons of active compounds, this representation dependence inevitably also affects SAR analysis. But to what extent? How exactly does SAR information change when alternative fingerprints are used as descriptors? What is the proportion of active compounds with substantial changes in SAR information induced by different fingerprints? To provide answers to these questions, we have quantified changes in SAR information across many different compound classes using six different fingerprints. SAR profiling was carried out on 128 target-based data sets comprising more than 60,000 compounds with high-confidence activity annotations. A numerical measure of SAR discontinuity was applied to assess SAR information on a per compound basis. For ~70% of all test compounds, changes in SAR characteristics were detected when different fingerprints were used as molecular representations. Moreover, the SAR phenotype of ~30% of the compounds changed, and distinct fingerprint-dependent local SAR environments were detected. The fingerprints we compared were found to generate SAR models that were essentially not comparable. Atom environment and pharmacophore fingerprints produced the largest differences in compound-associated SAR information. Taken together, the results of our systematic analysis reveal larger fingerprint-dependent changes in compound-associated SAR information than would have been anticipated.

  20. A microfluidic device for evaluating the dynamics of the metabolism-dependent antioxidant activity of nutrients.

    PubMed

    Lee, Jungwoo; Choi, Jong-ryul; Ha, Sang Keun; Choi, Inwook; Lee, Seung Hwan; Kim, Donghyun; Choi, Nakwon; Sung, Jong Hwan

    2014-08-21

    Various food components are known for their health-promoting effects. However, their biochemical effects are generally evaluated in vitro, and their actual in vivo effect can vary significantly, depending on their metabolic profiles. To evaluate the effect of the liver metabolism on the antioxidant activity, we have developed a two-compartment microfluidic system that integrates the dynamics of liver metabolism and the subsequent antioxidant activity of food components. In the first compartment of the device, human liver enzyme fractions were immobilized inside a poly(ethylene glycol) diacrylate (PEGDA) hydrogel to mimic the liver metabolism. The radical scavenging activity was evaluated by the change of the 2,2-diphenyl-1-picrylhydrazyl (DPPH) absorbance in the second compartment. Reaction engineering and fluid mechanics principles were used to develop a simplified analytical model and a more complex finite element model, which were used to design the chip and determine the optimal flow conditions. For real-time measurements of the reaction on a chip, we developed a custom-made photospectrometer system with an LED light source. The developed microfluidic system showed a linear and dose-dependent antioxidant activity in response to increasing concentration of flavonoid. We also compared the antioxidant activity of flavonoid after various liver metabolic reactions. This microfluidic system can serve as a novel in vitro platform for predicting the antioxidant activity of various food components in a more physiologically realistic manner, as well as for studying the mechanism of action of such food components.

  1. Structural basis for proteolysis-dependent activation of the poliovirus RNA-dependent RNA polymerase

    PubMed Central

    Thompson, Aaron A; Peersen, Olve B

    2004-01-01

    The active RNA-dependent RNA polymerase of poliovirus, 3Dpol, is generated by cleavage of the 3CDpro precursor protein, a protease that has no polymerase activity despite containing the entire polymerase domain. By intentionally disrupting a known and persistent crystal packing interaction, we have crystallized the poliovirus polymerase in a new space group and solved the complete structure of the protein at 2.0 Å resolution. It shows that the N-terminus of fully processed 3Dpol is buried in a surface pocket where it makes hydrogen bonds that act to position Asp238 in the active site. Asp238 is an essential residue that selects for the 2′ OH group of substrate rNTPs, as shown by a 2.35 Å structure of a 3Dpol–GTP complex. Mutational, biochemical, and structural data further demonstrate that 3Dpol activity is exquisitely sensitive to mutations at the N-terminus. This sensitivity is the result of allosteric effects where the structure around the buried N-terminus directly affects the positioning of Asp238 in the active site. PMID:15306852

  2. pH dependence on functional activity of human and mouse flavin-containing monooxygenase 5.

    PubMed

    Motika, Meike S; Zhang, Jun; Ralph, Erik C; Dwyer, Mary A; Cashman, John R

    2012-04-01

    Flavin-containing monooxygenase (FMO) 5 belongs to a family of enzymes that catalyze the oxygenation of nucleophilic N- and S-containing compounds. The FMO enzyme family consists of five forms (FMOs1-5) that share about 50-60% sequence identity to each other. A comparison of FMOs showed that the pH-dependence profile for functional activity of FMO5 differed significantly from that of other FMO enzymes. The objective of this study was to examine the pH-dependence of FMO5 to gain insight into the mechanism of action of FMO5. Recombinant mouse and human FMO5 (mFMO5 and hFMO5, respectively) were expressed as maltose-binding fusion proteins from Escherichia coli, purified with affinity chromatography, and examined for their N-oxygenation functional activity at different pH values. hFMO5 showed a broader range and greater functional activity from pH 6 to 11 compared to mFMO5. mFMO5 lost almost all functional activity at pH 6, while hFMO5 maintained almost normal enzyme activity. In order to identify the amino acid residues involved in the effects of pH on hFMO5 and mFMO5 functional enzyme activity, pH-studies in the range of pH 6-9 were done with chimeras of recombinant mouse and human FMO5 and variants of both. Results of these studies and molecular modeling showed that residues responsible for the differences in the pH profile between mFMO5 and hFMO5 were located at positions 227 and 228 of the enzyme. Further variants were made to investigate the role of these amino acids. The results of this study may help to explain the mechanism of FMO function. Copyright © 2012 Elsevier Inc. All rights reserved.

  3. Derivation of the Fano profile from time-dependent density-functional theory for local thermodynamic equilibrium plasmas

    NASA Astrophysics Data System (ADS)

    Kiyokawa, Shuji

    2007-04-01

    We give the derivation of the Fano profile (the resonance energy position, the resonance width Γ , and q value) from the time-dependent nonrelativistic density-functional theory (DFT) and propose a scheme for calculating the photoabsorption cross section of hot dense plasmas. As a consequence of this derivation, we show the line profile is obtained as a superposition of Fano and Lorentz profiles when the competition of two optically allowed bound-bound and bound-free transitions occurs. We also show the results of the photoabsorption cross section by applying our scheme to an Fe plasma (density is 7.85g/cm3 , temperature is 100eV ), where the calculation is carried out without numerical divergence for any photon energy. The calculated results are in good agreement with those of Grimaldi.

  4. Leukocyte and plasma activation profiles in chronically transfused patients with a history of allergic reactions.

    PubMed

    Fontaine, Magali J; Shih, Hank; Schubert, Richard; Wong, Wendy; Andrews, Jennifer; Jeng, Michael; Tirouvanziam, Rabindra

    2017-09-06

    Allergic transfusion reactions are drawbacks to the benefits of transfusion. Classically, allergic transfusion reactions depend on histamine release from mast cells or basophils, but other leukocyte subsets may also be important. Thus, we propose to better define the exact leukocyte subsets involved in allergic transfusion reactions. The overall objective of the current study was to compare the activation of specific peripheral blood leukocyte subsets (monocytes, neutrophils, eosinophils, and basophils) in a cohort of 13 patients who received chronic transfusions and had a history of allergic transfusion reactions compared with a control group of patients who received chronic transfusions and had no history of allergic transfusion reactions. Leukocyte subsets were analyzed by flow cytometry at baseline and after red blood cell transfusion, and cytokine levels in platelet-free plasma collected at the same time points were measured by Luminex assay. Flow cytometry and cytokine profiles before and after transfusion did not differ significantly between patients who did and did not have a history of allergic transfusion reactions (p > 0.05). However, post-transfusion samples from both groups showed a decrease in CD63 expression in basophils, monocytes, and eosinophils and a decrease in CD45 expression in all leukocyte subsets compared with pretransfusion samples. Interleukin 10 levels increased after transfusion in the group with a history of allergic transfusion reactions (p = 0.0469), and RANTES (regulated upon activation, normal T-cell expressed and secreted) was significantly decreased post-transfusion in all patients (p = 0.0122). None of the leukocyte subsets from patients who had a history of allergic transfusion reactions significantly increased in activation either before or after transfusion. All leukocyte subsets from patients who did and did not have a history of allergic transfusion reactions decreased in their activation profile upon

  5. Regional variation in myofilament length-dependent activation.

    PubMed

    Cazorla, Olivier; Lacampagne, Alain

    2011-07-01

    The Frank-Starling law is an important regulatory mechanism of the heart that links the end-diastolic volume with the systolic ejection fraction. This beat-to-beat regulation of the heart, underlined at the cellular level by higher myofilament calcium sensitivity at longer sarcomere length, is known as length-dependent activation or stretch sensitization of activation. However, the heart is structurally and functionally heterogeneous and asymmetrical. Specifically, contractile properties are not uniform within the left ventricle partly due to transmural differences in action potential waveforms and calcium homeostasis. The present review will focus on the role of the contractile machinery in the transmural contractile heterogeneity and its adaptation to changes in muscle strain. The expression of different myosin isoforms, the level of titin-based passive tension, and thin and thick sarcomeric regulatory proteins are considered to explain the regional cellular contractile properties. Finally, the importance of transmural heterogeneity of length-dependent activation and the consequences of its modification on the heart mechanics are discussed. Despite extensive research since the characterization of the Frank-Starling law, the molecular mechanisms by which strain information is transduced to the contractile machinery have not been fully determined yet.

  6. Nitric oxide mediates local activity-dependent excitatory synapse development.

    PubMed

    Nikonenko, Irina; Nikonenko, Alexander; Mendez, Pablo; Michurina, Tatyana V; Enikolopov, Grigori; Muller, Dominique

    2013-10-29

    Learning related paradigms play an important role in shaping the development and specificity of synaptic networks, notably by regulating mechanisms of spine growth and pruning. The molecular events underlying these synaptic rearrangements remain poorly understood. Here we identify NO signaling as a key mediator of activity-dependent excitatory synapse development. We find that chronic blockade of NO production in vitro and in vivo interferes with the development of hippocampal and cortical excitatory spine synapses. The effect results from a selective loss of activity-mediated spine growth mechanisms and is associated with morphological and functional alterations of remaining synapses. These effects of NO are mediated by a cGMP cascade and can be reproduced or prevented by postsynaptic expression of vasodilator-stimulated phosphoprotein phospho-mimetic or phospho-resistant mutants. In vivo analyses show that absence of NO prevents the increase in excitatory synapse density induced by environmental enrichment and interferes with the formation of local clusters of excitatory synapses. We conclude that NO plays an important role in regulating the development of excitatory synapses by promoting local activity-dependent spine-growth mechanisms.

  7. Retinal Wave Behavior through Activity-Dependent Refractory Periods

    PubMed Central

    Godfrey, Keith B; Swindale, Nicholas V

    2007-01-01

    In the developing mammalian visual system, spontaneous retinal ganglion cell (RGC) activity contributes to and drives several aspects of visual system organization. This spontaneous activity takes the form of spreading patches of synchronized bursting that slowly advance across portions of the retina. These patches are non-repeating and tile the retina in minutes. Several transmitter systems are known to be involved, but the basic mechanism underlying wave production is still not well-understood. We present a model for retinal waves that focuses on acetylcholine mediated waves but whose principles are adaptable to other developmental stages. Its assumptions are that a) spontaneous depolarizations of amacrine cells drive wave activity; b) amacrine cells are locally connected, and c) cells receiving more input during their depolarization are subsequently less responsive and have longer periods between spontaneous depolarizations. The resulting model produces waves with non-repeating borders and randomly distributed initiation points. The wave generation mechanism appears to be chaotic and does not require neural noise to produce this wave behavior. Variations in parameter settings allow the model to produce waves that are similar in size, frequency, and velocity to those observed in several species. Our results suggest that retinal wave behavior results from activity-dependent refractory periods and that the average velocity of retinal waves depends on the duration a cell is excitatory: longer periods of excitation result in slower waves. In contrast to previous studies, we find that a single layer of cells is sufficient for wave generation. The principles described here are very general and may be adaptable to the description of spontaneous wave activity in other areas of the nervous system. PMID:18052546

  8. Activation of cyclin A-dependent protein kinases during apoptosis.

    PubMed Central

    Meikrantz, W; Gisselbrecht, S; Tam, S W; Schlegel, R

    1994-01-01

    Apoptosis was induced in S-phase-arrested HeLa cells by staurosporine, caffeine, 6-dimethylaminopurine, and okadaic acid, agents that activate M-phase-promoting factor and induce premature mitosis in similarly treated hamster cell lines. Addition of these agents to asynchronously growing HeLa cells or to cells arrested in early G1 phase with lovastatin had little or no effect. S-phase arrest also promoted tumor necrosis factor alpha-induced apoptosis, eliminating the normal requirement for simultaneous cycloheximide treatment. For all of the apoptosis-inducing agents tested, the appearance of condensed chromatin was accompanied by 2- to 7-fold increases in cyclin A-associated histone H1 kinase activity, levels approximating the mitotic value. Where examined, both Cdc2 and Cdk2, the catalytic subunits known to associate with cyclin A, were activated. Stable overexpression of bcl-2 suppressed the apoptosis-inducing activity of all agents tested and reduced the amount of Cdc2 and Cdk2 in the nucleus, suggesting a possible mechanism by which bcl-2 inhibits the chromatin condensation characteristic of apoptosis. These findings suggest that at least one of the biochemical steps required for mitosis, activation of cyclin A-dependent protein kinases, is also an important event during apoptosis. Images PMID:8170983

  9. Shaping inhibition: activity dependent structural plasticity of GABAergic synapses

    PubMed Central

    Flores, Carmen E.; Méndez, Pablo

    2014-01-01

    Inhibitory transmission through the neurotransmitter γ-aminobutyric acid (GABA) shapes network activity in the mammalian cerebral cortex by filtering synaptic incoming information and dictating the activity of principal cells. The incredibly diverse population of cortical neurons that use GABA as neurotransmitter shows an equally diverse range of mechanisms that regulate changes in the strength of GABAergic synaptic transmission and allow them to dynamically follow and command the activity of neuronal ensembles. Similarly to glutamatergic synaptic transmission, activity-dependent functional changes in inhibitory neurotransmission are accompanied by alterations in GABAergic synapse structure that range from morphological reorganization of postsynaptic density to de novo formation and elimination of inhibitory contacts. Here we review several aspects of structural plasticity of inhibitory synapses, including its induction by different forms of neuronal activity, behavioral and sensory experience and the molecular mechanisms and signaling pathways involved. We discuss the functional consequences of GABAergic synapse structural plasticity for information processing and memory formation in view of the heterogenous nature of the structural plasticity phenomena affecting inhibitory synapses impinging on somatic and dendritic compartments of cortical and hippocampal neurons. PMID:25386117

  10. relA-dependent RNA polymerase activity in Escherichia coli.

    PubMed Central

    Ryals, J; Bremer, H

    1982-01-01

    Parameters relating to RNA synthesis were measured after a temperature shift from 30 to 42 degrees C, in a relA+ and relA- isogenic pair of Escherichia coli strains containing a temperature-sensitive valyl tRNA synthetase. The following results were obtained: (i) the rRNA chain growth rate increased 2-fold in both strains; (ii) newly synthesized rRNA became unstable in both strains; (iii) the stable RNA gene activity (rRNA and tRNA, measured as stable RNA synthesis rate relative to the total instantaneous rate of RNA synthesis) decreased 1.7-fold in the relA+ strain and increased 1.9-fold in the relA mutant; and (iv) the RNA polymerase activity (measured by the percentage of total RNA polymerase enzyme active in transcription an any instant) decreased from 20 to 3.6% in the relA+ strain and remained unchanged (or increased at most to 22%) in the relA mutant. It is suggested that both rRNA gene activity and the RNA polymerase activity depend on the intracellular concentration of guanosine tetraphosphate, whereas the altered chain elongation rate and stability of rRNA are temperature or amino acid starvation effects, respectively, without involvement of relA function. PMID:6174501

  11. A Total Ozone Dependent Ozone Profile Climatology Based on Ozone-Sondes and Aura MLS Data

    NASA Astrophysics Data System (ADS)

    Labow, G. J.; McPeters, R. D.; Ziemke, J. R.

    2014-12-01

    A new total ozone-based ozone profile climatology has been created for use in satellite and/or ground based ozone retrievals. This climatology was formed by combining data from the Microwave Limb Sounder (MLS) with data from balloon sondes and binned by zone and total ozone. Because profile shape varies with total column ozone, this climatology better captures the ozone variations than the previously used seasonal climatologies, especially near the tropopause. This is significantly different than ozone climatologies used in the past as there is no time component. The MLS instrument on Aura has excellent latitude coverage and measures ozone profiles daily from the upper troposphere to the lower mesosphere at ~3.5 km resolution. Almost a million individual MLS ozone measurements are merged with data from over 55,000 ozonesondes which are then binned as a function of total ozone. The climatology consists of average ozone profiles as a function of total ozone for six 30 degree latitude bands covering altitudes from 0-75 km (in Z* pressure altitude coordinates). This new climatology better represents the profile shape as a function of total ozone than previous climatologies and shows some remarkable and somewhat unexpected correlations between total ozone and ozone in the lower altitudes, particularly in the lower and middle troposphere. These data can also be used to infer biases and errors in either the MLS retrievals or ozone sondes.

  12. Future body mass index modelling based on macronutrient profiles and physical activity

    PubMed Central

    2012-01-01

    Background An accurate system of determining the relationship of macronutrient profiles of foods and beverages to the long-term weight impacts of foods is necessary for evidence-based, unbiased front-of-the-package food labels. Methods Data sets on diet, physical activity, and BMI came from the Food and Agriculture Organization (FAO), the World Health Organization (WHO), the Diabetes Control and Complications Trial (DCCT), and Epidemiology Diabetes Intervention and Complications (EDIC). To predict future BMI of individuals, multiple regression derived FAO/WHO and DCCT/EDIC formulas related macronutrient profiles and physical activity (independent variables) to BMI change/year (dependent variable). Similar formulas without physical activity related macronutrient profiles of individual foods and beverages to four-year weight impacts of those items and compared those forecasts to published food group profiling estimates from three large prospective studies by Harvard nutritional epidemiologists. Results FAO/WHO food and beverage formula: four-year weight impact (pounds)=(0.07710 alcohol g+11.95 (381.7+carbohydrates g per serving)*4/(2,613+kilocalories per serving)–304.9 (30.38+dietary fiber g per serving)/(2,613+kilocalories per serving)+19.73 (84.44+total fat g)*9/(2,613+kilocalories per serving)–68.57 (20.45+PUFA g per serving)*9/(2,613+kilocalories per serving))*2.941–12.78 (n=334, R2=0.29, P < 0.0001). DCCT/EDIC formula for four-year weight impact (pounds)=(0.898 (102.2+protein g per serving)*4/(2,297+kilocalories per serving)+1.063 (264.2+carbohydrates g per serving)*4/(2,297+ kilocalories per serving)–13.19 (24.29+dietary fiber g per serving)/ (2,297+kilocalories per serving)+ 0.973 (74.59+(total fat g per serving–PUFA g per serving)*9/(2,297+kilocalories per serving))*85.82–68.11 (n=1,055, R2=0.03, P < 0.0001). (FAO/WHO+ DCCT/EDIC formula forecasts averaged correlated strongly with published food group profiling findings except for potatoes and

  13. BH3 profiling and a toolkit of BH3-mimetic drugs predict anti-apoptotic dependence of cancer cells

    PubMed Central

    Butterworth, Michael; Pettitt, Andrew; Varadarajan, Shankar; Cohen, Gerald M

    2016-01-01

    Background: Anti-apoptotic BCL-2 family members antagonise apoptosis by sequestering their pro-apoptotic counterparts. The balance between the different BCL-2 family members forms the basis of BH3 profiling, a peptide-based technique used to predict chemosensitivity of cancer cells. Recent identification of cell-permeable, selective inhibitors of BCL-2, BCL-XL and MCL-1, further facilitates the determination of the BCL-2 family dependency of cancer cells. Methods: We use BH3 profiling in combination with cell death analyses using a chemical inhibitor toolkit to assess chemosensitivity of cancer cells. Results: Both BH3 profiling and the inhibitor toolkit effectively predict chemosensitivity of cells addicted to a single anti-apoptotic protein but a combination of both techniques is more instructive when cell survival depends on more than one anti-apoptotic protein. Conclusions: The inhibitor toolkit provides a rapid, inexpensive and simple means to assess the chemosensitivity of tumour cells and in conjunction with BH3 profiling offers much potential in personalising cancer therapy. PMID:26954718

  14. Metabolomic Profiling of Prostate Cancer Progression During Active Surveillance

    DTIC Science & Technology

    2012-10-01

    that the profile captures the phenotype of Gleason grade 4 we will use clinically significant tumors from men who underwent immediate surgery (i.e. not...matched tumor and benign tissue samples from 5 open prostatectomy (RRP) cases and 5 robotic assisted laparoscopic prostatectomy (RAL) cases (including 3...criteria, collection of sufficient frozen tissue from the prostatectomy, matched to serum and urine collected prior to surgery , and collection of clinical

  15. Temperament and character model of personality profile of alcohol- and drug-dependent inpatients.

    PubMed

    Evren, Cuneyt; Evren, Bilge; Yancar, Cenk; Erkiran, Murat

    2007-01-01

    The aims of this study were to evaluate the differences in dimensions of temperament and character in Turkish alcohol- and drug-dependent inpatients, and to examine which dimensions would predict drug dependency. The subjects consisted of 111 alcohol-dependent and 93 drug-dependent inpatients according to the Diagnostic and Statistical Manual of Mental Disorders, 4th Edition. Subjects were tested using Cloninger's Temperament and Character Inventory (TCI). Among the temperament dimensions, novelty seeking score was higher and reward dependency score was lower in drug-dependent patients than in alcohol-dependent patients. Among the character dimensions, self-directedness and cooperativeness scores were lower in drug-dependent patients. Low age and novelty seeking predicted drug dependency in forward logistic regression model. Subscales that predicted drug dependency other than young age were lower scores on compassion vs revengefulness (C4) and helpfulness (C3), and higher score on spiritual acceptance vs rational materialism (ST3). As in previous studies, which indicate an association between personality and substance choice, in the present study, TCI was shown to be an efficient tool in discriminating alcohol and drug dependents; thus, it seems to be important to consider TCI dimensions in planning the treatment of substance dependency.

  16. Venus Then and Now: Simulating Sulfuric Acid Clouds Using Latitudinally Dependent VIRA and VeRA Temperature Profiles

    NASA Astrophysics Data System (ADS)

    Gao, P.; Parkinson, C. D.; Bardeen, C.; Yung, Y. L.

    2014-12-01

    Observations from the Pioneer Venus Orbiter (PVO) and from SPICAV/SOIR aboard Venus Express (VEx) have shown the upper haze (UH) of Venus to be highly spatially and temporally variable. Previous models of this system, using typical temperature profiles representative of the Venus atmosphere as a whole, did not investigate the effects of temperature variations on the UH particle distributions. Parkinson et al. (2014, submitted) showed that the inclusion of latitudinally dependent temperature profiles retrieved from SPICAV/SOIR observations in the Venus cloud model of Gao et al. (2014) resulted in markedly different cloud distributions between the different latitude cases, such as a lowered cloud base near the equator and a slightly thicker UH at the poles. Thus, temperature variations across Venus could help explain spatial variations in the atmospheric aerosol distribution. In this work, we expand on the aforementioned study by including VIRA temperature profiles derived from Venera and PVO observations (Kliore et al. 1985) at similar latitudes as the SPICAV/SOIR profiles to assess how the aerosol distribution varies spatially and temporally. By comparing the simulated cloud and haze distributions arising from the two sets of temperature profiles, we can evaluate whether secular changes have occurred in the ~30 years between the PVO and VEx epochs.

  17. Profile of male-field dependent (FD) prospective teacher's reflective thinking in solving contextual mathematical problem

    NASA Astrophysics Data System (ADS)

    Agustan, S.; Juniati, Dwi; Siswono, Tatag Yuli Eko

    2017-08-01

    Reflective thinking is an important component in the world of education, especially in professional education of teachers. In learning mathematics, reflective thinking is one way to solve mathematical problem because it can improve student's curiosity when student faces a mathematical problem. Reflective thinking is also a future competence that should be taught to students to face the challenges and to respond of demands of the 21st century. There are many factors which give impact toward the student's reflective thinking when student solves mathematical problem. One of them is cognitive style. For this reason, reflective thinking and cognitive style are important things in solving contextual mathematical problem. This research paper describes aspect of reflective thinking in solving contextual mathematical problem involved solution by using some mathematical concept, namely linear program, algebra arithmetic operation, and linear equations of two variables. The participant, in this research paper, is a male-prospective teacher who has Field Dependent. The purpose of this paper is to describe aspect of prospective teachers' reflective thinking in solving contextual mathematical problem. This research paper is a descriptive by using qualitative approach. To analyze the data, the researchers focus in four main categories which describe prospective teacher's activities using reflective thinking, namely; (a) formulation and synthesis of experience, (b) orderliness of experience, (c) evaluating the experience and (d) testing the selected solution based on the experience.

  18. Solutions of the Bogoliubov-de Gennes equation with position dependent Fermi-velocity and gap profiles

    NASA Astrophysics Data System (ADS)

    Presilla, M.; Panella, O.; Roy, P.

    2017-02-01

    It is shown that bound state solutions of the one dimensional Bogoliubov-de Gennes (BdG) equation may exist when the Fermi velocity becomes dependent on the space coordinate. The existence of bound states in continuum (BIC) like solutions has also been confirmed both in the normal phase as well as in the super-conducting phase. We also show that a combination of Fermi velocity and gap parameter step-like profiles provides scattering solutions with normal reflection and transmission.

  19. Activity-Dependent Neuronal Model on Complex Networks

    PubMed Central

    de Arcangelis, Lucilla; Herrmann, Hans J.

    2012-01-01

    Neuronal avalanches are a novel mode of activity in neuronal networks, experimentally found in vitro and in vivo, and exhibit a robust critical behavior: these avalanches are characterized by a power law distribution for the size and duration, features found in other problems in the context of the physics of complex systems. We present a recent model inspired in self-organized criticality, which consists of an electrical network with threshold firing, refractory period, and activity-dependent synaptic plasticity. The model reproduces the critical behavior of the distribution of avalanche sizes and durations measured experimentally. Moreover, the power spectra of the electrical signal reproduce very robustly the power law behavior found in human electroencephalogram (EEG) spectra. We implement this model on a variety of complex networks, i.e., regular, small-world, and scale-free and verify the robustness of the critical behavior. PMID:22470347

  20. Effects of activation energy and activation volume on the temperature-dependent viscosity of water.

    PubMed

    Kwang-Hua, Chu Rainer

    2016-08-01

    Water transport in a leaf is vulnerable to viscosity-induced changes. Recent research has suggested that these changes may be partially due to variation at the molecular scale, e.g., regulations via aquaporins, that induce reductions in leaf hydraulic conductance. What are the quantitative as well as qualitative changes in temperature-dependent viscosity due to the role of aquaporins in tuning activation energy and activation volume? Using the transition-state approach as well as the boundary perturbation method, we investigate temperature-dependent viscosity tuned by activation energy and activation volume. To validate our approach, we compare our numerical results with previous temperature-dependent viscosity measurements. The rather good fit between our calculations and measurements confirms our present approach. We have obtained critical parameters for the temperature-dependent (shear) viscosity of water that might be relevant to the increasing and reducing of leaf hydraulic conductance. These parameters are sensitive to temperature, activation energy, and activation volume. Once the activation energy increases, the (shear) viscosity of water increases. Our results also show that as the activation volume increases (say, 10^{-23}m^{3}), the (shear) viscosity of water decreases significantly and the latter induces the enhancing of leaf hydraulic conductance. Within the room-temperature regime, a small increase in the activation energy will increase the water viscosity or reduce the leaf hydraulic conductance. Our approach and results can be applied to diverse plant or leaf attributes.

  1. Effects of activation energy and activation volume on the temperature-dependent viscosity of water

    NASA Astrophysics Data System (ADS)

    Kwang-Hua, Chu Rainer

    2016-08-01

    Water transport in a leaf is vulnerable to viscosity-induced changes. Recent research has suggested that these changes may be partially due to variation at the molecular scale, e.g., regulations via aquaporins, that induce reductions in leaf hydraulic conductance. What are the quantitative as well as qualitative changes in temperature-dependent viscosity due to the role of aquaporins in tuning activation energy and activation volume? Using the transition-state approach as well as the boundary perturbation method, we investigate temperature-dependent viscosity tuned by activation energy and activation volume. To validate our approach, we compare our numerical results with previous temperature-dependent viscosity measurements. The rather good fit between our calculations and measurements confirms our present approach. We have obtained critical parameters for the temperature-dependent (shear) viscosity of water that might be relevant to the increasing and reducing of leaf hydraulic conductance. These parameters are sensitive to temperature, activation energy, and activation volume. Once the activation energy increases, the (shear) viscosity of water increases. Our results also show that as the activation volume increases (say, 10-23m3 ), the (shear) viscosity of water decreases significantly and the latter induces the enhancing of leaf hydraulic conductance. Within the room-temperature regime, a small increase in the activation energy will increase the water viscosity or reduce the leaf hydraulic conductance. Our approach and results can be applied to diverse plant or leaf attributes.

  2. Protein Profiling and Histone Deacetylation Activities in Somaclonal Variants of Oil Palm (Elaeis guineensis Jacq.)

    PubMed Central

    Yaacob, Jamilah Syafawati; Loh, Hwei-San; Mat Taha, Rosna

    2013-01-01

    Mantled fruits as a result of somaclonal variation are often observed from the oil palm plantlets regenerated via tissue culture. The mantling of fruits with finger-like and thick outer coating phenotypes significantly reduces the seed size and oil content, posing a threat to oil palm planters, and may jeopardize the economic growth of countries that depend particularly on oil palm plantation. The molecular aspects of the occurrence of somaclonal variations are yet to be known, possibly due to gene repression such as DNA methylation, histone methylation and histone deacetylation. Histone deacetylases (HDACs), involved in eukaryotic gene regulation by catalyzing the acetyl groups are removal from lysine residues on histone, hence transcriptionally repress gene expression. This paper described the total protein polymorphism profiles of somaclonal variants of oil palm and the effects of histone deacetylation on this phenomenon. Parallel to the different phenotypes, the protein polymorphism profiles of the mantled samples (leaves, fruits, and florets) and the phenotypically normal samples were proven to be different. Higher HDAC activity was found in mantled leaf samples than in the phenotypically normal leaf samples, leading to a preliminary conclusion that histone deacetylation suppressed gene expression and contributed to the development of somaclonal variants. PMID:23844406

  3. Protein profiling and histone deacetylation activities in somaclonal variants of oil palm (Elaeis guineensis Jacq.).

    PubMed

    Yaacob, Jamilah Syafawati; Loh, Hwei-San; Mat Taha, Rosna

    2013-01-01

    Mantled fruits as a result of somaclonal variation are often observed from the oil palm plantlets regenerated via tissue culture. The mantling of fruits with finger-like and thick outer coating phenotypes significantly reduces the seed size and oil content, posing a threat to oil palm planters, and may jeopardize the economic growth of countries that depend particularly on oil palm plantation. The molecular aspects of the occurrence of somaclonal variations are yet to be known, possibly due to gene repression such as DNA methylation, histone methylation and histone deacetylation. Histone deacetylases (HDACs), involved in eukaryotic gene regulation by catalyzing the acetyl groups are removal from lysine residues on histone, hence transcriptionally repress gene expression. This paper described the total protein polymorphism profiles of somaclonal variants of oil palm and the effects of histone deacetylation on this phenomenon. Parallel to the different phenotypes, the protein polymorphism profiles of the mantled samples (leaves, fruits, and florets) and the phenotypically normal samples were proven to be different. Higher HDAC activity was found in mantled leaf samples than in the phenotypically normal leaf samples, leading to a preliminary conclusion that histone deacetylation suppressed gene expression and contributed to the development of somaclonal variants.

  4. SU-E-T-299: Small Fields Profiles Correction Through Detectors Spatial Response Functions and Field Size Dependence Analysis

    SciTech Connect

    Filipuzzi, M; Garrigo, E; Venencia, C; Germanier, A

    2014-06-01

    Purpose: To calculate the spatial response function of various radiation detectors, to evaluate the dependence on the field size and to analyze the small fields profiles corrections by deconvolution techniques. Methods: Crossline profiles were measured on a Novalis Tx 6MV beam with a HDMLC. The configuration setup was SSD=100cm and depth=5cm. Five fields were studied (200×200mm2,100×100mm2, 20×20mm2, 10×10mm2and 5×5mm2) and measured were made with passive detectors (EBT3 radiochromic films and TLD700 thermoluminescent detectors), ionization chambers (PTW30013, PTW31003, CC04 and PTW31016) and diodes (PTW60012 and IBA SFD). The results of passive detectors were adopted as the actual beam profile. To calculate the detectors kernels, modeled by Gaussian functions, an iterative process based on a least squares criterion was used. The deconvolutions of the measured profiles were calculated with the Richardson-Lucy method. Results: The profiles of the passive detectors corresponded with a difference in the penumbra less than 0.1mm. Both diodes resolve the profiles with an overestimation of the penumbra smaller than 0.2mm. For the other detectors, response functions were calculated and resulted in Gaussian functions with a standard deviation approximate to the radius of the detector in study (with a variation less than 3%). The corrected profiles resolve the penumbra with less than 1% error. Major discrepancies were observed for cases in extreme conditions (PTW31003 and 5×5mm2 field size). Conclusion: This work concludes that the response function of a radiation detector is independent on the field size, even for small radiation beams. The profiles correction, using deconvolution techniques and response functions of standard deviation equal to the radius of the detector, gives penumbra values with less than 1% difference to the real profile. The implementation of this technique allows estimating the real profile, freeing from the effects of the detector used for the

  5. Degrasyn activates proteasomal-dependent degradation of c-Myc.

    PubMed

    Bartholomeusz, Geoffrey; Talpaz, Moshe; Bornmann, William; Kong, Ling-Yuan; Donato, Nicholas J

    2007-04-15

    c-Myc is a highly unstable transcription factor whose deregulation and increased expression are associated with cancer. Degrasyn, a small synthetic molecule, induces rapid degradation of c-Myc protein in MM-1 multiple myeloma and other tumor cell lines. Destruction of c-Myc by degrasyn requires the presence of a region of c-Myc between amino acid residues 316 and 378 that has not previously been associated with c-Myc stability. Degrasyn-induced degradation of c-Myc depends on proteasomes but is independent of the degron regions previously shown to be important for ubiquitin-mediated targeting and proteasomal destruction of the protein. Degrasyn-dependent c-Myc proteolysis is not mediated by any previously identified c-Myc regulatory mechanism, does not require new protein synthesis, and does not depend on the nuclear localization of c-Myc. Degrasyn reduced c-Myc levels in A375 melanoma cells and in A375 tumors in nude mice, and this activity correlated with tumor growth inhibition. Together, these results suggest that degrasyn reduces the stability of c-Myc in vitro and in vivo through a unique signaling process that uses c-Myc domains not previously associated with c-Myc regulation.

  6. Parthanatos Mediates AIMP2 Activated Age Dependent Dopaminergic Neuronal Loss

    PubMed Central

    Lee, Yunjong; Karuppagounder, Senthilkumar S.; Shin, Joo-Ho; Lee, Yun-Il; Ko, Han Seok; Swing, Debbie; Jiang, Haisong; Kang, Sung-Ung; Lee, Byoung Dae; Kang, Ho Chul; Kim, Donghoon; Tessarollo, Lino; Dawson, Valina L.; Dawson, Ted M.

    2013-01-01

    The defining pathogenic feature of Parkinson’s disease is the age dependent loss of dopaminergic neurons. Mutations and inactivation of parkin, an ubiquitin E3 ligase, cause Parkinson’s disease through accumulation of pathogenic substrates. Here we show that transgenic overexpression of the parkin substrate, aminoacyl-tRNA synthetase complex interacting multifunctional protein-2 (AIMP2) leads to a selective, age-dependent progressive loss of dopaminergic neurons via activation of poly(ADP-ribose) polymerase-1 (PARP1). AIMP2 accumulation in vitro and in vivo results in PARP1 overactivation and dopaminergic cell toxicity via direct association of these proteins in the nucleus providing a new path to PARP1 activation other than DNA damage. Inhibition of PARP1 through gene deletion or drug inhibition reverses behavioral deficits and protects in vivo against dopamine neuron death in AIMP2 transgenic mice. These data indicate that brain permeable PARP inhibitors could be effective in delaying or preventing disease progression in Parkinson’s disease. PMID:23974709

  7. Light-dependent activation of rod transducin by pineal opsin.

    PubMed

    Max, M; Surya, A; Takahashi, J S; Margolskee, R F; Knox, B E

    1998-10-09

    The pineal gland expresses a unique member of the opsin family (P-opsin; Max, M., McKinnon, P. J., Seidenman, K. J., Barrett, R. K., Applebury, M. L., Takahashi, J. S., and Margolskee, R. F. (1995) Science 267, 1502-1506) that may play a role in circadian entrainment and photo-regulation of melatonin synthesis. To study the function of this protein, an epitope-tagged P-opsin was stably expressed in an embryonic chicken pineal cell line. When incubated with 11-cis-retinal, a light-sensitive pigment was formed with a lambdamax at 462 +/- 2 nm. P-opsin bleached slowly in the dark (t1/2 = 2 h) in the presence of 50 mM hydroxylamine. Purified P-opsin in dodecyl maltoside activated rod transducin in a light-dependent manner, catalyzing the exchange of more than 300 mol of GTPgammaS (guanosine 5'-O-(3-thiotriphosphate))/mol of P-opsin. The initial rate for activation (75 mol of GTPgammaS bound/mol of P-opsin/min at 7 microM) increased with increasing concentrations of transducin. The addition of egg phosphatidylcholine to P-opsin had little effect on the activation kinetics; however, the intrinsic rate of decay in the absence of transducin was accelerated. These results demonstrate that P-opsin is an efficient catalyst for activation of rod transducin and suggest that the pineal gland may contain a rodlike phototransduction cascade.

  8. Temporal dependence of cysteine protease activation following excitotoxic hippocampal injury.

    PubMed

    Berry, J N; Sharrett-Field, L J; Butler, T R; Prendergast, M A

    2012-10-11

    Excitotoxic insults can lead to intracellular signaling cascades that contribute to cell death, in part by activation of proteases, phospholipases, and endonucleases. Cysteine proteases, such as calpains, are calcium (Ca(2+))-activated enzymes which degrade cytoskeletal proteins, including microtubule-associated proteins, tubulin, and spectrin, among others. The current study used the organotypic hippocampal slice culture model to examine whether pharmacologic inhibition of cysteine protease activity inhibits N-methyl-D-aspartate- (NMDA-) induced excitotoxic (20 μM NMDA) cell death and changes in synaptophysin immunoreactivity. Significant NMDA-induced cytotoxicity (as measured by propidium iodide [PI] uptake) was found in the CA1 region of the hippocampus at all timepoints examined (24, 72, 120 h), an effect significantly attenuated by co-exposure to the selective NMDA receptor antagonist DL-2-Amino-5-phosphonopentanoic acid (APV), but not MDL-28170, a potent cysteine protease inhibitor. Results indicated sparing of NMDA-induced loss of the synaptic vesicular protein synaptophysin in all regions of the hippocampus by MDL-28170, though only at early timepoints after injury. These results suggest Ca(2+)-dependent recruitment of cysteine proteases within 24h of excitotoxic insult, but activation of alternative cellular degrading mechanisms after 24h. Further, these data suggest that synaptophysin may be a substrate for calpains and related proteases.

  9. Chk2 Activation and Phosphorylation-Dependent Oligomerization

    PubMed Central

    Xu, Xingzhi; Tsvetkov, Lyuben M.; Stern, David F.

    2002-01-01

    The tumor suppressor gene CHK2 encodes a versatile effector serine/threonine kinase involved in responses to DNA damage. Chk2 has an amino-terminal SQ/TQ cluster domain (SCD), followed by a forkhead-associated (FHA) domain and a carboxyl-terminal kinase catalytic domain. Mutations in the SCD or FHA domain impair Chk2 checkpoint function. We show here that autophosphorylation of Chk2 produced in a cell-free system requires trans phosphorylation by a wortmannin-sensitive kinase, probably ATM or ATR. Both SQ/TQ sites and non-SQ/TQ sites within the Chk2 SCD can be phosphorylated by active Chk2. Amino acid substitutions in the SCD and the FHA domain impair auto- and trans-kinase activities of Chk2. Chk2 forms oligomers that minimally require the FHA domain of one Chk2 molecule and the SCD within another Chk2 molecule. Chk2 oligomerization in vivo increases after DNA damage, and when damage is induced by gamma irradiation, this increase requires ATM. Chk2 oligomerization is phosphorylation dependent and can occur in the absence of other eukaryotic proteins. Chk2 can cross-phosphorylate another Chk2 molecule in an oligomeric complex. Induced oligomerization of a Chk2 chimera in vivo concomitant with limited DNA damage augments Chk2 kinase activity. These results suggest that Chk2 oligomerization regulates Chk2 activation, signal amplification, and transduction in DNA damage checkpoint pathways. PMID:12024051

  10. Task-dependent posterior cingulate activation in mild cognitive impairment.

    PubMed

    Ries, Michele L; Schmitz, Taylor W; Kawahara, Tisha N; Torgerson, Britta M; Trivedi, Mehul A; Johnson, Sterling C

    2006-01-15

    Neuroimaging research has demonstrated that the posterior cingulate cortex (PCC) is functionally compromised in individuals diagnosed with amnestic mild cognitive impairment (MCI), a major risk factor for the development of Alzheimer's disease (AD). In functional MRI studies with healthy participants, this same region is active during self-appraisal (requiring retrieval of semantic knowledge about the self) as well as episodic recognition of previously learned information. Administering both types of tasks to people with MCI may reveal important information on the role of the PCC in recollection. This study investigated fMRI activation in the PCC in individuals with MCI and matched controls across two tasks. The first task was a visual episodic recognition task. The second task was an autobiographical self-appraisal task in which subjects rated themselves on a set of trait adjectives. Results of a conjunction analysis revealed the PCC as the sole region commonly active during both tasks in the healthy older adults. Furthermore, additional analysis revealed an interaction in the PCC, indicating a task-dependent response in the MCI group. MCI participants showed PCC activation during self-appraisal, but not episodic retrieval. This result suggests in MCI that the PCC shows functional degradation during episodic retrieval; however, the PCC's role in retrieval and evaluation of highly elaborated information regarding the self is more well-preserved.

  11. Time-dependent profiling of metabolites from Snf1 mutant and wild type yeast cells.

    PubMed

    Humston, Elizabeth M; Dombek, Kenneth M; Hoggard, Jamin C; Young, Elton T; Synovec, Robert E

    2008-11-01

    The effect of sampling time in the context of growth conditions on a dynamic metabolic system was investigated in order to assess to what extent a single sampling time may be sufficient for general application, as well as to determine if useful kinetic information could be obtained. A wild type yeast strain (W) was compared to a snf1Delta mutant yeast strain (S) grown in high-glucose medium (R) and in low-glucose medium containing ethanol (DR). Under these growth conditions, different metabolic pathways for utilizing the different carbon sources are expected to be active. Thus, changes in metabolite levels relating to the carbon source in the growth medium were anticipated. Furthermore, the Snf1 protein kinase complex is required to adapt cellular metabolism from fermentative R conditions to oxidative DR conditions. So, differences in intracellular metabolite levels between the W and S yeast strains were also anticipated. Cell extracts were collected at four time points (0.5, 2, 4, 6 h) after shifting half of the cells from R to DR conditions, resulting in 16 sample classes (WR, WDR, SR, SDR) x (0.5, 2, 4, 6 h). The experimental design provided time course data, so temporal dependencies could be monitored in addition to carbon source and strain dependencies. Comprehensive two-dimensional (2D) gas chromatography coupled to time-of-flight mass spectrometry (GC x GC-TOFMS) was used with discovery-based data mining algorithms ( Anal. Chem. 2006, 78, 5068-5075 (ref 1); J. Chromatogr., A 2008, 1186, 401-411 (ref 2)) to locate regions within the 2D chromatograms (i.e., metabolites) that provided chemical selectivity between the 16 sample classes. These regions were mathematically resolved using parallel factor analysis to positively identify the metabolites and to acquire quantitative results. With these tools, 51 unique metabolites were identified and quantified. Various time course patterns emerged from these data, and principal component analysis (PCA) was utilized as

  12. Quantitative Time Profiling of Children's Activity and Motion.

    PubMed

    Barnes, Claire M; Clark, Cain C T; Holton, Mark D; Stratton, Gareth; Summers, Huw D

    2017-01-01

    The aim of this study was to establish children's mechanical movement patterns during a standardized assessment of fitness by using an accelerometer. Further to this, our objective was to use the information from the accelerometer to profile individual time courses of exercise, across the cohort. A multistage fitness test study was performed with 103 children (mean ± SD age = 10.3 ± 0.6 yr). Children wore an ankle-mounted accelerometer, and gait data were collected on radial acceleration traces obtained at a frequency of 40 Hz. Time-resolved metrics of foot impact force, maximum leg lift angle, and stride frequency were used to profile children's performance across the test duration. A whole-history metric of stride quality, based on the changing ratio of stride length to stride frequency, was used in bivariate analyses of physical performance and body metrics. Stride angle derived by our protocol was found to have a strong positive correlation with integrated acceleration, synonymous with counts, widely used in the sport science community (r = 0.81, 0.79, and 0.80 across different stages of the multistage fitness test). Accelerometer data show that differing performance in the test is related to the children's ability to accurately control their gait, with high performers displaying a linearly increasing speed, delivered through stride extension and well matched to the demand level of the test. A negative correlation was found between stride quality and body measures of body mass index (r = -0.61) and body mass (r = -0.60). Profiles of the gait parameters provide information on the mechanics of child's motion, allowing detailed assessment of multiple parameter during increasing intensities of exercise.

  13. Activity-Based Protein Profiling of Ammonia Monooxygenase in Nitrosomonas europaea

    PubMed Central

    Bennett, Kristen; Sadler, Natalie C.; Wright, Aaron T.; Yeager, Chris

    2016-01-01

    Nitrosomonas europaea is an aerobic nitrifying bacterium that oxidizes ammonia (NH3) to nitrite (NO2−) through the sequential activities of ammonia monooxygenase (AMO) and hydroxylamine dehydrogenase (HAO). Many alkynes are mechanism-based inactivators of AMO, and here we describe an activity-based protein profiling method for this enzyme using 1,7-octadiyne (17OD) as a probe. Inactivation of NH4+-dependent O2 uptake by N. europaea by 17OD was time- and concentration-dependent. The effects of 17OD were specific for ammonia-oxidizing activity, and de novo protein synthesis was required to reestablish this activity after cells were exposed to 17OD. Cells were reacted with Alexa Fluor 647 azide using a copper-catalyzed azide-alkyne cycloaddition (CuAAC) (click) reaction, solubilized, and analyzed by SDS-PAGE and infrared (IR) scanning. A fluorescent 28-kDa polypeptide was observed for cells previously exposed to 17OD but not for cells treated with either allylthiourea or acetylene prior to exposure to 17OD or for cells not previously exposed to 17OD. The fluorescent polypeptide was membrane associated and aggregated when heated with β-mercaptoethanol and SDS. The fluorescent polypeptide was also detected in cells pretreated with other diynes, but not in cells pretreated with structural homologs containing a single ethynyl functional group. The membrane fraction from 17OD-treated cells was conjugated with biotin-azide and solubilized in SDS. Streptavidin affinity-purified polypeptides were on-bead trypsin-digested, and amino acid sequences of the peptide fragments were determined by liquid chromatography-mass spectrometry (LC-MS) analysis. Peptide fragments from AmoA were the predominant peptides detected in 17OD-treated samples. In-gel digestion and matrix-assisted laser desorption ionization–tandem time of flight (MALDI-TOF/TOF) analyses also confirmed that the fluorescent 28-kDa polypeptide was AmoA. PMID:26826234

  14. Disaster Preparedness Among Active Duty Navy, Retirees, Veterans, and Dependents

    PubMed Central

    Annis, Heather; Jacoby, Irving; DeMers, Gerard

    2016-01-01

    Background With the increase in natural and manmade disasters, preparedness remains a vital area of concern. Despite attempts by government and non-government agencies to stress the importance of preparedness, national levels of preparedness remain unacceptably low. A goal of commands and installations is to ensure that US Navy beneficiaries are well prepared for disasters. This is especially critical in active service members to meet mission readiness requirements in crisis settings. Objective To evaluate active duty personnel, dependents, veterans, and retirees regarding disaster preparedness status. Methods The authors conducted an anonymous 29-question survey for US Navy active duty, dependents, veterans and retirees of the Greater San Diego Region evaluating actual basic disaster readiness as determined by FEMA standards of 3-day minimum supply of emergency stores and equipment. Descriptive statistics and regression analysis were used to analyze data. Results 1150 surveys were returned and analyzed. 983 were sufficiently complete for logistic regression analysis with 394 responding “Yes” to having a 72-hour disaster kit (40.1%) while 589 had “No” as a response (59.9%). Conclusion The surveyed population is no more prepared than the general public though surveyed beneficiaries overall are at upper range of preparedness. Lower income and levels of education were associated with lack of preparedness, whereas training in disaster preparedness or having been affected by disasters increased the likelihood of being adequately prepared. Unlike results seen in the general public, those with chronic healthcare needs in our surveyed population were more, rather than less likely to be prepared and those with minor children were less likely, rather than more likely to be prepared. Duty status was assessed and only veterans were significantly more likely than most to be prepared. PMID:26903142

  15. Disaster Preparedness among Active Duty Personnel, Retirees, Veterans, and Dependents.

    PubMed

    Annis, Heather; Jacoby, Irving; DeMers, Gerard

    2016-04-01

    With the increase in natural and manmade disasters, preparedness remains a vital area of concern. Despite attempts by government and non-government agencies to stress the importance of preparedness, national levels of preparedness remain unacceptably low. A goal of commands and installations is to ensure that US Navy beneficiaries are well prepared for disasters. This especially is critical in active service members to meet mission readiness requirements in crisis settings. To evaluate active duty Navy personnel, dependents, veterans, and retirees regarding disaster preparedness status. The authors conducted an anonymous 29-question survey for US Navy active duty, dependents, veterans, and retirees of the Greater San Diego Region (California, USA) evaluating actual basic disaster readiness as determined by the Federal Emergency Management Agency (FEMA) standards of 3-day minimum supply of emergency stores and equipment. Descriptive statistics and regression analysis were used to analyze data. One thousand one hundred and fifty surveys were returned and analyzed. Nine hundred and eight-three were sufficiently complete for logistic regression analysis with 394 responding "Yes" to having a 72-hour disaster kit (40.1%) while 589 had "No" as a response (59.9%). The surveyed population is no more prepared than the general public, though surveyed beneficiaries overall are at an upper range of preparedness. Lower income and levels of education were associated with lack of preparedness, whereas training in disaster preparedness or having been affected by disasters increased the likelihood of being adequately prepared. Unlike results seen in the general public, those with chronic health care needs in the surveyed population were more, rather than less, likely to be prepared and those with minor children were less likely, rather than more likely, to be prepared. Duty status was assessed and only veterans were emphatically more probable than most to be prepared.

  16. Assessing Sources of Stress to Aquatic Ecosystems: Using Biomarkers and Bioindicators to Characterize Exodure-Response Profiles of Anthropogenic Activities

    SciTech Connect

    Adams, S.M.

    1999-03-29

    Establishing causal relationships between sources of environmental stressors and aquatic ecosystem health if difficult because of the many biotic and abiotic factors which can influence or modify responses of biological systems to stress, the orders of magnitude involved in extrapolation over both spatial and temporal scales, and compensatory mechanisms such as density-dependent responses that operate in populations. To address the problem of establishing causality between stressors and effects on aquatic systems, a diagnostic approach, based on exposure-response profiles for various anthropogenic activities, was developed to help identify sources of stress responsible for effects on aquatic systems at ecological significant levels of biological organization (individual, population, community). To generate these exposure-effects profiles, biomarkers of exposure were plotted against bioindicators of corresponding effects for several major anthropogenic activities including petrochemical , pulp and paper, domestic sewage, mining operations, land-development activities, and agricultural activities. Biomarkers of exposure to environmental stressors varied depending on the type of anthropogenic activity involved. Bioindicator effects, however, including histopathological lesions, bioenergetic status, individual growth, reproductive impairment, and community-level responses were similar among many of the major anthropogenic activities. This approach is valuable to help identify and diagnose sources of stressors in environments impacted by multiple stressors. By identifying the types and sources of environmental stressors, aquatic ecosystems can be more effectively protected and managed to maintain acceptable levels of environmental quality and ecosystem fitness.

  17. The Relation between Profiles of Leisure Activity Participation and Substance Use among South African Youth.

    PubMed

    Tibbits, Melissa K; Caldwell, Linda L; Smith, Edward A; Wegner, Lisa

    2009-01-01

    A large body of research indicates that leisure activity participation is associated with substance use for American adolescents, and that leisure may be an important context of substance use prevention. It is important to begin to apply what we have learned with American youth and extend the knowledge base in other countries with significant adolescent risk behavior. The current study examined the association between leisure activity participation and substance use among a predominately Colored sample of 3497 South African 8(th) graders. Males' activity participation was characterized by five leisure activity profiles (Uninvolved; Sports and Volunteer; Mixed: Recreation and Hobbies; Mixed: Artistic; Highly Involved), whereas females' activity participation was characterized by four leisure activity profiles (Uninvolved; Uninvolved but Social; Mixed; Highly Involved). Leisure activity profiles were significantly associated with past-month alcohol, tobacco, and marijuana use.

  18. Dehydrogenase activity of forest soils depends on the assay used

    NASA Astrophysics Data System (ADS)

    Januszek, Kazimierz; Długa, Joanna; Socha, Jarosław

    2015-01-01

    Dehydrogenases are exclusively intracellular enzymes, which play an important role in the initial stages of oxidation of soil organic matter. One of the most frequently used methods to estimate dehydrogenase activity in soil is based on the use of triphenyltetrazolium chloride as an artificial electron acceptor. The purpose of this study was to compare the activity of dehydrogenases of forest soils with varied physicochemical properties using different triphenyltetrazolium chloride assays. The determination was carried out using the original procedure by Casida et al., a modification of the procedure which involves the use of Ca(OH)2 instead of CaCO3, the Thalmann method, and the assay by Casida et al. without addition of buffer or any salt. Soil dehydrogenase activity depended on the assay used. Dehydrogenase determined by the Casida et al. method without addition of buffer or any salt correlated with the pH values of soils. The autoclaved strongly acidic samples of control soils showed high concentrations of triphenylformazan, probably due to chemical reduction of triphenyltetrazolium chloride. There is, therefore, a need for a sterilization method other than autoclaving, ie a process that results in significant changes in soil properties, thus helping to increase the chemical reduction of triphenyltetrazolium chloride.

  19. Sex-dependent differences in voluntary physical activity.

    PubMed

    Rosenfeld, Cheryl S

    2017-01-02

    Numbers of overweight and obese individuals are increasing in the United States and globally, and, correspondingly, the associated health care costs are rising dramatically. More than one-third of children are currently considered obese with a predisposition to type 2 diabetes, and it is likely that their metabolic conditions will worsen with age. Physical inactivity has also risen to be the leading cause of many chronic, noncommunicable diseases (NCD). Children are more physically inactive now than they were in past decades, which may be due to intrinsic and extrinsic factors. In rodents, the amount of time engaged in spontaneous activity within the home cage is a strong predictor of later adiposity and weight gain. Thus, it is important to understand primary motivators stimulating physical activity (PA). There are normal sex differences in PA levels in rodents and humans. The perinatal environment can induce sex-dependent differences in PA disturbances. This Review considers the current evidence for sex differences in PA in rodents and humans. The rodent studies showing that early exposure to environmental chemicals can shape later adult PA responses are discussed. Next, whether there are different motivators stimulating exercise in male vs. female humans are examined. Finally, the brain regions, genes, and pathways that modulate PA in rodents, and possibly by translation in humans, are described. A better understanding of why each sex remains physically active through the life span could open new avenues for preventing and treating obesity in children and adults. © 2016 Wiley Periodicals, Inc.

  20. The DNA methylation profile of activated human natural killer cells.

    PubMed

    Wiencke, John K; Butler, Rondi; Hsuang, George; Eliot, Melissa; Kim, Stephanie; Sepulveda, Manuel A; Siegel, Derick; Houseman, E Andres; Kelsey, Karl T

    2016-05-03

    Natural killer (NK) cells are now recognized to exhibit characteristics akin to cells of the adaptive immune system. The generation of adaptive memory is linked to epigenetic reprogramming including alterations in DNA methylation. The study herein found reproducible genome wide DNA methylation changes associated with human NK cell activation. Activation led predominately to CpG hypomethylation (81% of significant loci). Bioinformatics analysis confirmed that non-coding and gene-associated differentially methylated sites (DMS) are enriched for immune related functions (i.e., immune cell activation). Known DNA methylation-regulated immune loci were also identified in activated NK cells (e.g., TNFA, LTA, IL13, CSF2). Twenty-one loci were designated high priority and further investigated as potential markers of NK activation. BHLHE40 was identified as a viable candidate for which a droplet digital PCR assay for demethylation was developed. The assay revealed high demethylation in activated NK cells and low demethylation in naïve NK, T- and B-cells. We conclude the NK cell methylome is plastic with potential for remodeling. The differentially methylated region signature of activated NKs revealed similarities with T cell activation, but also provided unique biomarker candidates of NK activation, which could be useful in epigenome-wide association studies to interrogate the role of NK subtypes in global methylation changes associated with exposures and/or disease states.

  1. The DNA methylation profile of activated human natural killer cells

    PubMed Central

    Wiencke, John. K.; Butler, Rondi; Hsuang, George; Eliot, Melissa; Kim, Stephanie; Sepulveda, Manuel A.; Siegel, Derick; Houseman, E. Andres; Kelsey, Karl T.

    2016-01-01

    ABSTRACT Natural killer (NK) cells are now recognized to exhibit characteristics akin to cells of the adaptive immune system. The generation of adaptive memory is linked to epigenetic reprogramming including alterations in DNA methylation. The study herein found reproducible genome wide DNA methylation changes associated with human NK cell activation. Activation led predominately to CpG hypomethylation (81% of significant loci). Bioinformatics analysis confirmed that non-coding and gene-associated differentially methylated sites (DMS) are enriched for immune related functions (i.e., immune cell activation). Known DNA methylation-regulated immune loci were also identified in activated NK cells (e.g., TNFA, LTA, IL13, CSF2). Twenty-one loci were designated high priority and further investigated as potential markers of NK activation. BHLHE40 was identified as a viable candidate for which a droplet digital PCR assay for demethylation was developed. The assay revealed high demethylation in activated NK cells and low demethylation in naïve NK, T- and B-cells. We conclude the NK cell methylome is plastic with potential for remodeling. The differentially methylated region signature of activated NKs revealed similarities with T cell activation, but also provided unique biomarker candidates of NK activation, which could be useful in epigenome-wide association studies to interrogate the role of NK subtypes in global methylation changes associated with exposures and/or disease states. PMID:26967308

  2. Tool use ability depends on understanding of functional dynamics and not specific joint contribution profiles

    PubMed Central

    Parry, Ross; Dietrich, Gilles; Bril, Blandine

    2014-01-01

    Researchers in cognitive neuroscience have become increasingly interested in how different aspects of tool use are integrated and represented by the brain. Comparatively less attention has been directed toward tool use actions themselves and how effective tool use behaviors are coordinated. In response, we take this opportunity to consider the mechanical principles of tool use actions and their relationship to motor learning. Using kinematic analysis, we examine both functional dynamics and joint contribution profiles of subjects with different levels of experience in a primordial percussive task. Our results show that the ability to successfully produce stone flakes using the Oldowan method did not correspond with any particular joint contribution profile. Rather, expertise in this tool use action was principally associated with the subject's ability to regulate the functional parameters that define the task itself. PMID:24795669

  3. Plant secondary metabolite profiling evidences strain-dependent effect in the Azospirillum-Oryza sativa association.

    PubMed

    Chamam, Amel; Sanguin, Hervé; Bellvert, Floriant; Meiffren, Guillaume; Comte, Gilles; Wisniewski-Dyé, Florence; Bertrand, Cédric; Prigent-Combaret, Claire

    2013-03-01

    Azospirillum is a plant growth-promoting rhizobacterium (PGPR) able to enhance growth and yield of cereals such as rice, maize and wheat. The growth-promoting ability of some Azospirillum strains appears to be highly specific to certain plant species and cultivars. In order to ascertain the specificity of the associative symbiosis between rice and Azospirillum, the physiological response of two rice cultivars, Nipponbare and Cigalon, inoculated with two rice-associated Azospirillum was analyzed at two levels: plant growth response and plant secondary metabolic response. Each strain of Azospirillum (Azospirillum lipoferum 4B isolated from Cigalon and Azospirillum sp. B510 isolated from Nipponbare) preferentially increased growth of the cultivar from which it was isolated. This specific effect is not related to a defect in colonization of host cultivar as each strain colonizes effectively both rice cultivars, either at the rhizoplane (for 4B and B510) and inside the roots (for B510). The metabolic profiling approach showed that, in response to PGPR inoculation, profiles of rice secondary metabolites were modified, with phenolic compounds such as flavonoids and hydroxycinnamic derivatives being the main metabolites affected. Moreover, plant metabolic changes differed according to Azospirillum strain×cultivar combinations; indeed, 4B induced major secondary metabolic profile modifications only on Cigalon roots, while B510, probably due to its endophytic feature, induced metabolic variations on shoots and roots of both cultivars, triggering a systemic response. Plant secondary metabolite profiling thereby evidences the specific interaction between an Azospirillum strain and its original host cultivar. Copyright © 2012 Elsevier Ltd. All rights reserved.

  4. Dermal fibroblasts respond to mechanical conditioning in a strain profile dependent manner.

    PubMed

    Berry, C C; Cacou, C; Lee, D A; Bader, D L; Shelton, J C

    2003-01-01

    Fibroblasts within tissues are exposed to a dynamic mechanical environment, which influences the structural integrity of both healthy and healing soft tissues. Various systems have been proposed to subject such cells to mechanical stimulation in culture. However the diverse nature of the studies, in terms of the strain profiles and the cell types, makes direct comparisons almost impossible. The present study addresses this issue by examining the metabolic response of two cell types subjected to three well defined strain profiles.A young fibroblast cell population, represented by HuFFs, showed both greater cell proliferation and collagen production than adult dermal fibroblasts under unstrained conditions. The three strain profiles produced differing effects on both cell types. Uniaxial strains enhanced [(3)H]-thymidine incorporation for both cell types, whilst biaxial strains either inhibited or had no effect on its incorporation. In contrast, [(3)H]-proline incorporation was inhibited under biaxial and uniaxial strains for the adult fibroblasts, whilst the HuFF cells showed a small increase in proline incorporation under non-uniform and uniaxial strains.

  5. Dependence of Edge Profiles and Stability on Neutral Beam Power in NSTX

    NASA Astrophysics Data System (ADS)

    Travis, P.; Canal, G. P.; Osborne, T. H.; Maingi, R.; Sabbagh, S. A.; NSTX-U Team

    2016-10-01

    Studying the effect of neutral beam injected (NBI) power on edge plasma profiles and magnetohydrodynamic (MHD) stability is central to the understanding of edge-localized modes (ELMs). Higher heating power should quicken the development of pressure and current-driven peeling-ballooning modes. NSTX ELMy H-mode discharges with NBI power of 4, 5 and 6 MW were analyzed with a python-based set of analysis tools that fit plasma profiles, compute kinetic equilibria, and evaluate the MHD stability with the code ELITE. Electron density and temperature from Thomson scattering measurements, and ion density, temperature, and rotation from Charge Exchange Recombination Spectroscopy were inputs to the kinetic equilibrium fits. The power scan provides an opportunity to compare the stability calculations from the ELITE (ideal) and M3D-C1 (resistive) codes. Preliminary analysis shows that edge pressure profiles for the 5 and 6 MW discharges are comparable, suggesting they both reach a stability boundary. The 4 MW case shows lower edge pressure, which is likely limited by edge transport below the edge stability boundary. This work was supported in part by the U.S. Department of Energy, Office of Science, Office of Workforce Development for Teachers and Scientists (WDTS) under the Science Undergraduate Laboratory Internship (SULI) program.

  6. Network activity of mirror neurons depends on experience.

    PubMed

    Ushakov, Vadim L; Kartashov, Sergey I; Zavyalova, Victoria V; Bezverhiy, Denis D; Posichanyuk, Vladimir I; Terentev, Vasliliy N; Anokhin, Konstantin V

    2013-03-01

    In this work, the investigation of network activity of mirror neurons systems in animal brains depending on experience (existence or absence performance of the shown actions) was carried out. It carried out the research of mirror neurons network in the C57/BL6 line mice in the supervision task of swimming mice-demonstrators in Morris water maze. It showed the presence of mirror neurons systems in the motor cortex M1, M2, cingular cortex, hippocampus in mice groups, having experience of the swimming and without it. The conclusion is drawn about the possibility of the new functional network systems formation by means of mirror neurons systems and the acquisition of new knowledge through supervision by the animals in non-specific tasks.

  7. Presenilin-dependent γ-secretase activity modulates thymocyte development

    PubMed Central

    Doerfler, Petra; Shearman, Mark S.; Perlmutter, Roger M.

    2001-01-01

    In neuronal cells, presenilin-dependent γ-secretase activity cleaves amyloid precursor proteins to release Aβ peptides, and also catalyzes the release of the intracellular domain of the transmembrane receptor Notch. Accumulation of aberrant Aβ peptides appears to be causally related to Alzheimer's disease. Inhibition of Aβ peptide production is therefore a potential target for therapeutic intervention. Notch proteins play an important role in cell fate determination in many different organisms and at different stages of development, for example in mammalian T cell development. We therefore addressed whether structurally diverse γ-secretase inhibitors impair Notch function by studying thymocyte development in murine fetal thymic organ cultures. Here we show that high concentrations of the most potent inhibitors blocked thymocyte development at the most immature stage. In contrast, lower concentrations or less potent inhibitors impaired differentiation at a later stage, most notably suppressing the development of CD8 single-positive T cells. These phenotypes are consistent with an impairment of Notch signaling by γ-secretase inhibitors and define a strict Notch dose dependence of consecutive stages during thymocyte development. PMID:11470902

  8. A Solar Cycle Dependence of Nonlinearity in Magnetospheric Activity

    SciTech Connect

    Johnson, Jay R; Wing, Simon

    2005-03-08

    The nonlinear dependencies inherent to the historical K(sub)p data stream (1932-2003) are examined using mutual information and cumulant based cost as discriminating statistics. The discriminating statistics are compared with surrogate data streams that are constructed using the corrected amplitude adjustment Fourier transform (CAAFT) method and capture the linear properties of the original K(sub)p data. Differences are regularly seen in the discriminating statistics a few years prior to solar minima, while no differences are apparent at the time of solar maximum. These results suggest that the dynamics of the magnetosphere tend to be more linear at solar maximum than at solar minimum. The strong nonlinear dependencies tend to peak on a timescale around 40-50 hours and are statistically significant up to one week. Because the solar wind driver variables, VB(sub)s and dynamical pressure exhibit a much shorter decorrelation time for nonlinearities, the results seem to indicate that the nonlinearity is related to internal magnetospheric dynamics. Moreover, the timescales for the nonlinearity seem to be on the same order as that for storm/ring current relaxation. We suggest that the strong solar wind driving that occurs around solar maximum dominates the magnetospheric dynamics suppressing the internal magnetospheric nonlinearity. On the other hand, in the descending phase of the solar cycle just prior to solar minimum, when magnetospheric activity is weaker, the dynamics exhibit a significant nonlinear internal magnetospheric response that may be related to increased solar wind speed.

  9. In vivo visualization of delta opioid receptors upon physiological activation uncovers a distinct internalization profile.

    PubMed

    Faget, Lauren; Erbs, Eric; Le Merrer, Julie; Scherrer, Gregory; Matifas, Audrey; Benturquia, Nadia; Noble, Florence; Decossas, Marion; Koch, Marc; Kessler, Pascal; Vonesch, Jean-Luc; Schwab, Yannick; Kieffer, Brigitte L; Massotte, Dominique

    2012-05-23

    G-protein-coupled receptors (GPCRs) mediate numerous physiological functions and represent prime therapeutic targets. Receptor trafficking upon agonist stimulation is critical for GPCR function, but examining this process in vivo remains a true challenge. Using knock-in mice expressing functional fluorescent delta opioid receptors under the control of the endogenous promoter, we visualized in vivo internalization of this native GPCR upon physiological stimulation. We developed a paradigm in which animals were made dependent on morphine in a drug-paired context. When re-exposed to this context in a drug-free state, mice showed context-dependent withdrawal signs and activation of the hippocampus. Receptor internalization was transiently detected in a subset of CA1 neurons, uncovering regionally restricted opioid peptide release. Importantly, a pool of surface receptors always remained, which contrasts with the in vivo profile previously established for exogenous drug-induced internalization. Therefore, a distinct response is observed at the receptor level upon a physiological or pharmacological stimulation. Altogether, direct in vivo GPCR visualization enables mapping receptor stimulation promoted by a behavioral challenge and represents a powerful approach to study endogenous GPCR physiology.

  10. In vivo visualization of delta opioid receptors upon physiological activation uncovers a distinct internalization profile

    PubMed Central

    FAGET, Lauren; ERBS, Eric; LE MERRER, Julie; SCHERRER, Gregory; MATIFAS, Audrey; BENTURQUIA, Nadia; NOBLE, Florence; DECOSSAS, Marion; KOCH, Marc; KESSLER, Pascal; VONESCH, Jean-Luc; SCHWAB, Yannick; KIEFFER, Brigitte L.; MASSOTTE, Dominique

    2012-01-01

    G protein-coupled receptors (GPCRs) mediate numerous physiological functions and represent prime therapeutic targets. Receptor trafficking upon agonist stimulation is critical for GPCR function, but examining this process in vivo remains a true challenge. Using knock-in mice expressing functional fluorescent delta opioid receptors under the control of the endogenous promoter, we visualized in vivo internalization of this native GPCR upon physiological stimulation. We developed a paradigm in which animals were made dependent to morphine in a drug-paired context. When re-exposed to this context in a drug-free state, mice showed context-dependent withdrawal signs and activation of the hippocampus. Receptor internalization was transiently detected in a subset of CA1 neurons, uncovering regionally restricted opioid peptide release. Importantly, a pool of surface receptors always remained, which contrasts with the in vivo profile previously established for exogenous drug-induced internalization. Therefore, a distinct response is observed at the receptor level upon a physiological or pharmacological stimulation. Altogether, direct in vivo GPCR visualization enables mapping receptor stimulation promoted by a behavioral challenge, and represents a powerful approach to study endogenous GPCR physiology. PMID:22623675

  11. Development and Validation of a Method for Profiling Post-Translational Modification Activities Using Protein Microarrays

    PubMed Central

    Widschwendter, Martin; Sun, Dahui; Sieburg, Hans B.; Spruck, Charles

    2010-01-01

    Background Post-translational modifications (PTMs) impact on the stability, cellular location, and function of a protein thereby achieving a greater functional diversity of the proteome. To fully appreciate how PTMs modulate signaling networks, proteome-wide studies are necessary. However, the evaluation of PTMs on a proteome-wide scale has proven to be technically difficult. To facilitate these analyses we have developed a protein microarray-based assay that is capable of profiling PTM activities in complex biological mixtures such as whole-cell extracts and pathological specimens. Methodology/Principal Findings In our assay, protein microarrays serve as a substrate platform for in vitro enzymatic reactions in which a recombinant ligase, or extracts prepared from whole cells or a pathological specimen is overlaid. The reactions include labeled modifiers (e.g., ubiquitin, SUMO1, or NEDD8), ATP regenerating system, and other required components (depending on the assay) that support the conjugation of the modifier. In this report, we apply this methodology to profile three molecularly complex PTMs (ubiquitylation, SUMOylation, and NEDDylation) using purified ligase enzymes and extracts prepared from cultured cell lines and pathological specimens. We further validate this approach by confirming the in vivo modification of several novel PTM substrates identified by our assay. Conclusions/Significance This methodology offers several advantages over currently used PTM detection methods including ease of use, rapidity, scale, and sample source diversity. Furthermore, by allowing for the intrinsic enzymatic activities of cell populations or pathological states to be directly compared, this methodology could have widespread applications for the study of PTMs in human diseases and has the potential to be directly applied to most, if not all, basic PTM research. PMID:20596523

  12. Activity-Dependent Changes in Gene Expression in Schizophrenia Human-Induced Pluripotent Stem Cell Neurons.

    PubMed

    Roussos, Panos; Guennewig, Boris; Kaczorowski, Dominik C; Barry, Guy; Brennand, Kristen J

    2016-11-01

    Schizophrenia candidate genes participate in common molecular pathways that are regulated by activity-dependent changes in neurons. One important next step is to further our understanding on the role of activity-dependent changes of gene expression in the etiopathogenesis of schizophrenia. To examine whether neuronal activity-dependent changes of gene expression are dysregulated in schizophrenia. Neurons differentiated from human-induced pluripotent stem cells derived from 4 individuals with schizophrenia and 4 unaffected control individuals were depolarized using potassium chloride. RNA was extracted followed by genome-wide profiling of the transcriptome. Neurons were planted on June 21, 2013, and harvested on August 2, 2013. We performed differential expression analysis and gene coexpression analysis to identify activity-dependent or disease-specific changes of the transcriptome. Gene expression differences were assessed with linear models. Furthermore, we used gene set analyses to identify coexpressed modules that are enriched for schizophrenia risk genes. We identified 1669 genes that were significantly different in schizophrenia-associated vs control human-induced pluripotent stem cell-derived neurons and 1199 genes that are altered in these cells in response to depolarization (linear models at false discovery rate ≤0.05). The effect of activity-dependent changes of gene expression in schizophrenia-associated neurons (59 significant genes at false discovery rate ≤0.05) was attenuated compared with control samples (594 significant genes at false discovery rate ≤0.05). Using gene coexpression analysis, we identified 2 modules (turquoise and brown) that were associated with diagnosis status and 2 modules (yellow and green) that were associated with depolarization at a false discovery rate of ≤0.05. For 3 of the 4 modules, we found enrichment with schizophrenia-associated variants: brown (χ2 = 20.68; P = .002), turquoise (χ2 = 12.95; P

  13. Nanofiber diameter-dependent MAPK activity in osteoblasts.

    PubMed

    Jaiswal, Devina; Brown, Justin L

    2012-11-01

    The major challenge for bone tissue engineering lies in the fabrication of scaffolds that can mimic the extracellular matrix and promote osteogenesis. Electrospun fibers are being widely researched for this application due to high porosity, interconnectivity, and mechanical strength of the fibrous scaffolds. Electrospun poly methyl methacrylate (PMMA, 2.416 ± 0.100 μm) fibers were fabricated and etched using a 60% propylene glycol methyl ether acetate (PGMEA)/limonene (vol/vol) solution to obtain fiber diameters ranging from 2.5 to 0.5 μm in a time-dependent manner. The morphology of the fibrous scaffolds was evaluated using scanning electron microscopy and cellular compatibility with etchant-treated scaffold was assessed using immunoflurescence. Mitogen-activated protein kinases (MAPK) activation in response to different fiber diameter was evaluated with western blot as well as quantitative in-cell western. We report that electrospun micro-fibers can be etched to 0.552 ± 0.047 μm diameter without producing beads. Osteoblasts adhered to the fibers and a change in fiber diameter played a major role in modulating the activation of extracellular signal-regulated kinase (ERK) and p38 kinases with 0.882 ± 0.091 μm diameter fibers producing an inverse effect on ERK and p38 phosphorylation. These results indicate that nanofibers produced by wet etching can be effectively utilized to produce diameters that can differentially modulate MAPK activation patterns. Copyright © 2012 Wiley Periodicals, Inc.

  14. Profiles of quality of life in opiate-dependent individuals after starting methadone treatment: a latent class analysis.

    PubMed

    De Maeyer, Jessica; van Nieuwenhuizen, Chijs; Bongers, Ilja L; Broekaert, Eric; Vanderplasschen, Wouter

    2013-07-01

    This study aimed to identify classes of quality of life (QoL) among opiate-dependent individuals five to ten years after starting methadone treatment in order to tailor services to the needs of this population. A cross-sectional study of 159 opiate-dependent individuals who started outpatient methadone treatment in the region of Ghent, Belgium, between 1997 and 2002. A face-to-face structured interview was administered based on the Lancashire Quality of Life Profile, the EuropASI, Brief Symptom Inventory and the Verona Service Satisfaction Scale for Methadone Treatment. Latent class analysis was used to determine patterns of QoL. Analyses of variance and chi-square tests were used to test whether class membership was related to socio-demographic, health- and drug-related variables. Based on fit criteria, a three-class model was selected. Class Low (14.5%), 'opiate-dependent individuals living in marginal conditions', is characterised by low QoL scores on all domains. Class Intermediate (25.8%), 'stabilized, but socially excluded opiate-dependent individuals' shows high scores on the domains 'safety' and 'living situation', but low scores on all other QoL domains. Class High (59.7%), 'socially included opiate-dependent individuals', is characterised by high QoL scores on all domains, except 'finances'. The findings of this study illustrate the existence of different profiles of QoL among opiate-dependent individuals after starting methadone maintenance treatment and demonstrate the need for a continuing care approach. Insight into distinct classes of QoL can be used to design person-centred support, relevant to an individual's personal life. Copyright © 2012 Elsevier B.V. All rights reserved.

  15. Neuroserpin Differentiates Between Forms of Tissue Type Plasminogen Activator via pH Dependent Deacylation

    PubMed Central

    Carlson, Karen-Sue B.; Nguyen, Lan; Schwartz, Kat; Lawrence, Daniel A.; Schwartz, Bradford S.

    2016-01-01

    Tissue-type plasminogen activator (t-PA), initially characterized for its critical role in fibrinolysis, also has key functions in both physiologic and pathologic processes in the CNS. Neuroserpin (NSP) is a t-PA specific serine protease inhibitor (serpin) found almost exclusively in the CNS that regulates t-PA’s proteolytic activity and protects against t-PA mediated seizure propagation and blood–brain barrier disruption. This report demonstrates that NSP inhibition of t-PA varies profoundly as a function of pH within the biologically relevant pH range for the CNS, and reflects the stability, rather than the formation of NSP: t-PA acyl-enzyme complexes. Moreover, NSP differentiates between the zymogen-like single chain form (single chain t-PA, sct-PA) and the mature protease form (two chain t-PA, tct-PA) of t-PA, demonstrating different pH profiles for protease inhibition, different pH ranges over which catalytic deacylation occurs, and different pH dependent profiles of deacylation rates for each form of t-PA. NSP’s pH dependent inhibition of t-PA is not accounted for by differential acylation, and is specific for the NSP-t-PA serpin-protease pair. These results demonstrate a novel mechanism for the differential regulation of the two forms of t-PA in the CNS, and suggest a potential specific regulatory role for CNS pH in controlling t-PA proteolytic activity. PMID:27378851

  16. Routine activities and time use: a latent profile approach to sexual offenders' lifestyles.

    PubMed

    Pedneault, Amelie; Beauregard, Eric

    2014-02-01

    The routine activities of a sample of 147 incarcerated sexual offenders were studied. During interviews conducted between 1994 and 2000 in a province of Canada, data were collected on the participants' time involvement in a variety of activities in the weeks preceding their index sexual offense(s). Using latent profile analysis, a typology of lifestyles of sexual offenders was created. Five distinct profiles were identified. The inactive loner was characterized by little time involvement with their romantic partners and social activities. The social traveler spent the highest time driving and reported the highest number of friends. The single walker reported the highest amount of time walking around. The partyers' lifestyle was centered around alcohol and drugs. Finally, offenders from the familial homebody profile spent most of their time at home. Differences between profiles were investigated with regard to characteristics of index sexual offense.

  17. Regulated O2 activation in flavin-dependent monooxygenases.

    PubMed

    Frederick, Rosanne E; Mayfield, Jeffery A; DuBois, Jennifer L

    2011-08-17

    Flavin-dependent monooxygenases (FMOs) are involved in important biosynthetic pathways in diverse organisms, including production of the siderophores used for the import and storage of essential iron in serious pathogens. We have shown that the FMO from Aspergillus fumigatus, an ornithine monooxygenase (Af-OMO), is mechanistically similar to its well-studied distant homologues from mammalian liver. The latter are highly promiscuous in their choice of substrates, while Af-OMO is unusually specific. This presents a puzzle: how do Af-OMO and other FMOs of the biosynthetic classes achieve such specificity? We have discovered substantial enhancement in the rate of O(2) activation in Af-OMO in the presence of L-arginine, which acts as a small molecule regulator. Such protein-level regulation could help explain how this and related biosynthetic FMOs manage to couple O(2) activation and substrate hydroxylation to each other and to the appropriate cellular conditions. Given the essentiality of Fe to Af and the avirulence of the Af-OMO gene knock out, inhibitors of Af-OMO are likely to be drug targets against this medically intractable pathogen.

  18. Regulated O2 Activation in Flavin-Dependent Monooxygenases

    PubMed Central

    Frederick, Rosanne E.; Mayfield, Jeffery A.; DuBois, Jennifer L.

    2012-01-01

    Flavin-dependent monooxygenases (FMOs) are involved in important biosynthetic pathways in diverse organisms, including production of the siderophores used for the import and storage of essential iron in serious pathogens. We have shown that the FMO from Aspergillus fumigatus, an ornithine monooxygenase (Af-OMO), is mechanistically similar to its well-studied distant homologues from mammalian liver. The latter are highly promiscuous in their choice of substrates, while Af-OMO is unusually specific. This presents a puzzle: how do Af-OMO and other FMOs of the biosynthetic classes achieve such specificity? We have discovered substantial enhancement in the rate of O2 activation in Af-OMO in the presence of l-arginine, which acts as a small molecule regulator. Such protein-level regulation could help explain how this and related biosynthetic FMOs manage to couple O2 activation and substrate hydroxylation to each other and to the appropriate cellular conditions. Given the essentiality of Fe to Af and the avirulence of the Af-OMO gene knock out, inhibitors of Af-OMO are likely to be drug targets against this medically intractable pathogen. PMID:21774554

  19. Motor-Skill Learning Is Dependent on Astrocytic Activity.

    PubMed

    Padmashri, Ragunathan; Suresh, Anand; Boska, Michael D; Dunaevsky, Anna

    2015-01-01

    Motor-skill learning induces changes in synaptic structure and function in the primary motor cortex through the involvement of a long-term potentiation- (LTP-) like mechanism. Although there is evidence that calcium-dependent release of gliotransmitters by astrocytes plays an important role in synaptic transmission and plasticity, the role of astrocytes in motor-skill learning is not known. To test the hypothesis that astrocytic activity is necessary for motor-skill learning, we perturbed astrocytic function using pharmacological and genetic approaches. We find that perturbation of astrocytes either by selectively attenuating IP3R2 mediated astrocyte Ca(2+) signaling or using an astrocyte specific metabolic inhibitor fluorocitrate (FC) results in impaired motor-skill learning of a forelimb reaching-task in mice. Moreover, the learning impairment caused by blocking astrocytic activity using FC was rescued by administration of the gliotransmitter D-serine. The learning impairments are likely caused by impaired LTP as FC blocked LTP in slices and prevented motor-skill training-induced increases in synaptic AMPA-type glutamate receptor in vivo. These results support the conclusion that normal astrocytic Ca(2+) signaling during a reaching task is necessary for motor-skill learning.

  20. Use of focused ultrasonication in activity-based profiling of deubiquitinating enzymes in tissue.

    PubMed

    Nanduri, Bindu; Shack, Leslie A; Rai, Aswathy N; Epperson, William B; Baumgartner, Wes; Schmidt, Ty B; Edelmann, Mariola J

    2016-12-15

    To develop a reproducible tissue lysis method that retains enzyme function for activity-based protein profiling, we compared four different methods to obtain protein extracts from bovine lung tissue: focused ultrasonication, standard sonication, mortar & pestle method, and homogenization combined with standard sonication. Focused ultrasonication and mortar & pestle methods were sufficiently effective for activity-based profiling of deubiquitinases in tissue, and focused ultrasonication also had the fastest processing time. We used focused-ultrasonicator for subsequent activity-based proteomic analysis of deubiquitinases to test the compatibility of this method in sample preparation for activity-based chemical proteomics.

  1. Determination of the neutron activation profile of core drill samples by gamma-ray spectrometry.

    PubMed

    Gurau, D; Boden, S; Sima, O; Stanga, D

    2017-08-04

    This paper provides guidance for determining the neutron activation profile of core drill samples taken from the biological shield of nuclear reactors using gamma spectrometry measurements. Thus, it provides guidance for selecting a model of the right form to fit data and using least squares methods for model fitting. The activity profiles of two core samples taken from the biological shield of a nuclear reactor were determined. The effective activation depth and the total activity of core samples along with their uncertainties were computed by Monte Carlo simulation. Copyright © 2017 Elsevier Ltd. All rights reserved.

  2. Evidence for substrate-dependent inhibition profiles for human liver aldehyde oxidase.

    PubMed

    Barr, John T; Jones, Jeffrey P

    2013-01-01

    The goal of this study was to provide a reasonable assessment of how probe substrate selection may impact the results of in vitro aldehyde oxidase (AO) inhibition experiments. Here, we used a previously studied set of seven known AO inhibitors to probe the inhibition profile of a pharmacologically relevant substrate N-[(2-dimethylamino)ethyl]acridine-4-carboxamide (DACA). DACA oxidation in human liver cytosol was characterized with a measured V(max) of 2.3 ± 0.08 nmol product · min(-1) · mg(-1) and a K(m) of 6.3 ± 0.8 µM. The K(ii) and K(is) values describing the inhibition of DACA oxidation by the panel of seven inhibitors were tabulated and compared with previous findings with phthalazine as the substrate. In every case, the inhibition profile shifted to a much less uncompetitive mode of inhibition for DACA relative to phthalazine. With the exception of one inhibitor, raloxifene, this change in inhibition profile seems to be a result of a decrease in the uncompetitive mode of inhibition (an affected K(ii) value), whereas the competitive mode (K(is)) seems to be relatively consistent between substrates. Raloxifene was found to inhibit competitively when using DACA as a probe, and a previous report showed that raloxifene inhibited uncompetitively with other substrates. The relevance of these data to the mechanistic understanding of aldehyde oxidase inhibition and potential implications on drug-drug interactions is discussed. Overall, it appears that the choice in substrate may be critical when conducting mechanistic inhibition or in vitro drug-drug interactions prediction studies with AO.

  3. Evidence for Substrate-Dependent Inhibition Profiles for Human Liver Aldehyde Oxidase

    PubMed Central

    Barr, John T.

    2013-01-01

    The goal of this study was to provide a reasonable assessment of how probe substrate selection may impact the results of in vitro aldehyde oxidase (AO) inhibition experiments. Here, we used a previously studied set of seven known AO inhibitors to probe the inhibition profile of a pharmacologically relevant substrate N-[(2-dimethylamino)ethyl]acridine-4-carboxamide (DACA). DACA oxidation in human liver cytosol was characterized with a measured Vmax of 2.3 ± 0.08 nmol product · min−1 · mg−1 and a Km of 6.3 ± 0.8 µM. The Kii and Kis values describing the inhibition of DACA oxidation by the panel of seven inhibitors were tabulated and compared with previous findings with phthalazine as the substrate. In every case, the inhibition profile shifted to a much less uncompetitive mode of inhibition for DACA relative to phthalazine. With the exception of one inhibitor, raloxifene, this change in inhibition profile seems to be a result of a decrease in the uncompetitive mode of inhibition (an affected Kii value), whereas the competitive mode (Kis) seems to be relatively consistent between substrates. Raloxifene was found to inhibit competitively when using DACA as a probe, and a previous report showed that raloxifene inhibited uncompetitively with other substrates. The relevance of these data to the mechanistic understanding of aldehyde oxidase inhibition and potential implications on drug-drug interactions is discussed. Overall, it appears that the choice in substrate may be critical when conducting mechanistic inhibition or in vitro drug-drug interactions prediction studies with AO PMID:22996261

  4. Age-dependent HLA profiles of the Israeli population: impact on hematopoietic cell donor recruitment and availability.

    PubMed

    Israeli, Moshe; Oudshoorn, Machteld; Haasnoot, Geert W; Klein, Tirza; Zisser, Bracha; Bach, Gideon; Claas, Frans H J

    2014-10-01

    Approximately three million people have immigrated to the state of Israel since it was founded. Consequently, the immunogenetic profile of the younger generation may consist of a genetic mixture of formerly distinct population groups. We aimed to investigate whether HLA profiles in the Israeli population are age dependent and how this influences representation of various age groups in local donor registries. We determined HLA-A*, HLA-B*, and HLA-DRB1* low-resolution phenotypes of three age groups (n = 4,169 in each): (1) cord blood units collected between 2009 and 2013 (BABIES) and adult registry donors (2) aged 18-28 years (YOUNG) and (3) aged 49-60 years (OLD). We compared the results with virtual groups that simulate the offspring of the actual study groups. None of the three actual age groups were in Hardy-Weinberg equilibrium. The YOUNG presented four HLA-B alleles that were absent in the OLD and BABIES. A significantly higher percentage among the OLD and BABIES had a "matched" individual within their group in comparison to the YOUNG. In the YOUNG, the 10 most common haplotypes account for 16.7 % of the population, in comparison to 18.2 % in the OLD or 19.8 % in the BABIES group. The BABIES group was genetically remote from all other groups. Further disparities were found between the actual and the corresponding virtual groups. We conclude that discrete age groups in Israel present distinct immunogenetic profiles, where the younger generation is more heterogeneous. The population dynamics of the age-dependent HLA profile is multifactorial: gradual intersubgroup admixture, nonrandom mating, and entry of new alleles.

  5. Binding Activity Prediction of Cyclin-Dependent Inhibitors.

    PubMed

    Saha, Indrajit; Rak, Benedykt; Bhowmick, Shib Sankar; Maulik, Ujjwal; Bhattacharjee, Debotosh; Koch, Uwe; Lazniewski, Michal; Plewczynski, Dariusz

    2015-07-27

    The Cyclin-Dependent Kinases (CDKs) are the core components coordinating eukaryotic cell division cycle. Generally the crystal structure of CDKs provides information on possible molecular mechanisms of ligand binding. However, reliable and robust estimation of ligand binding activity has been a challenging task in drug design. In this regard, various machine learning techniques, such as Support Vector Machine, Naive Bayesian classifier, Decision Tree, and K-Nearest Neighbor classifier, have been used. The performance of these heterogeneous classification techniques depends on proper selection of features from the data set. This fact motivated us to propose an integrated classification technique using Genetic Algorithm (GA), Rotational Feature Selection (RFS) scheme, and Ensemble of Machine Learning methods, named as the Genetic Algorithm integrated Rotational Ensemble based classification technique, for the prediction of ligand binding activity of CDKs. This technique can automatically find the important features and the ensemble size. For this purpose, GA encodes the features and ensemble size in a chromosome as a binary string. Such encoded features are then used to create diverse sets of training points using RFS in order to train the machine learning method multiple times. The RFS scheme works on Principal Component Analysis (PCA) to preserve the variability information of the rotational nonoverlapping subsets of original data. Thereafter, the testing points are fed to the different instances of trained machine learning method in order to produce the ensemble result. Here accuracy is computed as a final result after 10-fold cross validation, which also used as an objective function for GA to maximize. The effectiveness of the proposed classification technique has been demonstrated quantitatively and visually in comparison with different machine learning methods for 16 ligand binding CDK docking and rescoring data sets. In addition, the best possible features

  6. Glycosylation-dependent activation of epithelial sodium channel by solnatide.

    PubMed

    Shabbir, Waheed; Tzotzos, Susan; Bedak, Minela; Aufy, Mohammad; Willam, Anita; Kraihammer, Martin; Holzner, Alexander; Czikora, Istvan; Scherbaum-Hazemi, Parastoo; Fischer, Hendrik; Pietschmann, Helmut; Fischer, Bernhard; Lucas, Rudolf; Lemmens-Gruber, Rosa

    2015-12-15

    Dysfunction of the epithelial sodium channel (ENaC), which regulates salt and water homeostasis in epithelia, causes several human pathological conditions, including pulmonary oedema. This is a potentially lethal complication of acute lung injury at least partially caused by dysfunctional alveolar liquid clearance, which in turn impairs alveolar gas exchange. Solnatide (named TIP-peptide, AP301), a 17 residue peptide mimicking the lectin-like domain of TNF has been shown to activate ENaC in several experimental animal models of acute lung injury and is being evaluated as a potential therapy for pulmonary oedema. The peptide has recently completed phase 1 and 2a clinical trials. In this study, we identify a glycosylation-dependent mechanism that preserves ENaC function and expression. Since our previous data suggested that the pore-forming subunits of ENaC are essential for maximal current activation by solnatide, we performed single- and multi-N-glycosylation site mutations in αN232,293,312,397,511Q- and δN166,211,384Q-subunits, in order to identify crucial residues for interaction with solnatide within the extracellular loop of the channel. Additionally, we generated αL576X and αN232,293,312,397,511Q,L576X deletion mutants of ENaC-α, since we have previously demonstrated that the carboxy terminal domain of this subunit is also involved in its interaction with solnatide. In cells expressing αN232,293,312,397,511Q,L576Xβγ-hENaC or δN166,311,384Q,D552Xβγ-hENaC activation by solnatide, as measured in whole cell patch clamp mode, was completely abolished, whereas it was attenuated in αL576Xβγ-hENaC- and δD552Xβγ-hENaC-expressing cells. Taken together, our findings delineate an N-glycan dependent interaction between the TIP-peptide and ENaC leading to normalization of both sodium and fluid absorption in oedematous alveoli to non-oedematous levels.

  7. Voltage-dependent potassium channels in activated rat microglia.

    PubMed Central

    Nörenberg, W; Gebicke-Haerter, P J; Illes, P

    1994-01-01

    1. Voltage-dependent currents of untreated (proliferating) and lipopolysaccharide (LPS)-treated rat microglial cells in culture were recorded using the whole-cell patch-clamp technique. 2. Membrane potentials showed prominent peaks at -35 mV and -70 mV. Membrane potentials of LPS-treated cells alternated between the two values. This may be due to a negative slope region of the I-V relation resulting in two zero current potentials. 3. From a holding potential of -70 mV, hyperpolarizing steps evoked an inwardly rectifying current both in proliferating and in LPS-treated cells, while depolarizing steps below -50 mV evoked an outwardly rectifying current only in LPS-treated microglia. The currents were K+ selective, as indicated by their reversal potential of approximately 0 mV in symmetric K+ concentrations (150 mM both intra- and extracellularly) and the reversal potential of the outward tail currents of approximately -90 mV at a normal extracellular K+ concentration (4.5 mM). 4. The activation of the outward current could be fitted by Hodgkin-Huxley-type n4 kinetics. The time constant of activation depended on voltage. 5. The inactivation of the inward and outward currents could be fitted by a single exponential. The time constant of the inward current inactivation was dependent on voltage, whereas the time constant of the outward current inactivation was virtually independent of voltage, except near the threshold of activation. Recovery of the outward from inactivation was slow and could be fitted by two exponentials. Responses to depolarizing steps were stable at 0.125 Hz, but greatly decreased from the first to the second pulse at 1 Hz. 6. The inactivation of the inward, but not of the outward, current disappeared in a low Na(+)-containing medium (5 mM). The inward current was selectively inhibited by extracellular Cs+ and Ba2+. The outward current was selectively inhibited by Cd2+, 4-aminopyridine and charybdotoxin. Replacement of intracellular K+ by an

  8. Prediction of Process-Induced Distortions in L-Shaped Composite Profiles Using Path-Dependent Constitutive Law

    NASA Astrophysics Data System (ADS)

    Ding, Anxin; Li, Shuxin; Wang, Jihui; Ni, Aiqing; Sun, Liangliang; Chang, Lei

    2016-10-01

    In this paper, the corner spring-in angles of AS4/8552 L-shaped composite profiles with different thicknesses are predicted using path-dependent constitutive law with the consideration of material properties variation due to phase change during curing. The prediction accuracy mainly depends on the properties in the rubbery and glassy states obtained by homogenization method rather than experimental measurements. Both analytical and finite element (FE) homogenization methods are applied to predict the overall properties of AS4/8552 composite. The effect of fiber volume fraction on the properties is investigated for both rubbery and glassy states using both methods. And the predicted results are compared with experimental measurements for the glassy state. Good agreement is achieved between the predicted results and available experimental data, showing the reliability of the homogenization method. Furthermore, the corner spring-in angles of L-shaped composite profiles are measured experimentally and the reliability of path-dependent constitutive law is validated as well as the properties prediction by FE homogenization method.

  9. Distinct phosphatase activity profiles in two strains of Trypanosoma cruzi.

    PubMed

    Morales-Neto, R; Hulshof, L; Ferreira, C V; Gadelha, F R

    2009-12-01

    Phosphorylation of parasite proteins plays a key role in the process of cell invasion by Trypanosoma cruzi, the etiologic agent of Chagas' disease. In this sense, characterization of parasite kinases and phosphatases could open new possibilities for the rational design of chemotherapeutic agents for the treatment of Chagas' disease. In this work, we analyzed phosphatase activities in T. cruzi homogenates from 2 strains belonging to different lineages and with different resistance to oxidative stress. Tulahuen 2 cells (Lineage I) showed higher phosphatase activities and specificity constants when compared to the Y strain (Lineage II). Tulahuen 2 had an optimum phosphatase activity at pH 4.0 and the Y strain at pH 7.0. In both cases, neutral–basic, but not acid, phosphatase activities were increased in the presence of Mg2+. Although calcium had an inhibitory effect at a pH of 7.0 and 8.0 in the Y strain, this inhibition was restricted to pH 8.0 in the other strain. Different substrates and acid phosphotyrosine and alkaline phosphatase inhibitors exhibited distinct effects on the phosphatase activity of both strains. Our results provide a better understanding of T. cruzi phosphatases and reinforce the notion of heterogeneity among T. cruzi populations.

  10. Brain reward system activity in major depression and comorbid nicotine dependence.

    PubMed

    Cardenas, Laura; Tremblay, Lescia K; Naranjo, Claudio A; Herrmann, Nathan; Zack, Martin; Busto, Usoa E

    2002-09-01

    Major depressive disorder (MDD) and nicotine dependence are highly comorbid. MDD patients may use nicotine to ameliorate depressive symptoms. The pathophysiology of the comorbidity of these two disorders is unknown. We hypothesized that a dysfunctional dopaminergic brain reward system (BRS) might be a neurobiological link between MDD and nicotine dependence and that smoking modulates the activity of the BRS by enhancing dopaminergic activity and relieving some depressive symptoms. Eighteen nicotine-dependent, nonmedicated subjects with Diagnostic and Statistical Manual of Mental Disorders (4th edition) diagnosis of MDD and 16 nicotine-dependent, control subjects participated in a double-blind, placebo-controlled, randomized parallel study. A single 30-mg oral dose of d-amphetamine (d-amph) was used to release dopamine and probe the activity of the BRS. The d-amph-mediated physiological and rewarding effects were assessed at baseline and post-treatment using standardized and validated questionnaires. Our results show that d-amph significantly increased blood pressure (p < 0.001). Subjective rewarding d-amph effects increased in both groups. Negative subjective effects were reported while on placebo during nonsmoking sessions. A significant correlation between depression severity (Hamilton depression scale) and d-amph rewarding effects was found in MDD smoker subjects (Addiction Research Center Inventory composite: r = 0.89, p < 0.000; profile of mood states composite: r = 0.71, p < 0.003; and visual analog scales composite: r = 0.78, p < 0.005). These data show that smoking did not modify the response to d-amph in MDD or control subjects, but decreased overall negative mood state during placebo sessions. Severity of depression was significantly correlated with increased rewarding effects of d-amph. Thus, although the BRS may be dysfunctional in MDD subjects, chronic nicotine use does not modify response to d-amph.

  11. Impact of physical activity on obesity and lipid profile of adults with intellectual disability.

    PubMed

    Gawlik, Krystyna; Zwierzchowska, Anna; Celebańska, Diana

    2017-09-11

    This study assessed overweight, obesity and lipid profiles in adults with intellectual disability and compared these metrics with their physical activity. Basic somatic parameters, lipid profile and weekly physical activity were examined in 27 adults with moderate intellectual disability. Chi-square independence tests and Pearson's linear correlation coefficients were used. The participants had excess body mass, excess body fat and abdominal obesity. Very high positive correlations were shown between body mass index and both waist circumference and %fat. The lipid profiles were more favourable in the general population. Healthy levels of physical activity were observed in 8% of women and 26% of men. A high negative correlation was found between physical activity and body mass index. The study group was characterized by excess body mass and insufficient levels of physical activity. Body mass index and waist circumference are sufficient indicators for identifying obesity in adults with intellectual disability. © 2017 John Wiley & Sons Ltd.

  12. Garlic sprouting is associated with increased antioxidant activity and concomitant changes in the metabolite profile.

    PubMed

    Zakarova, Alexandra; Seo, Ji Yeon; Kim, Hyang Yeon; Kim, Jeong Hwan; Shin, Jung-Hye; Cho, Kye Man; Lee, Choong Hwan; Kim, Jong-Sang

    2014-02-26

    Although garlic (Allium sativum) has been extensively studied for its health benefits, sprouted garlic has received little attention. We hypothesized that sprouting garlic would stimulate the production of various phytochemicals that improve health. Ethanolic extracts from garlic sprouted for different periods had variable antioxidant activities when assessed with in vitro assays, including the 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity assay and the oxygen radical absorbance capacity assay. Extracts from garlic sprouted for 5 days had the highest antioxidant activity, whereas extracts from raw garlic had relatively low antioxidant activity. Furthermore, sprouting changed the metabolite profile of garlic: the metabolite profile of garlic sprouted for 5-6 days was distinct from the metabolite profile of garlic sprouted for 0-4 days, which is consistent with the finding that garlic sprouted for 5 days had the highest antioxidant activity. Therefore, sprouting may be a useful way to improve the antioxidant potential of garlic.

  13. Enzymatic activities and protein profile of latex from Calotropis procera.

    PubMed

    Freitas, Cleverson Diniz T; Oliveira, Jefferson Soares; Miranda, Maria Raquel A; Macedo, Nívea Maria R; Sales, Maurício Pereira; Villas-Boas, Laurival A; Ramos, Márcio Viana

    2007-01-01

    The laticifer fluid of Calotropis procera is rich in proteins and there is evidence that they are involved in the pharmacological properties of the latex. However, not much is known about how the latex-containing proteins are produced or their functions. In this study, laticifer proteins of C. procera were pooled and examined by 1D and 2D electrophoresis, masses spectrometry (MALDI-TOF) and characterized in respect of proteolytic activity and oxidative enzymes. Soluble laticifer proteins were predominantly composed of basic proteins (PI>6.0) with molecular masses varying between 5 and 95 kDa. Proteins with a molecular mass of approximately 26,000 Da were more evident. Strong anti-oxidative activity of superoxide dismutase (EC 1.15.1.1) (1007.74+/-91.89 Ug(-1)DM) and, to a lesser extent ascorbate peroxidase (EC 1.11.1.1) (0.117(d)+/-0.013 microMol H(2)O(2)g(-1)min(-1)), were detected. However, catalase (EC 1.11.1.6) was absent. The strong proteolytic activities of laticifer proteins from C. procera were shown to be shared by at least four distinct cysteine proteinases (EC 3.4.22.16) that were isolated by gel filtration chromatography. Serine and metaloproteinases were not detected and aspartic proteinase activities were barely visible. Chitinases (EC 3.2.1.14) were also isolated in a chitin column and their activities quantified. The presence of these enzymatic activities in latex from C. procera may confirm their involvement in resistance to phytopathogens and insects, mainly in its leaves where the latex circulates abundantly.

  14. Temperature-dependent variation in API 50 CH fermentation profiles of Lactobacillus species.

    PubMed

    Nigatu, A; Ahrné, S; Molin, G

    2000-07-01

    API 50 CH fermentation profiles of 45 Lactobacillus, one Atopobium, and three Weissella strains incubated at 30 degrees C and 37 degrees C were evaluated. Atopobium uli and ten species of Lactobacillus showed stable patterns despite the change in temperature. The rest of the type strains showed discrepancy between the two incubation temperatures: 18 strains lost, 12 additionally fermented another sugar, and 7 others fermented a different one in lieu. The variation was maximum in L. delbrueckii subsp. delbrueckii. L. malefermentans failed to ferment any of the substrates at 37 degrees C. Majority of the food and plant-associated strains (mainly heterofermenters) retained distinctive traits at 30 degrees C, while most of the animal-associated strains (mostly homofermenters) did so at 37 degrees C. No general trend was observed; 30 degrees C appeared to promote heterofermentation, while 37 degrees C favored homofermenters. Use of API 50 CH profiles for taxonomic purpose in most lactobacilli appears reproducible if a specific temperature for a species is strictly followed.

  15. Depth profile by Total IBA in perovskite active layers for solar cells

    NASA Astrophysics Data System (ADS)

    Barreiros, M. A.; Alves, L. C.; Brites, M. J.; Corregidor, V.

    2017-08-01

    In recent years the record efficiency of perovskite solar cells (PSCs) has been updated exceeding now 20%. However, it is difficult to make PSCs consistently. Definite correlation has been established between the PSC performance and the perovskite film quality which involves mainly morphology, crystallinity and composition. The manufacturing development of these devices is dependent on the characterisation methodologies, on the availability of suitable and reliable analytical techniques to assess the materials composition and quality and on the relationship of these results with the cell performance. Ion beam analytical (IBA) techniques jointly with a micro-ion beam are powerful tools for materials characterisation and can provide a valuable input for the knowledge of perovskite films. Perovskite films based on CH3NH3PbI3 were prepared (from CH3NH3I and PbI2 precursors) in a planar architecture and in a mesoporous TiO2 scaffold. Proton and helium micro-beams at different energies were used in the analysis of PSC active layers, previously characterised by SEM-FEG (Scanning Electron Microscopy with a field emission gun) and XRD (X-ray diffraction). Self-consistent fit of all the obtained PIXE (Particle Induced X-ray Emission) and RBS (Rutherford Backscattering Spectrometry) spectra through Total IBA approach provided depth profiling of perovskite, its precursors and TiO2 and assess their distribution in the films. PbI2 presence and location on the active layer may hinder the charge transport and highly affect the cell performance. IBA techniques allowed to identify regions of non-uniform surface coverage and homogeneous areas and it was possible to establish the undesired presence of PbI2 and its quantitative depth profile in the planar architecture film. In the mesostructured perovskite film it was verified a non-homogeneous distribution with a decreasing of perovskite concentration down to the thin blocking layer. The good agreement between the best fits obtained

  16. Temperature dependence of mode conversion in warm, unmagnetized plasmas with a linear density profile

    SciTech Connect

    Yu, Dae Jung; Lee, Dong-Hun; Kim, Kihong

    2013-06-15

    We study theoretically the linear mode conversion between electromagnetic waves and Langmuir waves in warm, stratified, and unmagnetized plasmas, using a numerically precise calculation based on the invariant imbedding method. We verify that the principle of reciprocity for the forward and backward mode conversion coefficients holds precisely regardless of temperature. We also find that the temperature dependence of the mode conversion coefficient is substantially stronger than that previously reported. Depending on the wave frequency and the incident angle, the mode conversion coefficient is found to increase or decrease with the increase of temperature.

  17. Phytochemical profile and apoptotic activity of Onopordum cynarocephalum.

    PubMed

    Formisano, Carmen; Rigano, Daniela; Russo, Alessandra; Cardile, Venera; Caggia, Silvia; Apostolides Arnold, Nelly; Mari, Angela; Piacente, Sonia; Rosselli, Sergio; Senatore, Felice; Bruno, Maurizio

    2012-10-01

    A phytochemical investigation of acetone and chloroform extracts of the aerial parts of Onopordum cynarocephalum Boiss. et Blanche was carried out. It led to the isolation of two new sesquiterpenes, the elemane aldehyde (2) and the eudesmane (11), together with 15 known compounds: two lignans (1 and 15) and 13 sesquiterpenes (3-10, 12-14, 16, 17). The structures were elucidated by spectroscopic analyses, especially 1D and 2D NMR spectra. The anti-growth effect against three human melanoma cell lines, M14, A375, and A2058, of the different extracts and compounds of O. cynarocephalum was also investigated. Among them, the chloroform extract exhibited the strongest biological activity, while the most active compounds were the lignan arctigenin (1), and the sesquiterpenes, compounds 3, 5, and 6 belonging to the elemane type, and 7 belonging to the eudesmane type. Our data also demonstrate that acetone and chloroform extracts induce, in the A375 cell line, apoptotic cell death that could be related to an overall action of the compounds present, but in particular to the lignans arctigenin (1) and the sesquiterpenes compounds 3-8 and 16. In fact, these molecules were able to induce a high DNA fragmentation, correlated to a significant increase of the caspase-3 enzyme activity. Furthermore, apoptosis appears to be mediated, at least in part, via PTEN activity and the inhibition of Hsp70 expression.

  18. Profiling glycosyltransferase activities by tritium imaging of glycan microarrays.

    PubMed

    Serna, Sonia; Hokke, Cornelis H; Weissenborn, Martin; Flitsch, Sabine; Martin-Lomas, Manuel; Reichardt, Niels-Christian

    2013-05-10

    High-throughput microarray technology has been combined with ultrasensitive and high-resolution tritium autoradiography to create a new platform for the quantitative detection of glycosyltransferase activity on glycan arrays. In addition, we show full compatibility with the use of fluorescently labeled lectins to help with the stereochemical assignment of newly formed glycoside linkages.

  19. Beach Profiles and Transects. Ocean Related Curriculum Activities.

    ERIC Educational Resources Information Center

    Jones, Claire

    The ocean affects all of our lives. Therefore, awareness of and information about the interconnections between humans and oceans are prerequisites to making sound decisions for the future. Project ORCA (Ocean Related Curriculum Activities) has developed interdisciplinary curriculum materials designed to meet the needs of students and teachers…

  20. Fitness Profiles and Activity Patterns of Entering College Students.

    ERIC Educational Resources Information Center

    Pierce, Edgar F.; And Others

    1992-01-01

    Entering college students were evaluated for performance on maximal oxygen consumption, body composition, muscle endurance, muscle strength, and joint flexibility tests to determine the relationship of physical activity patterns to fitness levels. Results supported previous research indicating reduced fitness levels in young adults. (SM)

  1. Reproductive profile of physically active men after exhaustive endurance exercise.

    PubMed

    Vaamonde, D; Da Silva, M E; Poblador, M S; Lancho, J L

    2006-09-01

    The purpose of this study on non-professional (recreational) athletes was two-fold: 1) to determine if endurance exercise (EE) routinely used by professional athletes would produce reproductive changes in the general population, and 2) to assess reversion. Short-term exhaustive endurance exercise (EEE) can produce alterations in the hypothalamic-pituitary-gonadal (HPG) axis with subsequent fertility changes. Sixteen healthy adult male volunteers were divided into experimental (8) and control (8) groups for the exercise period. A cycloergometer provided EEE for a period of 2 weeks. The experimental group exercised four times a week; controls were without EEE. The hormonal and seminological profiles of all subjects were evaluated. Two weeks of EEE produced hormonal and seminological values in the experimental group that were statistically different from their own pre-treatment values (FSH: 3.33 +/- 1.7; LH: 3.73 +/- 1.36; sperm concentration/ml: 42.50 +/- 29.46; type a velocity: 25.23 +/- 10.9; type d velocity: 46.18 +/- 15.81; % of normal forms: 10.42 +/- 1.97) as well as from the pre- and post-treatment control group values. The measured parameters almost returned to pre-treatment levels in the experimental group 2 - 3 days after EEE ended. From this study we concluded that when subjected to EEE, individuals drawn from a recreational exercising life style experienced changes similar to those observed in studies done with athletes, and short-term EEE induced a reversible alteration to the HPG axis.

  2. Distribution of the Effect of Solar Proton Flux And Geomagnetic Activity on the Stratospheric Ozone Profile

    NASA Astrophysics Data System (ADS)

    Velinov, P. I. Y.; Tassev, Y.; Yanev, T.; Tomova, D.

    Two-way MANOVA was used to examine the impact of two factors: 1) the proton flux intensity and 2) the geomagnetic activity on the dependant variable "ozone mixing ratio" which characterizes the stratospheric ozone profiles. The examination was carried out with fixed levels of two other factors: a) the heights at which the "ozone mixing ratio" was recorded, i,e, 35 km, 30.2 km, 24.5 km, 18.4 km, 15.6 km and b) the energetic intervals within which the proton flux was measured, i.e. =0,6-4,2 MeV; 4,2-8,7 MeV; 8,7-14,5 MeV; 15-44 MeV; 39-82 MeV; 84-200 MeV; 110-500 MeV. The analysis was performed for all combinations of levels of the factors a) and b) for which data was available. It was aimed at revealing which of the factors 1) and 2) were dominating with different combinations of the factors a) and b) with fixed levels. For this purpose a post hoc analysis was performed as well. The main results are as follows: factors 1) and 2) exert statistically significant impact on the dependant variable at all of the heights examined, but not for all of energetic intervals; increase of the ozone mixing ratio was observed as a main effect of the proton flux intensity at heights 24.5 km, 18.4 km, 15.6 km, but the analysis of the simultaneous acting of factors 1) and 2) revealed a decrease of the dependant variable at these heights; these effects possibly indicate the existence of two different mechanisms of impact on the ozone mixing ratio; the afore- discussed effects decrease with the height and therefore their graphical image was named "Christmas tree".

  3. Running activity profile of adolescent tennis players during match play.

    PubMed

    Hoppe, Matthias W; Baumgart, Christian; Bornefeld, Jutta; Sperlich, Billy; Freiwald, Jürgen; Holmberg, Hans-Christer

    2014-08-01

    The aims of this study were (1) to assess the running activities of adolescent tennis players during match play with respect to velocity, acceleration, and deceleration; (2) to characterize changes in these activities during the course of a match; and (3) to identify potential differences between winners and losers. Twenty well-trained adolescent male athletes (13 ± 1 y) played one simulated match each (giving a total of 10 matches), during which distances covered at different velocity categories (0 to < 1, 1 to < 2, 2 to < 3, 3 to < 4, and ≥ 4 m·s(-1)) and number of running activities involving high velocity (≥ 3 m·s(-1)), acceleration (≥ 2 m·s(-2)), and deceleration (≤ -2 m·s(-2)) were monitored using a global positioning system (10 Hz). Heart rate was also assessed. The total match time, total distance covered, peak velocity, and mean heart rate were 81.2 ± 14.6 min, 3362 ± 869 m, 4.4 ± 0.8 m·s(-1), and 159 ± 12 beats·min(-1), respectively. Running activities involving high acceleration (0.6 ± 0.2 n·min(-1)) or deceleration (0.6 ± 0.2 n·min(-1)) were three times as frequent as those involving high velocity (0.2 ± 0.1 n·min(-1)). No change in the pattern of running activities (P ≥ .13, d ≤ 0.39) and no differences between winners and losers (P ≥ .22, d ≤ 0.53) were evident during match play. We conclude that training of well-trained adolescent male tennis players need not focus on further development of their running abilities, since this physical component of multifactorial tennis performance does not change during the course of a match and does not differ between the winners and losers.

  4. Measuring temperature-dependent activation energy in thermally activated processes: a 2D Arrhenius plot method.

    PubMed

    Li, Jian V; Johnston, Steven W; Yan, Yanfa; Levi, Dean H

    2010-03-01

    Thermally activated processes are characterized by two key quantities, activation energy (E(a)) and pre-exponential factor (nu(0)), which may be temperature dependent. The accurate measurement of E(a), nu(0), and their temperature dependence is critical for understanding the thermal activation mechanisms of non-Arrhenius processes. However, the classic 1D Arrhenius plot-based methods cannot unambiguously measure E(a), nu(0), and their temperature dependence due to the mathematical impossibility of resolving two unknown 1D arrays from one 1D experimental data array. Here, we propose a 2D Arrhenius plot method to solve this fundamental problem. Our approach measures E(a) at any temperature from matching the first and second moments of the data calculated with respect to temperature and rate in the 2D temperature-rate plane, and therefore is able to unambiguously solve E(a), nu(0), and their temperature dependence. The case study of deep level emission in a Cu(In,Ga)Se(2) solar cell using the 2D Arrhenius plot method reveals clear temperature dependent behavior of E(a) and nu(0), which has not been observable by its 1D predecessors.

  5. Cre-dependent DNA recombination activates a STING-dependent innate immune response

    PubMed Central

    Pépin, Geneviève; Ferrand, Jonathan; Höning, Klara; Jayasekara, W. Samantha N.; Cain, Jason E.; Behlke, Mark A.; Gough, Daniel J.; G. Williams, Bryan R.; Hornung, Veit; Gantier, Michael P.

    2016-01-01

    Gene-recombinase technologies, such as Cre/loxP-mediated DNA recombination, are important tools in the study of gene function, but have potential side effects due to damaging activity on DNA. Here we show that DNA recombination by Cre instigates a robust antiviral response in mammalian cells, independent of legitimate loxP recombination. This is due to the recruitment of the cytosolic DNA sensor STING, concurrent with Cre-dependent DNA damage and the accumulation of cytoplasmic DNA. Importantly, we establish a direct interplay between this antiviral response and cell–cell interactions, indicating that low cell densities in vitro could be useful to help mitigate these effects of Cre. Taking into account the wide range of interferon stimulated genes that may be induced by the STING pathway, these results have broad implications in fields such as immunology, cancer biology, metabolism and stem cell research. Further, this study sets a precedent in the field of gene-engineering, possibly applicable to other enzymatic-based genome editing technologies. PMID:27166376

  6. Brain activation profiles in dyslexic children during non-word reading: a magnetic source imaging study.

    PubMed

    Simos, P G; Breier, J I; Fletcher, J M; Foorman, B R; Bergman, E; Fishbeck, K; Papanicolaou, A C

    2000-08-18

    The purpose of the study was to identify spatiotemporal brain activation profiles associated with phonological decoding in dyslexic children using magnetic source imaging. For this purpose maps of regional cerebral activation were obtained from eleven children diagnosed with dyslexia and ten children without reading problems during engagement in a pseudoword rhyme-matching task. All dyslexic children showed aberrant activation maps consisting of reduced activity in temporoparietal areas in the left hemisphere (including the posterior part of the superior temporal, angular and supramarginal gyri) and increased activity in the right homotopic region. In contrast, the two groups of children did not differ in the degree of activity in basal temporal areas that typically precedes temporoparietal activation. This is the first study to demonstrate the existence of distinct activation profiles associated with phonological decoding in individual dyslexic children.

  7. Proteomic and activity profiles of ascorbate-glutathione cycle enzymes in germinating barley embryo.

    PubMed

    Bønsager, Birgit C; Shahpiri, Azar; Finnie, Christine; Svensson, Birte

    2010-10-01

    Enzymes involved in redox control are important during seed germination and seedling growth. Ascorbate-glutathione cycle enzymes in barley embryo extracts were monitored both by 2D-gel electrophoresis and activity measurements from 4 to 144 h post imbibition (PI). Strikingly different activity profiles were observed. No ascorbate peroxidase (APX) activity was present in mature seeds but activity was detected after 24 h PI and increased 14-fold up to 144 h PI. In contrast, dehydroascorbate reductase (DHAR) activity was present at 4h PI and first decreased by 9-fold until 72 h PI followed by a 5-fold increase at 144 h PI. Glutathione reductase and monodehydroascorbate reductase activities were also detected at 4 h PI, and showed modest increases of 1.8- and 2.7-fold, respectively, by 144 h PI. The combination of functional analysis with the proteomics approach enabled correlation of the activity profiles and protein abundance. While gel spots containing APX showed intensity changes consistent with the activity profile from 0 to 72 h PI, DHAR spot intensities indicated that post-translational regulation may be responsible for the observed changes in activity. Transcript profiling, 2D-western blotting and mass spectrometric characterization of multiple APX spots demonstrated the presence of APX1 and minor amounts of APX2.

  8. Chilean prosopis mesocarp flour: phenolic profiling and antioxidant activity.

    PubMed

    Schmeda-Hirschmann, Guillermo; Quispe, Cristina; Soriano, Maria Del Pilar C; Theoduloz, Cristina; Jiménez-Aspée, Felipe; Pérez, Maria Jorgelina; Cuello, Ana Soledad; Isla, Maria Inés

    2015-04-17

    In South America, the mesocarp flour of Prosopis species plays a prominent role as a food resource in arid areas. The aim of this work was the characterization of the phenolic antioxidants occurring in the pod mesocarp flour of Chilean Prosopis. Samples were collected in the Copiapo, Huasco and Elqui valleys from the north of Chile. The samples of P. chilensis flour exhibited a total phenolic content ranging between 0.82-2.57 g gallic acid equivalents/100 g fresh flour weight. The highest antioxidant activity, measured by the DPPH assay, was observed for samples from the Huasco valley. HPLC-MS/MS analysis allowed the tentative identification of eight anthocyanins and 13 phenolic compounds including flavonol glycosides, C-glycosyl flavones and ellagic acid derivatives. The antioxidant activity and the phenolic composition in the flour suggest that this ancient South American resource may have potential as a functional food.

  9. Simulation of the dependence of spatial fluence profiles on tissue optical properties

    NASA Astrophysics Data System (ADS)

    Miller, S.; Mitra, K.

    2016-03-01

    Medical laser applications are promoted as safe, effective treatments for a multiplicity of concerns, ranging from hyperthermal skin rejuvenation to subcutaneous tumor ablation. Chromophore and structural protein concentration and distribution within a patient's tissue vary from patient to patient and dictate the interaction of incident radiative energy of a specific wavelength with the target tissue. Laser parameters must be matched to tissue optical and thermal properties in order to achieve the desired therapeutic results without inducing unnecessary tissue damage, although accurate tissue optical properties are not always measured prior to and during laser therapies. A weighted variable step size Monte Carlo simulation of laser irradiation of skin tissue was used to determine the effects of variations in absorption (μa) and scattering coefficients (μs) and the degree of anisotropy (g) on the radiant energy transport per mm2 in response to steady-state photon propagation. The three parameters were varied in a factorial experimental design for the ranges of 0.25/mm <= μa <= 2.0/mm, 30.0/mm <= μs <= 140.0/mm, and 0.65 <= g <= 0.99 in order to isolate their impacts on the overall fluence distribution. Box plots of the resulting fluence profiles were created and compared to identify ranges in which optical property variance could be considered to significantly impact the spatial variance of fluence within the simulation volume. Results indicated that accurate prediction of the fluence profiles that will be achieved by any given medical laser treatment is unlikely without pre-treatment assessment of the tissue optical properties of individual patients.

  10. Evaluation of adaptogenic activity profile of herbal preparation.

    PubMed

    Azmathulla, Shaik; Hule, Amolkumar; Naik, Suresh Ramnath

    2006-07-01

    The herbal formulation, AVM is a proprietary formula that consists of extracts of herbs that have been used in Indian traditional medicine to promote physical and mental health, improve defense mechanisms of the body and enhance longevity. AVM (500 and 1000 mg/kg) was tested for its adaptogenic activity by determining antistress, anabolic and immunomodulatory effects. In antistress activity, pretreatment with AVM significantly attenuated the changes in ascorbic acid (from blood and adrenal), cortisol (from plasma and adrenal) and adrenal gland weights induced due to restrain stress (physical immobilization). Its antistress effect at 1000 mg/kg was comparable to that of diazepam (5 mg/kg) treated group. Leucopenia, and anemia induced by cyclophosphamide (CYP) was shown to reduce significantly by AVM. Treatment of AVM + CYP had increased spleen and thymus weights significantly as compared to CYP alone treated group. The anabolic activity was evaluated by weight gain of the levator ani muscle, ventral prostrate gland and seminal vesicles in rats as compared to untreated control.

  11. Selective functional activity measurement of a PEGylated protein with a modification-dependent activity assay.

    PubMed

    Weber, Alfred; Engelmaier, Andrea; Mohr, Gabriele; Haindl, Sonja; Schwarz, Hans Peter; Turecek, Peter L

    2017-01-05

    BAX 855 (ADYNOVATE) is a PEGylated recombinant factor VIII (rFVIII) that showed prolonged circulatory half-life compared to unmodified rFVIII in hemophilic patients. Here, the development and validation of a novel assay is described that selectively measures the activity of BAX 855 as cofactor for the serine protease factor IX, which actives factor X. This method type, termed modification-dependent activity assay, is based on PEG-specific capture of BAX 855 by an anti-PEG IgG preparation, followed by a chromogenic FVIII activity assay. The assay principle enabled sensitive measurement of the FVIII cofactor activity of BAX 855 down to the pM-range without interference by non-PEGylated FVIII. The selectivity of the capture step, shown by competition studies to primarily target the terminal methoxy group of PEG, also allowed assessment of the intactness of the attached PEG chains. Altogether, the modification-dependent activity not only enriches, but complements the group of methods to selectively, accurately, and precisely measure a PEGylated drug in complex biological matrices. In contrast to all other methods described so far, it allows measurement of the biological activity of the PEGylated protein. Data obtained demonstrate that this new method principle can be extended to protein modifications other than PEGylation and to a variety of functional activity assays. Copyright © 2016 Elsevier B.V. All rights reserved.

  12. Actuation profiles to form Zernike shapes with a thermal active mirror.

    PubMed

    Saathof, Rudolf; Schutten, Gerrit Jan M; Spronck, Jo W; Munnig Schmidt, Robert H

    2015-01-15

    In EUV lithography, the absorption of EUV light causes wavefront distortion that deteriorates the imaging process. An adaptive optics system has been developed ["Adaptive optics to counteract thermal aberrations," Ph.D. thesis (TU Delft, 2013)] to correct for this distortion using an active mirror (AM). This AM is thermally actuated by absorbing an irradiance profile exposed by a projector onto the AM. Due to thermal conductivity and bimorph-like deformation of the AM, the relation between actuation profile and actuated shape is not trivial. Therefore, this Letter describes how actuation profiles are obtained to generate Zernike shapes. These actuation profiles have been obtained by a finite-element-based optimization procedure. Furthermore, these actuation profiles are exposed to the AM, and the resulting deformations are measured. This Letter shows actuated Zernike shapes with purities higher than 0.9 for most actuation profiles. In addition, superimposed actuation profiles resulted in superimposed Zernike shapes, showing linearity needed to apply modal wavefront correction. Therefore, this approach can be used to obtain actuation profiles for this AM concept, which can be used for highly precise wavefront correction.

  13. Discovering Structure in the Space of Activation Profiles in fMRI

    PubMed Central

    Lashkari, Danial; Vul, Ed; Kanwisher, Nancy; Golland, Polina

    2009-01-01

    We present a method for discovering patterns of activation observed through fMRI in experiments with multiple stimuli/tasks. We introduce an explicit parameterization for the profiles of activation and represent fMRI time courses as such profiles using linear regression estimates. Working in the space of activation profiles, we design a mixture model that finds the major activation patterns along with their localization maps and derive an algorithm for fitting the model to the fMRI data. The method enables functional group analysis independent of spatial correspondence among subjects. We validate this model in the context of category selectivity in the visual cortex, demonstrating good agreement with prior findings based on hypothesis-driven methods. PMID:18979845

  14. Discovering structure in the space of activation profiles in fMRI.

    PubMed

    Lashkari, Danial; Vul, Ed; Kanwisher, Nancy; Golland, Polina

    2008-01-01

    We present a method for discovering patterns of activation observed through fMIRI in experiments with multiple stimuli/tasks. We introduce an explicit parameterization for the profiles of activation and represent fMRI time courses as such profiles using linear regression estimates. Working in the space of activation profiles, we design a mixture model that finds the major activation patterns along with their localization maps and derive an algorithm for fitting the model to the fMRI data. The method enables functional group analysis independent of spatial correspondence among subjects. We validate this model in the context of category selectivity in the visual cortex, demonstrating good agreement with prior findings based on hypothesis-driven methods.

  15. Substrate Activation in Flavin-Dependent Thymidylate Synthase

    PubMed Central

    2015-01-01

    Thymidylate is a critical DNA nucleotide that has to be synthesized in cells de novo by all organisms. Flavin-dependent thymidylate synthase (FDTS) catalyzes the final step in this de novo production of thymidylate in many human pathogens, but it is absent from humans. The FDTS reaction proceeds via a chemical route that is different from its human enzyme analogue, making FDTS a potential antimicrobial target. The chemical mechanism of FDTS is still not understood, and the two most recently proposed mechanisms involve reaction intermediates that are unusual in pyrimidine biosynthesis and biology in general. These mechanisms differ in the relative timing of the reaction of the flavin with the substrate. The consequence of this difference is significant: the intermediates are cationic in one case and neutral in the other, an important consideration in the construction of mechanism-based enzyme inhibitors. Here we test these mechanisms via chemical trapping of reaction intermediates, stopped-flow, and substrate hydrogen isotope exchange techniques. Our findings suggest that an initial activation of the pyrimidine substrate by reduced flavin is required for catalysis, and a revised mechanism is proposed on the basis of previous and new data. These findings and the newly proposed mechanism add an important piece to the puzzle of the mechanism of FDTS and suggest a new class of intermediates that, in the future, may serve as targets for mechanism-based design of FDTS-specific inhibitors. PMID:25025487

  16. Prevalence of human cell material: DNA and RNA profiling of public and private objects and after activity scenarios.

    PubMed

    van den Berge, M; Ozcanhan, G; Zijlstra, S; Lindenbergh, A; Sijen, T

    2016-03-01

    Especially when minute evidentiary traces are analysed, background cell material unrelated to the crime may contribute to detectable levels in the genetic analyses. To gain understanding on the composition of human cell material residing on surfaces contributing to background traces, we performed DNA and mRNA profiling on samplings of various items. Samples were selected by considering events contributing to cell material deposits in exemplary activities (e.g. dragging a person by the trouser ankles), and can be grouped as public objects, private samples, transfer-related samples and washing machine experiments. Results show that high DNA yields do not necessarily relate to an increased number of contributors or to the detection of other cell types than skin. Background cellular material may be found on any type of public or private item. When a major contributor can be deduced in DNA profiles from private items, this can be a different person than the owner of the item. Also when a specific activity is performed and the areas of physical contact are analysed, the "perpetrator" does not necessarily represent the major contributor in the STR profile. Washing machine experiments show that transfer and persistence during laundry is limited for DNA and cell type dependent for RNA. Skin conditions such as the presence of sebum or sweat can promote DNA transfer. Results of this study, which encompasses 549 samples, increase our understanding regarding the prevalence of human cell material in background and activity scenarios.

  17. Motivation and Self-Perception Profiles and Links with Physical Activity in Adolescent Girls

    ERIC Educational Resources Information Center

    Biddle, Stuart J. H.; Wang, C. K. John

    2003-01-01

    Research shows a decline in participation in physical activity across the teenage years. It is important, therefore, to examine factors that might influence adolescent girl's likelihood of being physically active. This study used contemporary theoretical perspectives from psychology to assess a comprehensive profile of motivational and…

  18. Motivation and Self-Perception Profiles and Links with Physical Activity in Adolescent Girls

    ERIC Educational Resources Information Center

    Biddle, Stuart J. H.; Wang, C. K. John

    2003-01-01

    Research shows a decline in participation in physical activity across the teenage years. It is important, therefore, to examine factors that might influence adolescent girl's likelihood of being physically active. This study used contemporary theoretical perspectives from psychology to assess a comprehensive profile of motivational and…

  19. Antioxidant activity, anti-proliferative activity, and amino acid profiles of ethanolic extracts of edible mushrooms.

    PubMed

    Panthong, S; Boonsathorn, N; Chuchawankul, S

    2016-10-17

    Biological activities of various mushrooms have recently been discovered, particularly, immunomodulatory and antitumor activities. Herein, three edible mushrooms, Auricularia auricula-judae (AA), Pleurotus abalonus (PA) and Pleurotus sajor-caju (PS) extracted using Soxhlet ethanol extraction were evaluated for their antioxidative, anti-proliferative effects on leukemia cells. Using the Folin-Ciocalteau method and Trolox equivalent antioxidant capacity assay, phenolics and antioxidant activity were found in all sample mushrooms. Additionally, anti-proliferative activity of mushroom extracts against U937 leukemia cells was determined using a viability assay based on mitochondrial activity. PA (0.5 mg/mL) and AA (0.25-0.5 mg/mL) significantly reduced cell viability. Interestingly, PS caused a hormetic-like biphasic dose-response. Low doses (0-0.25 mg/L) of PS promoted cell proliferation up to 140% relative to control, whereas higher doses (0.50 mg/mL) inhibited cell proliferation. Against U937 cells, AA IC50 was 0.28 ± 0.04 mg/mL, which was lower than PS or PA IC50 (0.45 ± 0.01 and 0.49 ± 0.001 mg/mL, respectively). Furthermore, lactate dehydrogenase (LDH) leakage conferred cytotoxicity. PS and PA were not toxic to U937 cells at any tested concentration; AA (0.50 mg/mL) showed high LDH levels and caused 50% cytotoxicity. Additionally, UPLC-HRMS data indicated several phytochemicals known to support functional activities as either antioxidant or anti-proliferative. Glutamic acid was uniquely found in ethanolic extracts of AA, and was considered an anti-cancer amino acid with potent anti-proliferative effects on U937 cells. Collectively, all mushroom extracts exhibited antioxidant effects, but their anti-proliferative effects were dose-dependent. Nevertheless, the AA extract, with highest potency, is a promising candidate for future applications.

  20. KIF4 motor regulates activity-dependent neuronal survival by suppressing PARP-1 enzymatic activity.

    PubMed

    Midorikawa, Ryosuke; Takei, Yosuke; Hirokawa, Nobutaka

    2006-04-21

    In brain development, apoptosis is a physiological process that controls the final numbers of neurons. Here, we report that the activity-dependent prevention of apoptosis in juvenile neurons is regulated by kinesin superfamily protein 4 (KIF4), a microtubule-based molecular motor. The C-terminal domain of KIF4 is a module that suppresses the activity of poly (ADP-ribose) polymerase-1 (PARP-1), a nuclear enzyme known to maintain cell homeostasis by repairing DNA and serving as a transcriptional regulator. When neurons are stimulated by membrane depolarization, calcium signaling mediated by CaMKII induces dissociation of KIF4 from PARP-1, resulting in upregulation of PARP-1 activity, which supports neuron survival. After dissociation from PARP-1, KIF4 enters into the cytoplasm from the nucleus and moves to the distal part of neurites in a microtubule-dependent manner. We suggested that KIF4 controls the activity-dependent survival of postmitotic neurons by regulating PARP-1 activity in brain development.

  1. Habitat-dependent ambient noise: Consistent spectral profiles in two African forest types

    NASA Astrophysics Data System (ADS)

    Slabbekoorn, Hans

    2004-12-01

    Many animal species use acoustic signals to attract mates, to defend territories, or to convey information that may contribute to their fitness in other ways. However, the natural environment is usually filled with competing sounds. Therefore, if ambient noise conditions are relatively constant, acoustic interference can drive evolutionary changes in animal signals. Furthermore, masking noise may cause acoustic divergence between populations of the same species if noise conditions differ consistently among habitats. In this study, ambient noise was sampled in a replicate set of sites in two habitat types in Cameroon: contiguous rainforest and ecotone forest patches north of the rainforest. The noise characteristics of the two forest types show significant and consistent differences. Multiple samples taken at two rainforest sites in different seasons vary little and remain distinct from those in ecotone forest. The rainforest recordings show many distinctive frequency bands, with a general increase in amplitude from low to high frequencies. Ecotone forest only shows a distinctive high-frequency band at some parts of the day. Habitat-dependent abiotic and biotic sound sources and to some extent habitat-dependent sound transmission are the likely causes of these habitat-dependent noise spectra. .

  2. Homeostatic Activity-Dependent Tuning of Recurrent Networks for Robust Propagation of Activity.

    PubMed

    Gjorgjieva, Julijana; Evers, Jan Felix; Eglen, Stephen J

    2016-03-30

    Developing neuronal networks display spontaneous bursts of action potentials that are necessary for circuit organization and tuning. While spontaneous activity has been shown to instruct map formation in sensory circuits, it is unknown whether it plays a role in the organization of motor networks that produce rhythmic output. Using computational modeling, we investigate how recurrent networks of excitatory and inhibitory neuronal populations assemble to produce robust patterns of unidirectional and precisely timed propagating activity during organism locomotion. One example is provided by the motor network inDrosophilalarvae, which generates propagating peristaltic waves of muscle contractions during crawling. We examine two activity-dependent models, which tune weak network connectivity based on spontaneous activity patterns: a Hebbian model, where coincident activity in neighboring populations strengthens connections between them; and a homeostatic model, where connections are homeostatically regulated to maintain a constant level of excitatory activity based on spontaneous input. The homeostatic model successfully tunes network connectivity to generate robust activity patterns with appropriate timing relationships between neighboring populations. These timing relationships can be modulated by the properties of spontaneous activity, suggesting its instructive role for generating functional variability in network output. In contrast, the Hebbian model fails to produce the tight timing relationships between neighboring populations required for unidirectional activity propagation, even when additional assumptions are imposed to constrain synaptic growth. These results argue that homeostatic mechanisms are more likely than Hebbian mechanisms to tune weak connectivity based on spontaneous input in a recurrent network for rhythm generation and robust activity propagation. How are neural circuits organized and tuned to maintain stable function and produce robust output

  3. Enzyme Activity Profiles during Fruit Development in Tomato Cultivars and Solanum pennellii1[W][OA

    PubMed Central

    Steinhauser, Marie-Caroline; Steinhauser, Dirk; Koehl, Karin; Carrari, Fernando; Gibon, Yves; Fernie, Alisdair R.; Stitt, Mark

    2010-01-01

    Enzymes interact to generate metabolic networks. The activities of more than 22 enzymes from central metabolism were profiled during the development of fruit of the modern tomato cultivar Solanum lycopersicum ‘M82’ and its wild relative Solanum pennellii (LA0716). In S. pennellii, the mature fruit remains green and contains lower sugar and higher organic acid levels. These genotypes are the parents of a widely used near introgression line population. Enzymes were also profiled in a second cultivar, S. lycopersicum ‘Moneymaker’, for which data sets for the developmental changes of metabolites and transcripts are available. Whereas most enzyme activities declined during fruit development in the modern S. lycopersicum cultivars, they remained high or even increased in S. pennellii, especially enzymes required for organic acid synthesis. The enzyme profiles were sufficiently characteristic to allow stages of development and cultivars and the wild species to be distinguished by principal component analysis and clustering. Many enzymes showed coordinated changes during fruit development of a given genotype. Comparison of the correlation matrices revealed a large overlap between the two modern cultivars and considerable overlap with S. pennellii, indicating that despite the very different development responses, some basic modules are retained. Comparison of enzyme activity, metabolite profiles, and transcript profiles in S. lycopersicum ‘Moneymaker’ revealed remarkably little connectivity between the developmental changes of transcripts and enzymes and even less between enzymes and metabolites. We discuss the concept that the metabolite profile is an emergent property that is generated by complex network interactions. PMID:20335402

  4. Inversion for range-dependent water column sound speed profiles on the New Jersey shelf using a linearized perturbative method.

    PubMed

    Ballard, Megan S; Becker, Kyle M

    2010-06-01

    The environment of the New Jersey shelf is characterized by high spatial and temporal variability of water column properties caused by intrusions of warm, salty water from the continental slope. These intrusions cause fluctuations in the water column sound speed profile which can have significant effects on acoustic propagation in shallow water. In this work, a linearized perturbative inverse technique is applied to estimate range-dependent water column sound speed profiles. This method utilizes estimates of horizontal wave numbers to determine sound speed as a function of depth. This technique is appropriate for the range-dependent shallow-water environment as horizontal wave numbers can be measured semilocally (1-2 km aperture) and their values are a direct measurement of the local environmental parameters. Difficulty is encountered in application of the perturbative inverse technique because the wave number data are insensitive to some portions of the waveguide and, as a result, the solution can deviate wildly from true values. This issue is addressed by application of approximate equality constraints which force the solution to be close to likely values at prescribed locations.

  5. Protein Kinase Activity of Phosphoinositide 3-Kinase Regulates Cytokine-Dependent Cell Survival

    PubMed Central

    Green, Benjamin D.; Barry, Emma F.; Ma, Yuefang; Woodcock, Joanna; Fitter, Stephen; Zannettino, Andrew C. W.; Pitson, Stuart M.; Hughes, Timothy P.; Lopez, Angel F.; Shepherd, Peter R.; Wei, Andrew H.; Ekert, Paul G.; Guthridge, Mark A.

    2013-01-01

    The dual specificity protein/lipid kinase, phosphoinositide 3-kinase (PI3K), promotes growth factor-mediated cell survival and is frequently deregulated in cancer. However, in contrast to canonical lipid-kinase functions, the role of PI3K protein kinase activity in regulating cell survival is unknown. We have employed a novel approach to purify and pharmacologically profile protein kinases from primary human acute myeloid leukemia (AML) cells that phosphorylate serine residues in the cytoplasmic portion of cytokine receptors to promote hemopoietic cell survival. We have isolated a kinase activity that is able to directly phosphorylate Ser585 in the cytoplasmic domain of the interleukin 3 (IL-3) and granulocyte macrophage colony stimulating factor (GM-CSF) receptors and shown it to be PI3K. Physiological concentrations of cytokine in the picomolar range were sufficient for activating the protein kinase activity of PI3K leading to Ser585 phosphorylation and hemopoietic cell survival but did not activate PI3K lipid kinase signaling or promote proliferation. Blockade of PI3K lipid signaling by expression of the pleckstrin homology of Akt1 had no significant impact on the ability of picomolar concentrations of cytokine to promote hemopoietic cell survival. Furthermore, inducible expression of a mutant form of PI3K that is defective in lipid kinase activity but retains protein kinase activity was able to promote Ser585 phosphorylation and hemopoietic cell survival in the absence of cytokine. Blockade of p110α by RNA interference or multiple independent PI3K inhibitors not only blocked Ser585 phosphorylation in cytokine-dependent cells and primary human AML blasts, but also resulted in a block in survival signaling and cell death. Our findings demonstrate a new role for the protein kinase activity of PI3K in phosphorylating the cytoplasmic tail of the GM-CSF and IL-3 receptors to selectively regulate cell survival highlighting the importance of targeting such pathways in

  6. The Activity Profile of Elite Low-Kick Kickboxing Competition.

    PubMed

    Slimani, Maamer; Chaabene, Helmi; Miarka, Bianca; Chamari, Karim

    2017-02-01

    To determine the performance aspects (time-motion and technical-tactical analysis) of top-level low-kick kickboxers according to gender, weight category, combat round, and match outcome. Seventy-two kickboxers (44 male, 28 female) were studied. Thirty-six bouts (male = 61, female = 41 rounds) were analyzed using a time-motion system. Time structure was classified into 3 phases: preparatory-activity time (PT), fighting time (FT), and stoppage time (ST). Referee decisions caused an overall effort:pause ratio (E:P) of ~1:1.5, with a significant difference between weight categories (light and middleweights = 1:1.5, heavyweight = 1:1). This ratio was ~1:6 when high-intensity actions-to-pause activities were considered. Significant differences were also observed between rounds (all P < .001), with 1st-, 2nd-, and 3rd-round E:Ps of 1:1, 1:1.5, and ~1:2, respectively. The relative times of FT and PT, total attacking actions, upper-limb actions, number of technical actions performed on the head, and the number of high-intensity actions were higher in males than females (all P = .05). Males performed more jab-cross actions and fewer low kicks than females (P < .001). Males used upper-limb (63.4%) more than lower-limb techniques (36.6%), targeting the head (56.9%) more than the body/leg (43.1%), with no significant difference from females (P > .05). E:P was similar between winners and losers. However, the numbers of technical actions performed on the head, counterattack actions, jab-cross technique, and total punches were higher in winners than losers (all P < .05). Training programs need to be adapted to the specific requirements of kickboxers' weight categories and gender to develop the technical-tactical abilities that improve athletes' chances of winning.

  7. Dependency of Tearing Mode Stability on Current and Pressure Profiles in DIII-D Hybrid Discharges

    NASA Astrophysics Data System (ADS)

    Kim, K.; Park, J. M.; Murakami, M.; La Haye, R. J.; Na, Y.-S.; SNU/ORAU; ORNL; Atomics, General; SNU; DIII-D Team

    2016-10-01

    Understanding the physics of the onset and evolution of tearing modes (TMs) in tokamak plasmas is important for high- β steady-state operation. Based on DIII-D steady-state hybrid experiments with accurate equilibrium reconstruction and well-measured plasma profiles, the 2/1 tearing mode can be more stable with increasing local current and pressure gradient at rational surface and with lower pressure peaking and plasma inductance. The tearing stability index Δ', estimated by the Rutherford equation with experimental mode growth rate was validated against Δ' calculated by linear eigenvalue solver (PEST3); preliminary comprehensive MHD modeling by NIMROD reproduced the TM onset reasonably well. We present a novel integrated modeling for the purpose of predicting TM onset in experiment by combining a model equilibrium reconstruction using IPS/FASTRAN, linear stability Δ' calculation using PEST3, and fitting formula for critical Δ' from NIMROD. Work supported in part by the US DoE under DE-AC05-06OR23100, DE-AC05-00OR22725, and DEFC02-04ER54698.

  8. A peroxisome proliferator-activated receptor gamma agonist influenced daily profile of energy expenditure in genetically obese diabetic rats.

    PubMed

    Yoshida, Yuki; Ichikawa, Mineko; Ohta, Minoru; Kanai, Setsuko; Kobayash, Mikako; Ichimaru, Yuhei; Shimazoe, Takao; Watanabe, Shigenori; Funakoshi, Akihiro; Miyasak, Kyoko

    2002-03-01

    Otsuka Long Evans Tokushima Fatty (OLETF) rats were developed as a model of non-insulin-dependent diabetes mellitus (NIDDM) with mild obesity. We reported that the daily profiles of energy expenditure associated with two peaks (one between 05:00 and 08:00 and the other between 20:00 and 22:00) were observed at 8 weeks of age (without NIDDM), while these two peaks disappeared at 24 weeks of age with NIDDM. As a new anti-diabetic drug, a peroxisome proliferator-activated receptor y agonist pioglitazone hydrochloride has been developed, we examined whether pioglitazone normalized daily profiles of energy expenditure at 24 weeks of age. A control diet and pioglitazone (0.1%)-containing diet were fed from 6 weeks of age. The two peaks of daily profiles of energy expenditure, which disappeared in OLETF rats with the control diet at 24 weeks of age, were reproduced by administration of pioglitazone. The respiratory quotient was lower and fat derived energy used for combustion was increased by pioglitazone at both ages. The body weight, daily food intake, plasma levels of fat, insulin, leptin and the wet weight of visceral fat were not influenced, but the levels of blood hemoglobin Alc and plasma tumor necrosis factor a were decreased by pioglitazone. Administration of pioglitazone improved daily profiles of energy expenditure via affecting glucose and fat metabolisms.

  9. Phytochemical profiles and antioxidant activity of brown rice varieties.

    PubMed

    Gong, Er Sheng; Luo, Shun Jing; Li, Tong; Liu, Cheng Mei; Zhang, Guo Wen; Chen, Jun; Zeng, Zi Cong; Liu, Rui Hai

    2017-07-15

    The phytochemical content and antioxidant activity of eight varieties of brown rice (BR) are reported. The total phenolic contents of BR ranged from 72.45 to 120.13mg of gallic acid equiv./100g. The phenolics from bound fraction contributed 40.6-50.2% of the total phenolic content. The total flavonoid contents of BR ranged from 75.90 to 112.03mg catechin equiv./100g. The flavonoids from the bound fraction contributed 26.9-48.2% of total flavonoids. Trans-ferulic acid was the predominant phenolic acid in BR. Total trans-ferulic acid content ranged from 161.42 to 374.81μg/100g. The percentage of trans-ferulic acid in bound fraction ranged from 96.4% to 99.2%. Only α- and γ-tocopherols and -tocotrienols were detected in BR with α-tocopherol and γ-tocotrienol being the predominant. The total peroxyl radical scavenging capacity (PSC) of BR ranged from 18.29 to 40.33mg vitamin C equiv./100g. The bound fraction contributed 67.2-77.2% of total PSC. Copyright © 2017 Elsevier Ltd. All rights reserved.

  10. Profiles of Executive Functioning: Associations with Substance Dependence and Risky Sexual Behavior

    PubMed Central

    Golub, Sarit A.; Starks, Tyrel J.; Kowalczyk, William J.; Thompson, Louisa I.; Parsons, Jeffrey T.

    2012-01-01

    The present investigations applied a theoretical perspective regarding the impact of executive functioning (EF) on sexual risk among substance users, by using a methodological approach designed to examine whether EF subtypes differentially predict behavior patterns. Participants included 104 substance-using HIV-negative gay and bisexual men. Participants completed five neuropsychological assessment tasks selected to tap discrete EF components, and these data were linked to data on substance dependence and behavioral reports of substance use and sexual risk in the past 30 days. Cluster analysis identified three EF subtypes: a) High-performing (good performance across all measures); b) Low Performing (poor performance across all measures); and c) Poor IGT Performance (impairment on the Iowa Gambling Task (IGT) and its variant, but good performance on all other tasks). The three subtypes did not differ in amount of substance use, but the Low-Performing subtype was associated with greater rates of substance dependence. The Low-Performing subtype reported the highest rates of sexual behavior and risk, while the Poor IGT-Performance subtype reported the lowest rates of sexual risk-taking. Global associations between substance use and sexual risk were strongest among the Low-Performing subtype, but event-level associations appeared strongest among individuals in the High-Performing subtype. These data suggest complex associations between EF and sexual risk among substance users, and suggest that the relationship between substance use and sexual risk may vary by EF subtypes. PMID:22775771

  11. Profiles of executive functioning: associations with substance dependence and risky sexual behavior.

    PubMed

    Golub, Sarit A; Starks, Tyrel J; Kowalczyk, William J; Thompson, Louisa I; Parsons, Jeffrey T

    2012-12-01

    The present investigations applied a theoretical perspective regarding the impact of executive functioning (EF) on sexual risk among substance users, using a methodological approach designed to examine whether EF subtypes differentially predict behavior patterns. Participants included 104 substance-using HIV-negative gay and bisexual men. Participants completed 5 neuropsychological assessment tasks selected to tap discrete EF components, and these data were linked to data on substance dependence and behavioral reports of substance use and sexual risk in the past 30 days. Cluster analysis identified 3 EF subtypes: (a) high performing (good performance across all measures); (b) low performing (poor performance across all measures); and (c) poor IGT performance (impairment on the Iowa Gambling Task [IGT] and its variant, but good performance on all other tasks). The 3 subtypes did not differ in amount of substance use, but the low-performing subtype was associated with greater rates of substance dependence. The low-performing subtype reported the highest rates of sexual behavior and risk, while the poor-IGT-performance subtype reported the lowest rates of sexual risk taking. Global associations between substance use and sexual risk were strongest among the low-performing subtype, but event-level associations appeared strongest among individuals in the high-performing subtype. These data suggest complex associations between EF and sexual risk among substance users, and suggest that the relationship between substance use and sexual risk may vary by EF subtypes. 2013 APA, all rights reserved

  12. Milestone report: Status report on time-dependent modeling for current profile feedback control

    SciTech Connect

    Casper, T.A.; Crotinger, J.; Haney, S.

    1995-09-29

    During the past year, LLNL efforts in the DIII-D experimental program have expanded to include time-dependent modeling of advanced tokamak (AT) operating modes. Consistent with our involvement in experimental operations, we have directed our initial efforts at modeling the negative central shear (NCS) configuration, an important and attractive mode of operation for reducing the size and cost of future tokamak experiments without sacrificing performance. In this endeavor, we have brought into use the Corsica modeling code as a tool for investigating the time-dependent evolution and control of various operating modes. In our current efforts, we are contributing to the analysis of the NCS experimental data using analysis tools such as the EFIT equilibrium code and the ONETWO and TRANSP transport codes. Results of these analyses are being used for comparisons with the Corsica modeling. Future directions include the modeling of startup and sustaining of NCS (and other AT) configurations, the understanding of current drive effects, the development of current drive scenarios and control algorithms, and the design of experiments and prediction of experimental results. We are currently in the early stages of applying this powerful modeling tool to the DIII-D experimental program.

  13. Activity-Based Protein Profiling of Ammonia Monooxygenase in Nitrosomonas europaea.

    PubMed

    Bennett, Kristen; Sadler, Natalie C; Wright, Aaron T; Yeager, Chris; Hyman, Michael R

    2016-04-01

    Nitrosomonas europaea is an aerobic nitrifying bacterium that oxidizes ammonia (NH3) to nitrite (NO2 (-)) through the sequential activities of ammonia monooxygenase (AMO) and hydroxylamine dehydrogenase (HAO). Many alkynes are mechanism-based inactivators of AMO, and here we describe an activity-based protein profiling method for this enzyme using 1,7-octadiyne (17OD) as a probe. Inactivation of NH4 (+)-dependent O2 uptake by N. europaea by 17OD was time- and concentration-dependent. The effects of 17OD were specific for ammonia-oxidizing activity, andde novoprotein synthesis was required to reestablish this activity after cells were exposed to 17OD. Cells were reacted with Alexa Fluor 647 azide using a copper-catalyzed azide-alkyne cycloaddition (CuAAC) (click) reaction, solubilized, and analyzed by SDS-PAGE and infrared (IR) scanning. A fluorescent 28-kDa polypeptide was observed for cells previously exposed to 17OD but not for cells treated with either allylthiourea or acetylene prior to exposure to 17OD or for cells not previously exposed to 17OD. The fluorescent polypeptide was membrane associated and aggregated when heated with β-mercaptoethanol and SDS. The fluorescent polypeptide was also detected in cells pretreated with other diynes, but not in cells pretreated with structural homologs containing a single ethynyl functional group. The membrane fraction from 17OD-treated cells was conjugated with biotin-azide and solubilized in SDS. Streptavidin affinity-purified polypeptides were on-bead trypsin-digested, and amino acid sequences of the peptide fragments were determined by liquid chromatography-mass spectrometry (LC-MS) analysis. Peptide fragments from AmoA were the predominant peptides detected in 17OD-treated samples. In-gel digestion and matrix-assisted laser desorption ionization-tandem time of flight (MALDI-TOF/TOF) analyses also confirmed that the fluorescent 28-kDa polypeptide was AmoA. Copyright © 2016, American Society for Microbiology. All

  14. Activity-Based Protein Profiling of Ammonia Monooxygenase in Nitrosomonas europaea

    SciTech Connect

    Bennett, Kristen; Sadler, Natalie C.; Wright, Aaron T.; Yeager, Chris; Hyman, Michael R.; Löffler, F. E.

    2016-01-29

    Nitrosomonas europaeais an aerobic nitrifying bacterium that oxidizes ammonia (NH3) to nitrite (NO2) through the sequential activities of ammonia monooxygenase (AMO) and hydroxylamine dehydrogenase (HAO). Many alkynes are mechanism-based inactivators of AMO, and here we describe an activity-based protein profiling method for this enzyme using 1,7-octadiyne (17OD) as a probe. Inactivation of NH4+-dependent O2uptake byN. europaeaby 17OD was time- and concentration-dependent. The effects of 17OD were specific for ammonia-oxidizing activity, andde novoprotein synthesis was required to reestablish this activity after cells were exposed to 17OD. Cells were reacted with Alexa Fluor 647 azide using a copper-catalyzed azide-alkyne cycloaddition (CuAAC) (click) reaction, solubilized, and analyzed by SDS-PAGE and infrared (IR) scanning. A fluorescent 28-kDa polypeptide was observed for cells previously exposed to 17OD but not for cells treated with either allylthiourea or acetylene prior to exposure to 17OD or for cells not previously exposed to 17OD. The fluorescent polypeptide was membrane associated and aggregated when heated with β-mercaptoethanol and SDS. The fluorescent polypeptide was also detected in cells pretreated with other diynes, but not in cells pretreated with structural homologs containing a single ethynyl functional group. The membrane fraction from 17OD-treated cells was conjugated with biotin-azide and solubilized in SDS. Streptavidin affinity-purified polypeptides were on-bead trypsin-digested, and amino acid sequences of the peptide fragments were determined by liquid chromatography-mass spectrometry (LC-MS) analysis. Peptide fragments from AmoA were the predominant peptides detected in 17OD-treated samples. In-gel digestion and matrix

  15. Obscuration-dependent Evolution of Active Galactic Nuclei

    NASA Astrophysics Data System (ADS)

    Buchner, Johannes; Georgakakis, Antonis; Nandra, Kirpal; Brightman, Murray; Menzel, Marie-Luise; Liu, Zhu; Hsu, Li-Ting; Salvato, Mara; Rangel, Cyprian; Aird, James; Merloni, Andrea; Ross, Nicholas

    2015-04-01

    We aim to constrain the evolution of active galactic nuclei (AGNs) as a function of obscuration using an X-ray-selected sample of ~2000 AGNs from a multi-tiered survey including the CDFS, AEGIS-XD, COSMOS, and XMM-XXL fields. The spectra of individual X-ray sources are analyzed using a Bayesian methodology with a physically realistic model to infer the posterior distribution of the hydrogen column density and intrinsic X-ray luminosity. We develop a novel non-parametric method that allows us to robustly infer the distribution of the AGN population in X-ray luminosity, redshift, and obscuring column density, relying only on minimal smoothness assumptions. Our analysis properly incorporates uncertainties from low count spectra, photometric redshift measurements, association incompleteness, and the limited sample size. We find that obscured AGNs with N H > 1022 cm-2 account for {77}+4-5% of the number density and luminosity density of the accretion supermassive black hole population with L X > 1043 erg s-1, averaged over cosmic time. Compton-thick AGNs account for approximately half the number and luminosity density of the obscured population, and {38}+8-7% of the total. We also find evidence that the evolution is obscuration dependent, with the strongest evolution around N H ≈ 1023 cm-2. We highlight this by measuring the obscured fraction in Compton-thin AGNs, which increases toward z ~ 3, where it is 25% higher than the local value. In contrast, the fraction of Compton-thick AGNs is consistent with being constant at ≈35%, independent of redshift and accretion luminosity. We discuss our findings in the context of existing models and conclude that the observed evolution is, to first order, a side effect of anti-hierarchical growth.

  16. The effect of dietary fat content on phospholipid fatty acid profile is muscle fiber type dependent.

    PubMed

    Janovská, Alena; Hatzinikolas, George; Mano, Mark; Wittert, Gary A

    2010-04-01

    A high-saturated-fat diet (HFD) induces obesity and insulin resistance (IR). IR has been linked to alterations and increased saturation in the phospholipid composition of skeletal muscles. We aimed to determine whether HFD feeding affects fatty acid (FA) membrane profile in a muscle fiber type-specific manner. We measured phospholipid FAs and expression of FA synthesis genes in oxidative soleus (SOL) and glycolytic extensor digitorum longus (EDL) muscles from rats fed either standard chow (standard laboratory diet, SLD) or a HFD. The HFD increased fat mass, plasma insulin, and leptin levels. Compared with EDL, SOL muscles preferentially accumulated C18 over C16 FAs and n-6 over n-3 polyunsaturated FAs (PUFAs) on either diet. With the HFD, SOL muscles contained more n-9 monounsaturated FAs (MUFAs) and n-6 PUFAs and less n-7 MUFAs and n-3 PUFAs than EDL muscles and had lower unsaturation index, a pattern known to be associated with IR. Stearoyl-CoA desaturase-1 expression was approximately 13-fold greater in EDL than in SOL muscles but did not change with the HFD in either muscle. The expression of Elongase-5 was higher, and that of Elongase-6 (Elovl6) was lower in EDL compared with SOL muscles with both diets. In EDL muscles, the expression of Elovl6 was lower in the HFD than in the SLD. The pattern of FA uptake, expression, and diet-induced changes in FA desaturating and elongating enzymes maintained higher FA unsaturation in EDL muscles. Accordingly, the fiber type composition of skeletal muscles and their distribution may be important in the development and progression of obesity and IR.

  17. Senescence associated secretory phenotype profile from primary lung mice fibroblasts depends on the senescence induction stimuli.

    PubMed

    Maciel-Barón, L A; Morales-Rosales, S L; Aquino-Cruz, A A; Triana-Martínez, F; Galván-Arzate, S; Luna-López, A; González-Puertos, V Y; López-Díazguerrero, N E; Torres, C; Königsberg, Mina

    2016-02-01

    Cellular senescence is a multifactorial phenomenon of growth arrest and distorted function, which has been recognized as an important feature during tumor suppression mechanisms and a contributor to aging. Senescent cells have an altered secretion pattern called Senescence-Associated Secretory Phenotype (SASP) that comprises a complex mix of factors including cytokines, growth factors, chemokines, and matrix metalloproteinases. SASP has been related with local inflammation that leads to cellular transformation and neurodegenerative diseases. Various pathways for senescence induction have been proposed; the most studied is replicative senescence due to telomere attrition called replicative senescence (RS). However, senescence can be prematurely achieved when cells are exposed to diverse stimuli such as oxidative stress (stress-induced premature senescence, SIPS) or proteasome inhibition (proteasome inhibition-induced premature senescence, PIIPS). SASP has been characterized in RS and SIPS but not in PIIPS. Hence, our aim was to determine SASP components in primary lung fibroblasts obtained from CD-1 mice induced to senescence by PIIPS and compare them to RS and SIPS. Our results showed important variations in the 62 cytokines analyzed, while SIPS and RS showed an increase in the secretion of most cytokines, and in PIIPS only 13 were incremented. Variations in glutathione-redox balance were also observed in SIPS and RS, and not in PIIPS. All senescence types SASP displayed a pro-inflammatory profile and increased proliferation in L929 mice fibroblasts exposed to SASP. However, the behavior observed was not exactly the same, suggesting that the senescence induction pathway might encompass dissimilar responses in adjacent cells and promote different outcomes.

  18. Distribution of Metabolically Active Prokaryotes (Archaea and Bacteria) throughout the Profiles of Chernozem and Brown Semidesert Soil

    NASA Astrophysics Data System (ADS)

    Semenov, M. V.; Manucharova, N. A.; Stepanov, A. L.

    2016-02-01

    The distribution of metabolically active cells of archaea and bacteria in the profiles of typical chernozems (Voronezh oblast) and brown semidesert soils (Astrakhan oblast) of natural and agricultural ecosystems was studied using the method of fluorescent in situ hybridization (FISH). The studied soils differed sharply in the microbial biomass and in the numbers of metabolically active cells of archaea and bacteria. The number of active bacterial cells was 3.5-7.0 times greater than that of archaea. In the arable chernozem, the numbers of active cells of archaea and bacteria were 2.6 and 1.5 times, respectively, lower than those in the chernozem under the shelterbelt. The agricultural use of the brown semidesert soil had little effect on the abundances of bacteria and archaea. The soil organic carbon content was the major factor controlling the numbers of metabolically active cells of both domains. However, the dependence of the abundance of bacteria on the organic matter content was more pronounced. The decrease in the organic carbon and total nitrogen contents down the soil profiles was accompanied by the decrease in the bacteria: archaea ratio attesting to a better adaptation of archaea to the permanent deficiency of carbon and nitrogen. The bacteria: archaea ratio can serve as an ecotrophic indicator of the state of soil microbial communities.

  19. Phytochemical profile of Rosmarinus officinalis and Salvia officinalis extracts and correlation to their antioxidant and anti-proliferative activity.

    PubMed

    Kontogianni, Vassiliki G; Tomic, Goran; Nikolic, Ivana; Nerantzaki, Alexandra A; Sayyad, Nisar; Stosic-Grujicic, Stanislava; Stojanovic, Ivana; Gerothanassis, Ioannis P; Tzakos, Andreas G

    2013-01-01

    The goal of this study was to monitor the anti-proliferative activity of Rosmarinus officinalis and Salvia officinalis extracts against cancer cells and to correlate this activity with their phytochemical profiles using liquid chromatography/diode array detection/electrospray ion trap tandem mass spectrometry (LC/DAD/ESI-MS(n)). For the quantitative estimation of triterpenic acids in the crude extracts an NMR based methodology was used and compared with the HPLC measurements, both applied for the first time, for the case of betulinic acid. Both extracts exerted cytotoxic activity through dose-dependent impairment of viability and mitochondrial activity of rat insulinoma m5F (RINm5F) cells. Decrease of RINm5F viability was mediated by nitric oxide (NO)-induced apoptosis. Importantly, these extracts potentiated NO and TNF-α release from macrophages therefore enhancing their cytocidal action. The rosemary extract developed more pronounced antioxidant, cytotoxic and immunomodifying activities, probably due to the presence of betulinic acid and a higher concentration of carnosic acid in its phytochemical profile.

  20. Dose- and time-dependent effects of phenobarbital on gene expression profiling in human hepatoma HepaRG cells

    SciTech Connect

    Lambert, Carine B.; Spire, Catherine Claude, Nancy; Guillouzo, Andre

    2009-02-01

    Phenobarbital (PB) induces or represses a wide spectrum of genes in rodent liver. Much less is known about its effects in human liver. We used pangenomic cDNA microarrays to analyze concentration- and time-dependent gene expression profile changes induced by PB in the well-differentiated human HepaRG cell line. Changes in gene expression profiles clustered at specific concentration ranges and treatment times. The number of correctly annotated genes significantly modulated by at least three different PB concentration ranges (spanning 0.5 to 3.2 mM) at 20 h exposure amounted to 77 and 128 genes (p {<=} 0.01) at 2- and 1.8-fold filter changes, respectively. At low concentrations (0.5 and 1 mM), PB-responsive genes included the well-recognized CAR- and PXR-dependent responsive cytochromes P450 (CYP2B6, CYP3A4), sulfotransferase 2A1 and plasma transporters (ABCB1, ABCC2), as well as a number of genes critically involved in various metabolic pathways, including lipid (CYP4A11, CYP4F3), vitamin D (CYP24A1) and bile (CYP7A1 and CYP8B1) metabolism. At concentrations of 3.2 mM or higher after 20 h, and especially 48 h, increased cytotoxic effects were associated with disregulation of numerous genes related to oxidative stress, DNA repair and apoptosis. Primary human hepatocyte cultures were also exposed to 1 and 3.2 mM PB for 20 h and the changes were comparable to those found in HepaRG cells treated under the same conditions. Taken altogether, our data provide further evidence that HepaRG cells closely resemble primary human hepatocytes and provide new information on the effects of PB in human liver. These data also emphasize the importance of investigating dose- and time-dependent effects of chemicals when using toxicogenomic approaches.

  1. On the diversity of the broad emission-line profiles in active galactic nuclei

    NASA Astrophysics Data System (ADS)

    Robinson, A.

    1995-02-01

    The profiles of the broad emission lines in active galactic nuclei (AGN) vary widely in both width and shape between objects. This diversity has important consquences for models of AGN. If the conventional view that the linewidths reflect bulk velocities of the emitting gas is correct, then differences in profile shape indicate that the structure and dynamics of the broad-line region may also differ among the AGN population, even within the same phenomenological class. In this paper, the nature and possible origin of the profile diversity are discussed in terms of a simple model based on a parametrization of some basic properties of the BLR-the radial depth, line emissivity distribution and velocity law-which govern the shape of the emitted line profile. For a given geometrical configuration, the profiles produced by this model can be characterized in terms of the relative width of the core and the curvature of the wings. It is proposed that variations in both of these properties are required to account for the diversity of the Hα and Hβ profiles in a sample of Seyfert galaxies. It is not possible to identify the underlying variables unambiguously, but in this interpretation the observed profile diversity implies variations in both the slope of the line emissivity distribution and either the radial depth or the velocity law. Variations in the former two properties imply in turn that the effective size of the BLR also varies among AGN, regardless of continuum luminosity. Several consequences which follow from this are briefly discussed.

  2. Relationships Between Physical Education Students’ Motivational Profiles, Enjoyment, State Anxiety, and Self-Reported Physical Activity

    PubMed Central

    Yli-Piipari, Sami; Watt, Anthony; Jaakkola, Timo; Liukkonen, Jarmo; Nurmi, Jari-Erik

    2009-01-01

    The purpose of this study was to analyze motivational profiles based on the self-determination theory (Deci and Ryan, 2000) and how these profiles are related to physical education students’ enjoyment, state anxiety, and physical activity. The participants, 429 sixth grade students (girls = 216; boys = 213) completed SMS, Sport Enjoyment Scale, PESAS, and Physical Activity Scale. Cluster analyses identified two motivational profiles: 1) the “High motivation profile”, in which the students had high intrinsic and extrinsic motivation, and low levels of amotivation, and 2) the “Low motivation profile”, in which the students had low intrinsic and extrinsic motivation, and low levels of amotivation. The students in the first cluster enjoyed physical education more and were physically more active. The results revealed that students may be motivated towards physical education lessons both intrinsically and extrinsically, and still experience enjoyment in physical education. Key points Two motivational profiles were revealed: 1) the “High motivation profile”, in which the students had high intrinsic and extrinsic motivation, and low levels of amotivation, and 2) the “Low motivation profile”, in which the students had low intrinsic and extrinsic motivation, and low levels of amotivation. The students in the first profile enjoyed physical education more and were physically more active than the students in the second profile. Moreover, the representatives of the “High motivation profile ”experienced greater anxiety toward physical education than the representatives of the “Low motivation profile” These findings raised an interesting question whether students engaging in physical education benefit more from the presence of both self-determined and non-self-determined forms of motivation, or are the benefits higher if students are primarily self-determined? PMID:24149994

  3. Ocimum basilicum L.: phenolic profile and antioxidant-related activity.

    PubMed

    Dorman, H J Damien; Hiltunen, Raimo

    2010-01-01

    Ocimum basilicum L. leaf material was extracted by maceration with (80:20:1 v/v/v) methanol: water: acetic acid to produce a crude extract (CE), which was further fractionated by liquid-liquid extraction to isolate light petroleum (PE), ethyl acetate (EtOAc), n-butanol (n-BuOH) and H2O-soluble sub-fractions. The total phenol and flavonoid contents of the resulting samples were estimated using colorimetric-based methods, and their iron(III) reductive and free radical scavenging activities were determined in a battery of in vitro assays. The CE and sub-fractions contained phenolic compounds and flavonoids. The samples, except for PE, gave a positive result for the presence of flavones and flavonols; however, flavanones only appeared to be present in the CE. In iron(III) reduction, CE and n-BuOH were the most potent followed by EtOAc and H2O (statistically indistinguishable, p > 0.05). However, in the ferric reducing antioxidant power assay, H2O was the most potent followed by CE and EtOAc (statistically indistinguishable, p > 0.05) and n-BuOH and PE. In 1,1-diphenyl-2-picrylhydrazyl scavenging, all the samples, except PE, were effective against this reactive nitrogen species, with CE, EtOAc and n-BuOH being the most potent (statistically indistinguishable, p > 0.05). In alkylperoxyl scavenging, all the samples, except for PE, were effective against this reactive oxygen species (ROS). In superoxide anion scavenging, all the samples were capable of scavenging this ROS with CE being the most effective, followed by n-BuOH and H2O (statistically indistinguishable, p > 0.05) and EtOAc and PE. Similarly, in hydroxyl scavenging, all the samples were capable of scavenging this ROS with CE and n-BuOH being the most effective (statistically indistinguishable, p > 0.05) followed by EtOAc and H2O (statistically indistinguishable, p > 0.05) and PE.

  4. Gene expression profiling in Ishikawa cells: A fingerprint for estrogen active compounds

    SciTech Connect

    Boehme, Kathleen; Simon, Stephanie

    2009-04-01

    Several anthropogenous and naturally occurring substances, referred to as estrogen active compounds (EACs), are able to interfere with hormone and in particular estrogen receptor signaling. EACs can either cause adverse health effects in humans and wildlife populations or have beneficial effects on estrogen-dependent diseases. The aim of this study was to examine global gene expression profiles in estrogen receptor (ER)-proficient Ishikawa plus and ER-deficient Ishikawa minus endometrial cancer cells treated with selected well-known EACs (Diethylstilbestrol, Genistein, Zearalenone, Resveratrol, Bisphenol A and o,p'-DDT). We also investigated the effect of the pure antiestrogen ICI 182,780 (ICI) on the expression patterns caused by these compounds. Transcript levels were quantified 24 h after compound treatment using Illumina BeadChip Arrays. We identified 87 genes with similar expression changes in response to all EAC treatments in Ishikawa plus. ICI lowered the magnitude or reversed the expression of these genes, indicating ER dependent regulation. Apart from estrogenic gene regulation, Bisphenol A, o,p'-DDT, Zearalenone, Genistein and Resveratrol displayed similarities to ICI in their expression patterns, suggesting mixed estrogenic/antiestrogenic properties. In particular, the predominant antiestrogenic expression response of Resveratrol could be clearly distinguished from the other test compounds, indicating a distinct mechanism of action. Divergent gene expression patterns of the phytoestrogens, as well as weaker estrogenic gene expression regulation determined for the anthropogenous chemicals Bisphenol A and o,p'-DDT, warrants a careful assessment of potential detrimental and/or beneficial effects of EACs. The characteristic expression fingerprints and the identified subset of putative marker genes can be used for screening chemicals with an unknown mode of action and for predicting their potential to exert endocrine disrupting effects.

  5. Plant root transcriptome profiling reveals a strain-dependent response during Azospirillum-rice cooperation

    PubMed Central

    Drogue, Benoît; Sanguin, Hervé; Chamam, Amel; Mozar, Michael; Llauro, Christel; Panaud, Olivier; Prigent-Combaret, Claire; Picault, Nathalie; Wisniewski-Dyé, Florence

    2014-01-01

    Cooperation involving Plant Growth-Promoting Rhizobacteria results in improvements of plant growth and health. While pathogenic and symbiotic interactions are known to induce transcriptional changes for genes related to plant defense and development, little is known about the impact of phytostimulating rhizobacteria on plant gene expression. This study aims at identifying genes significantly regulated in rice roots upon Azospirillum inoculation, considering possible favored interaction between a strain and its original host cultivar. Genome-wide analyzes of Oryza sativa japonica cultivars Cigalon and Nipponbare were performed, by using microarrays, seven days post-inoculation with Azospirillum lipoferum 4B (isolated from Cigalon) or Azospirillum sp. B510 (isolated from Nipponbare) and compared to the respective non-inoculated condition. A total of 7384 genes were significantly regulated, which represent about 16% of total rice genes. A set of 34 genes is regulated by both Azospirillum strains in both cultivars, including a gene orthologous to PR10 of Brachypodium, and these could represent plant markers of Azospirillum-rice interactions. The results highlight a strain-dependent response of rice, with 83% of the differentially expressed genes being classified as combination-specific. Whatever the combination, most of the differentially expressed genes are involved in primary metabolism, transport, regulation of transcription and protein fate. When considering genes involved in response to stress and plant defense, it appears that strain B510, a strain displaying endophytic properties, leads to the repression of a wider set of genes than strain 4B. Individual genotypic variations could be the most important driving force of rice roots gene expression upon Azospirillum inoculation. Strain-dependent transcriptional changes observed for genes related to auxin and ethylene signaling highlight the complexity of hormone signaling networks in the Azospirillum

  6. Age-dependent differential expression profile of a novel intergenic long noncoding RNA in rat brain.

    PubMed

    Kour, Sukhleen; Rath, Pramod C

    2015-12-01

    Long noncoding RNAs (lncRNAs) are ≥ 200 nt long, abundant class of non-protein coding RNAs that are transcribed in complex, sense- and antisense patterns from the intergenic and intronic regions of mammalian genome. Mammalian central nervous system constitutes the largest repertoire of noncoding transcripts that are known to be expressed in developmentally regulated and cell-type specific manners. Although many lncRNAs, functioning in the brain development and diseases are known, none involved in brain aging has been reported so far. Here, we report involvement of a novel, repeat sequence (simple repeats and SINES)-containing, trans-spliced, long intergenic non-protein coding RNA (lincRNA), named as LINC-RBE (rat brain expressed transcript) involved in maturation and aging of mammalian brain. The LINC-RBE is strongly expressed in the rat brain and the upstream/downstream sequences of its DNA in the chromosome 5 contain binding sites for many cell growth, survival and development-specific transcriptional factors. Through RT-PCR and RNA in situ hybridization, LINC-RBE was found to be expressed in an age-dependent manner with significantly higher level of expression in the brain of adult (16 week) compared to both immature (4 week) and old (70 week) rats. Moreover, the expression pattern of the LINC-RBE showed distinct association with the specific neuro-anatomical regions, cell types and sub-cellular compartments of the rat brain in an age-related manner. Thus, its expression increased from immature stage to adulthood and declined further in old age. This is a first-time report of involvement of an intergenic repeat sequence-containing lncRNA in different regions of the rat brain in an age-dependent manner. Copyright © 2015 Elsevier Ltd. All rights reserved.

  7. Plant root transcriptome profiling reveals a strain-dependent response during Azospirillum-rice cooperation.

    PubMed

    Drogue, Benoît; Sanguin, Hervé; Chamam, Amel; Mozar, Michael; Llauro, Christel; Panaud, Olivier; Prigent-Combaret, Claire; Picault, Nathalie; Wisniewski-Dyé, Florence

    2014-01-01

    Cooperation involving Plant Growth-Promoting Rhizobacteria results in improvements of plant growth and health. While pathogenic and symbiotic interactions are known to induce transcriptional changes for genes related to plant defense and development, little is known about the impact of phytostimulating rhizobacteria on plant gene expression. This study aims at identifying genes significantly regulated in rice roots upon Azospirillum inoculation, considering possible favored interaction between a strain and its original host cultivar. Genome-wide analyzes of Oryza sativa japonica cultivars Cigalon and Nipponbare were performed, by using microarrays, seven days post-inoculation with Azospirillum lipoferum 4B (isolated from Cigalon) or Azospirillum sp. B510 (isolated from Nipponbare) and compared to the respective non-inoculated condition. A total of 7384 genes were significantly regulated, which represent about 16% of total rice genes. A set of 34 genes is regulated by both Azospirillum strains in both cultivars, including a gene orthologous to PR10 of Brachypodium, and these could represent plant markers of Azospirillum-rice interactions. The results highlight a strain-dependent response of rice, with 83% of the differentially expressed genes being classified as combination-specific. Whatever the combination, most of the differentially expressed genes are involved in primary metabolism, transport, regulation of transcription and protein fate. When considering genes involved in response to stress and plant defense, it appears that strain B510, a strain displaying endophytic properties, leads to the repression of a wider set of genes than strain 4B. Individual genotypic variations could be the most important driving force of rice roots gene expression upon Azospirillum inoculation. Strain-dependent transcriptional changes observed for genes related to auxin and ethylene signaling highlight the complexity of hormone signaling networks in the Azospirillum

  8. Age-dependent differential expression profile of a novel intergenic long noncoding RNA in rat brain.

    PubMed

    Kour, Sukhleen; Rath, Pramod C

    2015-11-01

    Long noncoding RNAs (lncRNAs) are ≥200 nt long, abundant class of non-protein coding RNAs that are transcribed in complex, sense- and antisense patterns from the intergenic and intronic regions of mammalian genome. Mammalian central nervous system constitutes the largest repertoire of noncoding transcripts that are known to be expressed in developmentally regulated and cell-type specific manners. Although many lncRNAs, functioning in the brain development and diseases are known, none involved in brain aging has been reported so far. Here, we report involvement of a novel, repeat sequence (simple repeats and SINES)-containing, trans-spliced, long intergenic non-protein coding RNA (lincRNA), named as LINC-RBE (rat brain expressed transcript) involved in maturation and aging of mammalian brain. The LINC-RBE is strongly expressed in the rat brain and the upstream/downstream sequences of its DNA in the chromosome 5 contain binding sites for many cell growth, survival and development-specific transcriptional factors. Through RT-PCR and RNA in situ hybridization, LINC-RBE was found to be expressed in an age-dependent manner with significantly higher level of expression in the brain of adult (16 weeks) compared to both immature (4 weeks) and old (70 weeks) rats. Moreover, the expression pattern of the LINC-RBE showed distinct association with the specific neuro-anatomical regions, cell types and sub-cellular compartments of the rat brain in an age-related manner. Thus, its expression increased from immature stage to adulthood and declined further in old age. This is a first-time report of involvement of an intergenic repeat sequence-containing lncRNA in different regions of the rat brain in an age-dependent manner.

  9. luxS-dependent gene regulation in Escherichia coli K-12 revealed by genomic expression profiling.

    PubMed

    Wang, Liang; Li, Jun; March, John C; Valdes, James J; Bentley, William E

    2005-12-01

    The bacterial quorum-sensing autoinducer 2 (AI-2) has received intense interest because the gene for its synthase, luxS, is common among a large number of bacterial species. We have identified luxS-controlled genes in Escherichia coli under two different growth conditions using DNA microarrays. Twenty-three genes were affected by luxS deletion in the presence of glucose, and 63 genes were influenced by luxS deletion in the absence of glucose. Minimal overlap among these gene sets suggests the role of luxS is condition dependent. Under the latter condition, the metE gene, the lsrACDBFG operon, and the flanking genes of the lsr operon (lsrR, lsrK, tam, and yneE) were among the most significantly induced genes by luxS. The E. coli lsr operon includes an additional gene, tam, encoding an S-adenosyl-l-methionine-dependent methyltransferase. Also, lsrR and lsrK belong to the same operon, lsrRK, which is positively regulated by the cyclic AMP receptor protein and negatively regulated by LsrR. lsrK is additionally transcribed by a promoter between lsrR and lsrK. Deletion of luxS was also shown to affect genes involved in methionine biosynthesis, methyl transfer reactions, iron uptake, and utilization of carbon. It was surprising, however, that so few genes were affected by luxS deletion in this E. coli K-12 strain under these conditions. Most of the highly induced genes are related to AI-2 production and transport. These data are consistent with the function of LuxS as an important metabolic enzyme but appear not to support the role of AI-2 as a true signal molecule for E. coli W3110 under the investigated conditions.

  10. Variations of thiaminase I activity pH dependencies among typical Great Lakes forage fish and Paenibacillus thiaminolyticus.

    USGS Publications Warehouse

    Zajicek, J.L.; Brown, L.; Brown, S.B.; Honeyfield, D.C.; Fitzsimons, J.D.; Tillitt, D.E.

    2009-01-01

    The source of thiaminase in the Great Lakes food web remains unknown. Biochemical characterization of the thiaminase I activities observed in forage fish was undertaken to provide insights into potential thiaminase sources and to optimize catalytic assay conditions. We measured the thiaminase I activities of crude extracts from five forage fish species and one strain of Paenibacillus thiaminolyticus over a range of pH values. The clupeids, alewife Alosa pseudoharengus and gizzard shad Dorosoma cepedianum, had very similar thiaminase I pH dependencies, with optimal activity ranges (> or = 90% of maximum activity) between pH 4.6 and 5.5. Rainbow smelt Osmerus mordax and spottail shiner Notropis hudsonius had optimal activity ranges between pH 5.5-6.6. The thiaminase I activity pH dependence profile of P. thiaminolyticus had an optimal activity range between pH 5.4 and 6.3, which was similar to the optimal range for rainbow smelt and spottail shiners. Incubation of P. thiaminolyticus extracts with extracts from bloater Coregonus hoyi (normally, bloaters have little or no detectable thiaminase I activity) did not significantly alter the pH dependence profile of P. thiaminolyticus-derived thiaminase I, such that it continued to resemble that of the rainbow smelt and spottail shiner, with an apparent optimal activity range between pH 5.7 and 6.6. These data are consistent with the hypothesis of a bacterial source for thiaminase I in the nonclupeid species of forage fish; however, the data also suggest different sources of thiaminase I enzymes in the clupeid species.

  11. Microbial expression profiles in the rhizosphere of willows depend on soil contamination.

    PubMed

    Yergeau, Etienne; Sanschagrin, Sylvie; Maynard, Christine; St-Arnaud, Marc; Greer, Charles W

    2014-02-01

    The goal of phytoremediation is to use plants to immobilize, extract or degrade organic and inorganic pollutants. In the case of organic contaminants, plants essentially act indirectly through the stimulation of rhizosphere microorganisms. A detailed understanding of the effect plants have on the activities of rhizosphere microorganisms could help optimize phytoremediation systems and enhance their use. In this study, willows were planted in contaminated and non-contaminated soils in a greenhouse, and the active microbial communities and the expression of functional genes in the rhizosphere and bulk soil were compared. Ion Torrent sequencing of 16S rRNA and Illumina sequencing of mRNA were performed. Genes related to carbon and amino-acid uptake and utilization were upregulated in the willow rhizosphere, providing indirect evidence of the compositional content of the root exudates. Related to this increased nutrient input, several microbial taxa showed a significant increase in activity in the rhizosphere. The extent of the rhizosphere stimulation varied markedly with soil contamination levels. The combined selective pressure of contaminants and rhizosphere resulted in higher expression of genes related to competition (antibiotic resistance and biofilm formation) in the contaminated rhizosphere. Genes related to hydrocarbon degradation were generally more expressed in contaminated soils, but the exact complement of genes induced was different for bulk and rhizosphere soils. Together, these results provide an unprecedented view of microbial gene expression in the plant rhizosphere during phytoremediation.

  12. Microbial expression profiles in the rhizosphere of willows depend on soil contamination

    PubMed Central

    Yergeau, Etienne; Sanschagrin, Sylvie; Maynard, Christine; St-Arnaud, Marc; Greer, Charles W

    2014-01-01

    The goal of phytoremediation is to use plants to immobilize, extract or degrade organic and inorganic pollutants. In the case of organic contaminants, plants essentially act indirectly through the stimulation of rhizosphere microorganisms. A detailed understanding of the effect plants have on the activities of rhizosphere microorganisms could help optimize phytoremediation systems and enhance their use. In this study, willows were planted in contaminated and non-contaminated soils in a greenhouse, and the active microbial communities and the expression of functional genes in the rhizosphere and bulk soil were compared. Ion Torrent sequencing of 16S rRNA and Illumina sequencing of mRNA were performed. Genes related to carbon and amino-acid uptake and utilization were upregulated in the willow rhizosphere, providing indirect evidence of the compositional content of the root exudates. Related to this increased nutrient input, several microbial taxa showed a significant increase in activity in the rhizosphere. The extent of the rhizosphere stimulation varied markedly with soil contamination levels. The combined selective pressure of contaminants and rhizosphere resulted in higher expression of genes related to competition (antibiotic resistance and biofilm formation) in the contaminated rhizosphere. Genes related to hydrocarbon degradation were generally more expressed in contaminated soils, but the exact complement of genes induced was different for bulk and rhizosphere soils. Together, these results provide an unprecedented view of microbial gene expression in the plant rhizosphere during phytoremediation. PMID:24067257

  13. The euryhaline yeast Debaryomyces hansenii has two catalase genes encoding enzymes with differential activity profile.

    PubMed

    Segal-Kischinevzky, Claudia; Rodarte-Murguía, Beatriz; Valdés-López, Victor; Mendoza-Hernández, Guillermo; González, Alicia; Alba-Lois, Luisa

    2011-03-01

    Debaryomyces hansenii is a spoilage yeast able to grow in a variety of ecological niches, from seawater to dairy products. Results presented in this article show that (i) D. hansenii has an inherent resistance to H2O2 which could be attributed to the fact that this yeast has a basal catalase activity which is several-fold higher than that observed in Saccharomyces cerevisiae under the same culture conditions, (ii) D. hansenii has two genes (DhCTA1 and DhCTT1) encoding two catalase isozymes with a differential enzymatic activity profile which is not strictly correlated with a differential expression profile of the encoding genes.

  14. Nocturnal activity in the green sea turtle alters daily profiles of melatonin and corticosterone.

    PubMed

    Jessop, Tim S; Limpus, Colin J; Whittier, Joan M

    2002-06-01

    In nature, green turtles (Chelonia mydas) can exhibit nocturnal activity in addition to their typically diurnal activity cycle. We examined whether nocturnal activity in captive and free-living green turtles altered daily plasma profiles of melatonin (MEL) and corticosterone (CORT). In captivity, diurnally active green turtles expressed distinct diel cycles in MEL and CORT; a nocturnal rise was observed in MEL and a diurnal rise was observed in CORT. However, when induced to perform both low- and high-intensity nocturnal activity, captive green turtles exhibited a significant decrease in MEL, compared to inactive controls. In contrast, plasma CORT increased significantly with nocturnal activity, and further, the relative increase in CORT was correlated with the intensity of the nocturnal behavior. In free-living green turtles that performed nocturnal activity including: nesting, mate searching, and feeding/swimming behaviors, plasma profiles in MEL and CORT exhibited relatively little, or no, daily fluctuation. Our findings demonstrate that nocturnal activity in green turtles is often associated with MEL and CORT profiles that resemble those measured during the day. We speculate that these conspicuous changes in MEL and CORT during nocturnal activity could either support or promote behaviors that enable acquisition of transient resources important to the survival and reproductive success of green turtles. (c) 2002 Elsevier Science (USA).

  15. Proteomic analysis of Clostridium thermocellum core metabolism: relative protein expression profiles and growth phase-dependent changes in protein expression

    PubMed Central

    2012-01-01

    Background Clostridium thermocellum produces H2 and ethanol, as well as CO2, acetate, formate, and lactate, directly from cellulosic biomass. It is therefore an attractive model for biofuel production via consolidated bioprocessing. Optimization of end-product yields and titres is crucial for making biofuel production economically feasible. Relative protein expression profiles may provide targets for metabolic engineering, while understanding changes in protein expression and metabolism in response to carbon limitation, pH, and growth phase may aid in reactor optimization. We performed shotgun 2D-HPLC-MS/MS on closed-batch cellobiose-grown exponential phase C. thermocellum cell-free extracts to determine relative protein expression profiles of core metabolic proteins involved carbohydrate utilization, energy conservation, and end-product synthesis. iTRAQ (isobaric tag for relative and absolute quantitation) based protein quantitation was used to determine changes in core metabolic proteins in response to growth phase. Results Relative abundance profiles revealed differential levels of putative enzymes capable of catalyzing parallel pathways. The majority of proteins involved in pyruvate catabolism and end-product synthesis were detected with high abundance, with the exception of aldehyde dehydrogenase, ferredoxin-dependent Ech-type [NiFe]-hydrogenase, and RNF-type NADH:ferredoxin oxidoreductase. Using 4-plex 2D-HPLC-MS/MS, 24% of the 144 core metabolism proteins detected demonstrated moderate changes in expression during transition from exponential to stationary phase. Notably, proteins involved in pyruvate synthesis decreased in stationary phase, whereas proteins involved in glycogen metabolism, pyruvate catabolism, and end-product synthesis increased in stationary phase. Several proteins that may directly dictate end-product synthesis patterns, including pyruvate:ferredoxin oxidoreductases, alcohol dehydrogenases, and a putative bifurcating hydrogenase

  16. Activation of antioxidant response element (ARE)-dependent genes by roasted coffee extracts.

    PubMed

    Yazheng, Liu; Kitts, David D

    2012-09-01

    Coffee beans contain numerous bioactive components that exhibit antioxidant capacity when assessed using both chemical, cell free, and biological, cell-based model systems. However, the mechanisms underlying the antioxidant effects of coffee in biological systems are not totally understood and in some cases vary considerably from results obtained with simpler in vitro chemical assays. In the present study, the physicochemical characteristics and antioxidant activity of roasted and non-roasted coffee extracts were investigated in both cell free (ORAC(FL)) and cell-based systems. A profile of antioxidant gene expression in cultured human colon adenocarcinoma Caco-2 cells treated with both roasted and non-roasted coffee extracts, respectively, was investigated using Real-Time polymerase chain reaction (PCR) array technology. Results demonstrated that the mechanisms of the antioxidant activity associated with coffee constituents assessed by the ORAC(FL) assay were different to those observed using an intracellular oxidation assay with Caco-2 cells. Moreover, roasted coffee (both light and dark roasted) extracts produced both increased- and decreased-expressions of numerous genes that are involved in the management of oxidative stress via the antioxidant defence system. The selective and specific positive induction of antioxidant response element (ARE)-dependent genes, including gastrointestinal glutathione peroxidase (GPX2), sulfiredoxin (SRXN1), thioredoxin reductase 1 (TXNRD1), peroxiredoxin 1 (PRDX1), peroxiredoxin 4 (PDRX4) and peroxiredoxin 6 (PDRX6) were identified with the activation of the endogenous antioxidant defence system in Caco-2 cells.

  17. Magnesium-Dependent Active-Site Conformational Selection in the Diels-Alderase Ribozyme

    SciTech Connect

    Berezniak, Tomasz; Zahran, Mai; Imhof, Petra; Jaeschke, Andres; Smith, Jeremy C

    2010-10-01

    The Diels-Alderase ribozyme, an in vitro-evolved ribonucleic acid enzyme, accelerates the formation of carbon-carbon bonds between an anthracene diene and a maleimide dienophile in a [4 + 2] cycloaddition, a reaction with broad application in organic chemistry. Here, the Diels-Alderase ribozyme is examined via molecular dynamics (MD) simulations in both crystalline and aqueous solution environments. The simulations indicate that the catalytic pocket is highly dynamic. At low Mg(2+) ion concentrations, inactive states with the catalytic pocket closed dominate. Stabilization of the enzymatically active, open state of the catalytic pocket requires a high concentration of Mg(2+) ions (e.g., 54 mM), with cations binding to specific phosphate sites on the backbone of the residues bridging the opposite strands of the pocket. The free energy profile for pocket opening at high Mg(2+) cation concentration exhibits a double minimum, with a barrier to opening of approximately 5.5 kJ/mol and the closed state approximately 3 kJ/mol lower than the open state. Selection of the open state on substrate binding leads to the catalytic activity of the ribozyme. The simulation results explain structurally the experimental observation that full catalytic activity depends on the Mg(2+) ion concentration

  18. Integrative Proteomics and Phosphoproteomics Profiling Reveals Dynamic Signaling Networks and Bioenergetics Pathways Underlying T Cell Activation.

    PubMed

    Tan, Haiyan; Yang, Kai; Li, Yuxin; Shaw, Timothy I; Wang, Yanyan; Blanco, Daniel Bastardo; Wang, Xusheng; Cho, Ji-Hoon; Wang, Hong; Rankin, Sherri; Guy, Cliff; Peng, Junmin; Chi, Hongbo

    2017-03-21

    The molecular circuits by which antigens activate quiescent T cells remain poorly understood. We combined temporal profiling of the whole proteome and phosphoproteome via multiplexed isobaric labeling proteomics technology, computational pipelines for integrating multi-omics datasets, and functional perturbation to systemically reconstruct regulatory networks underlying T cell activation. T cell receptors activated the T cell proteome and phosphoproteome with discrete kinetics, marked by early dynamics of phosphorylation and delayed ribosome biogenesis and mitochondrial activation. Systems biology analyses identified multiple functional modules, active kinases, transcription factors and connectivity between them, and mitochondrial pathways including mitoribosomes and complex IV. Genetic perturbation revealed physiological roles for mitochondrial enzyme COX10-mediated oxidative phosphorylation in T cell quiescence exit. Our multi-layer proteomics profiling, integrative network analysis, and functional studies define landscapes of the T cell proteome and phosphoproteome and reveal signaling and bioenergetics pathways that mediate lymphocyte exit from quiescence.

  19. Application of Activity-Based Protein Profiling to Study Enzyme Function in Adipocytes

    PubMed Central

    Galmozzi, Andrea; Dominguez, Eduardo; Cravatt, Benjamin F.; Saez, Enrique

    2014-01-01

    Activity-Based Protein Profiling (ABPP) is a chemical proteomics approach that utilizes small-molecule probes to determine the functional state of enzymes directly in native systems. ABPP probes selectively label active enzymes, but not their inactive forms, facilitating the characterization of changes in enzyme activity that occur without alterations in protein levels. ABPP can be a tool superior to conventional gene expression and proteomic profiling methods to discover new enzymes active in adipocytes, and to detect differences in the activity of characterized enzymes that may be associated with disorders of adipose tissue function. ABPP probes have been developed that react selectively with most members of specific enzyme classes. Here, using as an example the serine hydrolase family that includes many enzymes with critical roles in adipocyte physiology, we describe methods to apply ABPP analysis to the study of adipocyte enzymatic pathways. PMID:24529438

  20. Comprehensive Proteomic Profiling of Wheat Gluten Using a Combination of Data-Independent and Data-Dependent Acquisition

    PubMed Central

    Bromilow, Sophie N. L.; Gethings, Lee A.; Langridge, James I.; Shewry, Peter R.; Buckley, Michael; Bromley, Michael J.; Mills, E. N. Clare

    2017-01-01

    Wheat is the most important food crop in the world, the unique physiochemical properties of wheat gluten enabling a diverse range of food products to be manufactured. However, genetic and environmental factors affect the technological properties of gluten in unpredictable ways. Although newer proteomic methods have the potential to offer much greater levels of information, it is the older gel-based methods that remain most commonly used to identify compositional differences responsible for the variation in gluten functionality, in part due to the nature of their primary sequences. A combination of platforms were investigated for comprehensive gluten profiling: a QTOF with a data independent schema, which incorporated ion mobility (DIA-IM-MS) and a data dependent acquisition (DDA) workflow using a linear ion trap quadrupole (LTQ) instrument. In conjunction with a manually curated gluten sequence database a total of 2736 gluten peptides were identified with only 157 peptides identified by both platforms. These data showed 127 and 63 gluten protein accessions to be inferred with a minimum of one and three unique peptides respectively. Of the 63 rigorously identified proteins, 26 were gliadin species (4 ω-, 14 α-, and 8 γ-gliadins) and 37 glutenins (including 29 LMW glutenin and 8 HMW glutenins). Of the HMW glutenins, three were 1Dx type and five were 1Bx type illustrating the challenge of unambiguous identification of highly polymorphic proteins without cultivar specific gene sequences. The capacity of the platforms to sequence longer peptides was crucial to achieving the number of identifications, the combination of QTOF-LTQ technology being more important than extraction method to obtain a comprehensive profile. Widespread glutamine deamidation, a post-translational modification, was observed adding complexity to an already highly polymorphic mixture of proteins, with numerous insertions, deletions and substitutions. The data shown is the most comprehensive and

  1. Approaching low liver iron burden in chelated patients with non-transfusion-dependent thalassemia: the safety profile of deferasirox

    PubMed Central

    Taher, Ali T; Porter, John B; Viprakasit, Vip; Kattamis, Antonis; Chuncharunee, Suporn; Sutcharitchan, Pranee; Siritanaratkul, Noppadol; Origa, Raffaella; Karakas, Zeynep; Habr, Dany; Zhu, Zewen; Cappellini, M Domenica

    2014-01-01

    Objective Patients with non-transfusion-dependent thalassemia (NTDT) often develop iron overload and related complications, and may require iron chelation. However, the risk of over-chelation emerges as patients reach low, near-normal body iron levels and dose adjustments may be needed. In the THALASSA study, the threshold for chelation interruption was LIC <3 mg Fe/g dw (LIC<3); 24 patients receiving deferasirox for up to 2 yr reached this target. A post hoc analysis was performed to characterize the safety profile of deferasirox as these patients approached LIC<3. Methods THALASSA was a randomized, double-blind, placebo-controlled study of two deferasirox regimens (5 and 10 mg/kg/d) versus placebo in patients with NTDT. Patients randomized to deferasirox or placebo in the core could enter a 1-yr extension, with all patients receiving deferasirox (extension starting doses based on LIC at end-of-core and prior chelation response). The deferasirox safety profile was assessed between baseline and 6 months before reaching LIC<3 (Period 1), and the 6 months immediately before achieving LIC<3 (Period 2). Results Mean ± SD deferasirox treatment duration up to reaching LIC<3 was 476 ± 207 d, and deferasirox dose was 9.7 ± 3.0 mg/kg/d. The exposure-adjusted AE incidence regardless of causality was similar in periods 1 (1.026) and 2 (1.012). There were no clinically relevant differences in renal and hepatic laboratory parameters measured close to the time of LIC<3 compared with measurements near the previous LIC assessment. Conclusions The deferasirox safety profile remained consistent as patients approached the chelation interruption target, indicating that, with appropriate monitoring and dose adjustments in relation to iron load, low iron burdens may be reached with deferasirox with minimal risk of over-chelation. PMID:24460655

  2. Distinct BOLD Activation Profiles Following Central and Peripheral Oxytocin Administration in Awake Rats

    PubMed Central

    Ferris, Craig F.; Yee, Jason R.; Kenkel, William M.; Dumais, Kelly Marie; Moore, Kelsey; Veenema, Alexa H.; Kulkarni, Praveen; Perkybile, Allison M.; Carter, C. Sue

    2015-01-01

    A growing body of literature has suggested that intranasal oxytocin (OT) or other systemic routes of administration can alter prosocial behavior, presumably by directly activating OT sensitive neural circuits in the brain. Yet there is no clear evidence that OT given peripherally can cross the blood–brain barrier at levels sufficient to engage the OT receptor. To address this issue we examined changes in blood oxygen level-dependent (BOLD) signal intensity in response to peripheral OT injections (0.1, 0.5, or 2.5 mg/kg) during functional magnetic resonance imaging (fMRI) in awake rats imaged at 7.0 T. These data were compared to OT (1 μg/5 μl) given directly to the brain via the lateral cerebroventricle. Using a 3D annotated MRI atlas of the rat brain segmented into 171 brain areas and computational analysis, we reconstructed the distributed integrated neural circuits identified with BOLD fMRI following central and peripheral OT. Both routes of administration caused significant changes in BOLD signal within the first 10 min of administration. As expected, central OT activated a majority of brain areas known to express a high density of OT receptors, e.g., lateral septum, subiculum, shell of the accumbens, bed nucleus of the stria terminalis. This profile of activation was not matched by peripheral OT. The change in BOLD signal to peripheral OT did not show any discernible dose–response. Interestingly, peripheral OT affected all subdivisions of the olfactory bulb, in addition to the cerebellum and several brainstem areas relevant to the autonomic nervous system, including the solitary tract nucleus. The results from this imaging study do not support a direct central action of peripheral OT on the brain. Instead, the patterns of brain activity suggest that peripheral OT may interact at the level of the olfactory bulb and through sensory afferents from the autonomic nervous system to influence brain activity. PMID:26441574

  3. Comparison of parameters of bone profile and homocysteine in physically active and non-active postmenopausal females

    PubMed Central

    Tariq, Sundus; Lone, Khalid Parvez; Tariq, Saba

    2016-01-01

    Background and objectives: Optimal physical activity is important in attaining a peak bone mass. Physically active women have better bone mineral density and reduce fracture risk as compared to females living a sedentary life. The objective of this study was to compare parameters of bone profile and serum homocysteine levels in physically active and non-active postmenopausal females. Methods: In this cross sectional study postmenopausal females between 50-70 years of age were recruited and divided into two groups: Physically inactive (n=133) performing light physical activity and Physically active (n=34) performing moderate physical activity. Physical activity (in metabolic equivalents), bone mineral density and serum homocysteine levels were assessed. Spearman’s rho correlation was applied to observe correlations. Two independent sample t test and Mann Whitney U test were applied to compare groups. P-value ≤ 0.05 was taken statistically significant. Results: Parameters of bone profile were significantly higher and serum homocysteine levels were significantly lower in postmenopausal females performing moderate physical activity as compared to females performing light physical activity. Homocysteine was not significantly related to T-score and Z-score in both groups. Conclusion: Improving physical activity could be beneficial for improving the quality of bone, decreasing fracture risk and decreasing serum homocysteine levels. PMID:27882033

  4. Fyn phosphorylates AMPK to inhibit AMPK activity and AMP-dependent activation of autophagy

    PubMed Central

    Yamada, Eijiro; Okada, Shuichi; Bastie, Claire C.; Vatish, Manu; Nakajima, Yasuyo; Shibusawa, Ryo; Ozawa, Atsushi; Pessin, Jeffrey E.; Yamada, Masanobu

    2016-01-01

    We previously demonstrated that proto-oncogene Fyn decreased energy expenditure and increased metabolic phenotypes. Also Fyn decreased autophagy-mediated muscle mass by directly inhibiting LKB1 and stimulating STAT3 activities, respectively. AMPK, a downstream target of LKB1, was recently identified as a key molecule controlling autophagy. Here we identified that Fyn phosphorylates the α subunit of AMPK on Y436 and inhibits AMPK enzymatic activity without altering the assembly state of the AMPK heterotrimeric complex. As pro-inflammatory mediators are reported modulators of the autophagy processes, treatment with the pro-inflammatory cytokine TNFα resulted in 1) increased Fyn activity 2) stimulated Fyn-dependent AMPKα tyrosine phosphorylation and 3) decreased AICAR-dependent AMPK activation. Importantly, TNFα induced inhibition of autophagy was not observed when AMPKα was mutated on Y436. 4) These data demonstrate that Fyn plays an important role in relaying the effects of TNFα on autophagy and apoptosis via phosphorylation and inhibition of AMPK. PMID:27626315

  5. Hα Surface Brightness Profiles of Star-Forming Galaxies and Dependence on Halo Mass Using the HAGGIS Survey

    NASA Astrophysics Data System (ADS)

    Kulkarni, S.; Wilman, D.; Erwin, P.; Koppenhöfer, J.; Gutierrez, L.; Beckman, J.; Saglia, R.; Bender, R.

    2014-03-01

    We present the first results from the Hα Galaxy Groups Imaging Survey (HAGGIS), a narrow-band imaging survey of SDSS groups at z < 0.05 conducted using the Wide Field Imager (WFI) on the ESO/MPG 2.2-meter telescope and the Wide Field Camera (WFC) on the Issac Newton Telescope (INT). In total, we observed 100 galaxy groups with a wide range of halo mass (1012 - 1014 M⊙) in pairs of narrow-band filters selected to get continuum subtracted rest-frame Hα images for each galaxy. The excellent data allows us to detect Hα down to the 10-18 ergs/s/cm2/arcsec2 level. Here, we examine the role played by halo mass and galaxy stellar mass in deciding the overall star formation activity in star forming disks by comparing stacked Hα profiles of galaxies in different halo mass and stellar mass bins. With this preliminary study, we have found that the star-formation activity in star-forming galaxies decreases in larger halos compared to the field galaxies. Using median equivalent width profiles, we can infer how environmental processes affect star-forming galaxies differently at different radii.

  6. Gene Expression Profile of Calcium/Calmodulin-Dependent Protein Kinase IIα in Rat's Hippocampus during Morphine Withdrawal

    PubMed Central

    Ahmadi, Shamseddin; Amiri, Shahin; Rafieenia, Fatemeh; Rostamzadeh, Jalal

    2013-01-01

    Introduction Calcium/calmodulin-dependent protein kinase II (CaMKII) which is highly expressed in the hippocampus is known to play a pivotal role in reward-related memories and morphine dependence. Methods In the present study, repeated morphine injections once daily for 7 days was done to induce morphine tolerance in male Wistar rats, after which gene expression profile of α-isoform of CaMKII (CaMKIIα) in the hippocampus was evaluated upon discontinuation of morphine injection over 21 days of morphine withdrawal. Control groups received saline for 7 consecutive days. For gene expression study, rats’ brains were removed and the hippocampus was dissected in separate groups on days 1, 3, 7, 14, and 21 since discontinuation of of morphine injection. A semi-quantitative RT-PCR method was used to evaluate the gene expression profile. Results Tolerance to morphine was verified by a significant decrease in morphine analgesia in a hotplate test on day 8 (one day after the final repeated morphine injections). Results showed that gene expression of CaMKIIα at mRNA level on day 1, 3, 7, 14 and 21 of morphine withdrawal was significantly altered as compared to the saline control group. Post hoc Tukey's test revealed a significantly enhanced CaMKIIα gene expression on day 14. Discussion It can be concluded that CaMKIIα gene expression during repeated injections of morphine is increased and this increase continues up to 14 days of withdrawal then settles at a new set point. Therefore, the strong morphine reward-related memory in morphine abstinent animals may, at least partly be attributed to, the up-regulation of CaMKIIα in the hippocampus over 14 days of morphine withdrawal. PMID:25337341

  7. Profiling physical activity motivation based on self-determination theory: a cluster analysis approach.

    PubMed

    Friederichs, Stijn Ah; Bolman, Catherine; Oenema, Anke; Lechner, Lilian

    2015-01-01

    In order to promote physical activity uptake and maintenance in individuals who do not comply with physical activity guidelines, it is important to increase our understanding of physical activity motivation among this group. The present study aimed to examine motivational profiles in a large sample of adults who do not comply with physical activity guidelines. The sample for this study consisted of 2473 individuals (31.4% male; age 44.6 ± 12.9). In order to generate motivational profiles based on motivational regulation, a cluster analysis was conducted. One-way analyses of variance were then used to compare the clusters in terms of demographics, physical activity level, motivation to be active and subjective experience while being active. Three motivational clusters were derived based on motivational regulation scores: a low motivation cluster, a controlled motivation cluster and an autonomous motivation cluster. These clusters differed significantly from each other with respect to physical activity behavior, motivation to be active and subjective experience while being active. Overall, the autonomous motivation cluster displayed more favorable characteristics compared to the other two clusters. The results of this study provide additional support for the importance of autonomous motivation in the context of physical activity behavior. The three derived clusters may be relevant in the context of physical activity interventions as individuals within the different clusters might benefit most from different intervention approaches. In addition, this study shows that cluster analysis is a useful method for differentiating between motivational profiles in large groups of individuals who do not comply with physical activity guidelines.

  8. Cell Expansion-Dependent Inflammatory and Metabolic Profile of Human Bone Marrow Mesenchymal Stem Cells

    PubMed Central

    Prieto, Patricia; Fernández-Velasco, María; Fernández-Santos, María E.; Sánchez, Pedro L.; Terrón, Verónica; Martín-Sanz, Paloma; Fernández-Avilés, Francisco; Boscá, Lisardo

    2016-01-01

    Stem cell therapy has emerged as a promising new area in regenerative medicine allowing the recovery of viable tissues. Among the many sources of adult stem cells, bone marrow-derived are easy to expand in culture via plastic adherence and their multipotentiality for differentiation make them ideal for clinical applications. Interestingly, several studies have indicated that MSCs expansion in vitro may be limited mainly due to “cell aging” related to the number of cell divisions in culture. We have determined that MSCs exhibit a progressive decline across successive passages in the expression of stem cell markers, in plasticity and in the inflammatory response, presenting low immunogenicity. We have exposed human MSCs after several passages to TLRs ligands and analyzed their inflammatory response. These cells responded to pro-inflammatory stimuli (i.e., NOS-2 expression) and to anti-inflammatory cytokines (i.e., HO1 and Arg1) until two expansions, rapidly declining upon subculture. Moreover, in the first passages, MSCs were capable to release IL1β, IL6, and IL8, as well as to produce active MMPs allowing them to migrate. Interestingly enough, after two passages, anaerobic glycolysis was enhanced releasing high levels of lactate to the extracellular medium. All these results may have important implications for the safety and efficacy of MSCs-based cell therapies. PMID:27899899

  9. Continuous Water Vapor Profiles from Operational Ground-Based Active and Passive Remote Sensors

    NASA Technical Reports Server (NTRS)

    Turner, D. D.; Feltz, W. F.; Ferrare, R. A.

    2000-01-01

    The Atmospheric Radiation Measurement program's Southern Great Plains Cloud and Radiation Testbed site central facility near Lamont, Oklahoma, offers unique operational water vapor profiling capabilities, including active and passive remote sensors as well as traditional in situ radiosonde measurements. Remote sensing technologies include an automated Raman lidar and an automated Atmospheric Emitted Radiance Interferometer (AERI), which are able to retrieve water vapor profiles operationally through the lower troposphere throughout the diurnal cycle. Comparisons of these two water vapor remote sensing methods to each other and to radiosondes over an 8-month period are presented and discussed, highlighting the accuracy and limitations of each method. Additionally, the AERI is able to retrieve profiles of temperature while the Raman lidar is able to retrieve aerosol extinction profiles operationally. These data, coupled with hourly wind profiles from a 915-MHz wind profiler, provide complete specification of the state of the atmosphere in noncloudy skies. Several case studies illustrate the utility of these high temporal resolution measurements in the characterization of mesoscale features within a 3-day time period in which passage of a dryline, warm air advection, and cold front occurred.

  10. Automated Structure-Activity Relationship Mining: Connecting Chemical Structure to Biological Profiles.

    PubMed

    Wawer, Mathias J; Jaramillo, David E; Dančík, Vlado; Fass, Daniel M; Haggarty, Stephen J; Shamji, Alykhan F; Wagner, Bridget K; Schreiber, Stuart L; Clemons, Paul A

    2014-06-01

    Understanding the structure-activity relationships (SARs) of small molecules is important for developing probes and novel therapeutic agents in chemical biology and drug discovery. Increasingly, multiplexed small-molecule profiling assays allow simultaneous measurement of many biological response parameters for the same compound (e.g., expression levels for many genes or binding constants against many proteins). Although such methods promise to capture SARs with high granularity, few computational methods are available to support SAR analyses of high-dimensional compound activity profiles. Many of these methods are not generally applicable or reduce the activity space to scalar summary statistics before establishing SARs. In this article, we present a versatile computational method that automatically extracts interpretable SAR rules from high-dimensional profiling data. The rules connect chemical structural features of compounds to patterns in their biological activity profiles. We applied our method to data from novel cell-based gene-expression and imaging assays collected on more than 30,000 small molecules. Based on the rules identified for this data set, we prioritized groups of compounds for further study, including a novel set of putative histone deacetylase inhibitors. © 2014 Society for Laboratory Automation and Screening.

  11. Automated structure–activity relationship mining: connecting chemical structure to biological profiles

    PubMed Central

    Wawer, Mathias J.; Jaramillo, David E.; Dancik, Vlado; Fass, Daniel M.; Haggarty, Stephen J.; Shamji, Alykhan F.; Wagner, Bridget K.; Schreiber, Stuart L.; Clemons, Paul A.

    2017-01-01

    Understanding structure–activity relationships (SARs) of small molecules is important for developing probes and novel therapeutic agents in chemical biology and drug discovery. Increasingly multiplexed small-molecule profiling assays allow simultaneous measurement of many biological response parameters for the same compound, e.g. expression levels for many genes or binding constants against many proteins. While such methods promise to capture SARs with high granularity, few computational methods are available to support SAR analyses of high-dimensional compound activity profiles. Many of these methods are not generally applicable or reduce the activity space to scalar summary statistics before establishing SARs. In this article, we present a versatile computational method that automatically extracts interpretable SAR rules from high-dimensional profiling data. The rules connect chemical structural features of compounds to patterns in their biological activity profiles. We applied our method to data from novel cell-based gene-expression and imaging assays collected on more than 30,000 small molecules. Based on the rules identified for this dataset, we prioritized groups of compounds for further study, including a novel set of putative histone deacetylase inhibitors. PMID:24710340

  12. NESmapper: accurate prediction of leucine-rich nuclear export signals using activity-based profiles.

    PubMed

    Kosugi, Shunichi; Yanagawa, Hiroshi; Terauchi, Ryohei; Tabata, Satoshi

    2014-09-01

    The nuclear export of proteins is regulated largely through the exportin/CRM1 pathway, which involves the specific recognition of leucine-rich nuclear export signals (NESs) in the cargo proteins, and modulates nuclear-cytoplasmic protein shuttling by antagonizing the nuclear import activity mediated by importins and the nuclear import signal (NLS). Although the prediction of NESs can help to define proteins that undergo regulated nuclear export, current methods of predicting NESs, including computational tools and consensus-sequence-based searches, have limited accuracy, especially in terms of their specificity. We found that each residue within an NES largely contributes independently and additively to the entire nuclear export activity. We created activity-based profiles of all classes of NESs with a comprehensive mutational analysis in mammalian cells. The profiles highlight a number of specific activity-affecting residues not only at the conserved hydrophobic positions but also in the linker and flanking regions. We then developed a computational tool, NESmapper, to predict NESs by using profiles that had been further optimized by training and combining the amino acid properties of the NES-flanking regions. This tool successfully reduced the considerable number of false positives, and the overall prediction accuracy was higher than that of other methods, including NESsential and Wregex. This profile-based prediction strategy is a reliable way to identify functional protein motifs. NESmapper is available at http://sourceforge.net/projects/nesmapper.

  13. HPLC profiling and quantification of active principles in leaves of Hedera helix L.

    PubMed

    Demirci, B; Goppel, M; Demirci, F; Franz, G

    2004-10-01

    Ivy (Hedera helix L., Araliaceae), is an evergreen medicinal and ornamental plant. Depending on leaf polymorphism different shaped ivy leaves were extracted and subsequently analyzed by reversed-phase high performance liquid chromatography (RP-HPLC). Quantitative determination of its most prominent saponins hederacoside C (1) and alpha-hederin (2) from different ivy leaf extracts were detected, validated and optimized for quick profiling. The linearity of response, repeatability and reproducibility of the applied RP-HPLC method are reported.

  14. Do Training Programs Work? An Assessment of Pharmacists Activities in the Field of Chemical Dependency.

    ERIC Educational Resources Information Center

    Brooks, Valerie G.; Brock, Tina Penick; Ahn, Jungeun

    2001-01-01

    Seeks to determine if pharmacists who attended a chemical dependency training program were performing more chemical dependency related activities. Results reveal that participants were more likely to perform the following activities: lecture to community groups about chemical dependency; participate in a pharmacists' recovery program; provide…

  15. Ischemia Induced Neutrophil Activation and Diapedesis is Lipoxygenase Dependent.

    DTIC Science & Technology

    2007-11-02

    activity in mediating PMN activation and diapedesis . Anesthetized rabbits (n = 8) underwent 3 h of bilateral hindlimb ischemia. At 10 min of reperfusion...enhanced response of 337% to PMA stimulation. To study diapedesis , plasma collected at 10 min of reperfusion was introduced into plastic chambers taped...abolished PMN activation (51 +/- 12 fM DCF/cell) and ischemic plasma induced diapedesis into the plastic chamber (38 +/- 18 PMN/mm(exp 3)).

  16. The interaction between physical activity and fasting on the serum lipid profile during Ramadan.

    PubMed

    Haghdoost, A A; Poorranjbar, M

    2009-09-01

    The serum lipid profiles in Muslims change during the fasting month of Ramadan, but it is not clear whether this change is due to changes in their physical activities. In this study, we compared the patterns of the lipid profile changes in those who engaged in regular physical activity with those who did not. In a randomised trial, we assigned 93 students who took a physical education course into two groups--those who had regular physical activity after Ramadan and those who had physical activity during Ramadan. Venous blood (5 ml) was taken just before, at the end, and 40 days after Ramadan, and the fasting glucose sugar and lipid profile were measured. Fasting with physical activity decreased body weight by 1.2 kg (p-value is 0.03). Fasting blood sugar also decreased by 7 mg/dL during Ramadan, but this drop was observed in both groups. Triglyceride decreased in both groups during Ramadan, but cholesterol levels dropped considerably during and after Ramadan for those who concurrently engaged in physical activity and fasted (-12.24 and -8.4 mg/dL, respectively). The patterns of changes in the high-density lipoprotein (HDL), low-density lipoprotein (LDL) and HDL/LDL values were more or less comparable in both groups (p is greater than 0.5). Usually, people are less physically active during Ramadan, but our findings show that physical activity alone cannot explain the variations in the lipid profile. Other factors, such as changes in the diet and sleeping hours, may have more important roles.

  17. Association of physical activity and physical fitness with blood pressure profile in Gujarati Indian adolescents.

    PubMed

    Shaikh, Wasim A; Patel, Minal C; Singh, S K

    2011-01-01

    The current study was conducted to determine how physical activity level and physical fitness affects the blood pressure profile of Gujarati Indian adolescents so as to help in developing preventive strategies for the local population as ethnic differences exist in the aetiopathogenesis of hypertension. A cross-sectional study was conducted on 485 Gujarati Indian adolescent boys and girls of age group 16-19 years. Physical activity level was assessed using Johnson Space Center/NASA Physical Activity Rating Scale and VO2 max was used to assess the physical fitness. Body composition was assessed in terms of Body Mass Index, Fat Mass Index and Waist Circumference. Blood Pressure was measured by oscillometry. One-way ANOVA was used to study if any significant differences (P<0.05) existed in the blood pressure profile between the high, moderate and low physical activity groups. Pearson's correlation coefficient was determined to assess the relationship between VO2 max and blood pressure profile. In girls, physical activity level was not found to have a significant effect on the blood pressure profile. In boys, systolic blood pressure and mean arterial pressure were found to be significantly higher in Moderate Physical Activity Group as compared to Low Physical Activity Group. PVO2 max was found to have a significant negative correlationship with SBP, DBP and MAP in girls and a significant negative correlationship with SBP, PP and MAP in boys. It could thus be concluded that a better physical fitness rather than a higher physical activity level could keep the blood pressure in check in the Gujarati Indian adolescents.

  18. Spatially dependent activation of the patterning protease, Easter.

    PubMed

    LeMosy, Ellen K

    2006-04-17

    The dorsoventral axis of the Drosophila embryo is established by the activating cleavage of a signaling ligand by a serine protease, Easter, only on the ventral side of the embryo. Easter is the final protease in a serine protease cascade in which initial reaction steps appear not to be ventrally restricted, but where Easter activity is promoted ventrally through the action of a spatial cue at an unknown step in the pathway. Here, biochemical studies demonstrate that this spatial control occurs at or above the level of Easter zymogen activation, rather than through direct promotion of Easter's catalytic activity against the signaling ligand.

  19. Adolescents' physical activity in physical education, school recess, and extra-curricular sport by motivational profiles.

    PubMed

    Mayorga-Vega, Daniel; Viciana, Jesús

    2014-06-01

    The main purpose of this study was to evaluate the differences in adolescents´ objective physical activity levels and perceived effort in physical education, school recess, and extra-curricular organized sport by motivational profiles in physical education. A sample of 102 students 11-16 yr. old completed a self-report questionnaire assessing self-determined motivation toward physical education. Subsequently, students' objective physical activity levels (steps/min., METs, and moderate-to-vigorous physical activity) and perceived effort were evaluated for each situation. Cluster analysis identified a two-cluster structure: "Moderate motivation toward physical education profile" and "High motivation toward physical education profile." Adolescents in the second cluster had higher physical activity and perceived effort values than adolescents in the first cluster, except for METs and moderate-to-vigorous physical activity in extra-curricular sport. These results support the importance of physical education teachers who should promote self-determined motivation toward physical education so that students can reach the recommended physical activity levels.

  20. Assessing microbial communities for a metabolic profile similar to activated sludge.

    PubMed

    Paixão, S M; Sàágua, M C; Tenreiro, R; Anselmo, A M

    2007-05-01

    To search for reliable testing inocula alternatives to activated sludge cultures, several model microbial consortia were compared with activated sludge populations for their functional diversity. The evaluation of the metabolic potential of these mixed inocula was performed using the Biolog EcoPlates and GN and GP MicroPlates (Biolog, Inc., Hayward, California). The community-level physiological profiles (CLPPs) obtained for model communities and activated sludge samples were analyzed by principal component analysis and hierarchic clustering methods, to evaluate the ability of Biolog plates to distinguish among the different microbial communities. The effect of different inocula preparation methodologies on the community structure was also studied. The CLPPs obtained with EcoPlates and GN MicroPlates showed that EcoPlates are suitable to screen communities with a metabolic profile similar to activated sludge. New, well-defined, standardized, and safe inocula presenting the same metabolic community profile as activated sludge were selected and can be tested as surrogate cultures in activated-sludge-based bioassays.

  1. Field dependence-independence and participation in physical activity by college students.

    PubMed

    Liu, Wenhao

    2006-06-01

    Field-independent individuals, compared with field-dependent individuals, have higher sports potential and advantages in sport-related settings. Little research, however, has been conducted on the association of field dependence-independence and participation in physical activity. The study examined this association for college students who participated in physical activities in and beyond physical education classes. The Group Embedded Figures Test distinguished 40 field-dependent from 40 field-independent participants. Activity logs during one semester showed that field-independent participants were significantly more physically active and their physical activity behaviors were more sport-related than those of field-dependent participants.

  2. Cytotoxic activity in children with insulin-dependent diabetes mellitus.

    PubMed

    Lorini, R; Moretta, A; Valtorta, A; d'Annunzio, G; Cortona, L; Vitali, L; Bozzola, M; Severi, F

    1994-02-01

    We determined the percentage of circulating natural killer (NK) cells, using the monoclonal antibodies anti-CD57 and anti-CD16, NK cytotoxic activity (lytic units/10(6)) and lymphokine-activated killer (LAK) activity in 25 IDDM patients aged 3-23 years, 12 with disease for < 1 year (Group I) and 13 with disease for > 3 years (Group II). Nine age-matched healthy subjects served as controls. The percentage of CD57+ cells was similar in IDDM patients and controls, while the percentage of CD16+ cells was lower in IDDM patients (P < 0.05) than in controls. NK cell cytotoxic activity was lower in IDDM patients than in controls (P < 0.01), in Group I and II compared with controls (P < 0.005). LAK activity was similar in IDDM patients and in controls. No correlation was found between NK cytotoxic activity and metabolic control, HLA typing, while a negative correlation was found between NK cytotoxic activity and insulin requirement (P < 0.05). The decreased NK cytotoxic activity observed in our patients, in particular in long-standing diabetics, with normal NK cell number, could be due to a qualitative defect of the NK cells, or to a deficient IL-2 and/or TNF-alpha production, or to a immunomodulatory or immunosuppressing effect of insulin.

  3. Time-Dependent Expression Profiles of microRNAs and mRNAs in Rat Milk Whey

    PubMed Central

    Izumi, Hirohisa; Kosaka, Nobuyoshi; Shimizu, Takashi; Sekine, Kazunori; Ochiya, Takahiro; Takase, Mitsunori

    2014-01-01

    Functional RNAs, such as microRNA (miRNA) and mRNA, are present in milk, but their roles are unknown. To clarify the roles of milk RNAs, further studies using experimental animals such as rats are needed. However, it is unclear whether rat milk also contains functional RNAs and what their time dependent expression profiles are. Thus, we prepared total RNA from whey isolated from rat milk collected on days 2, 9, and 16 postpartum and analyzed using microarrays and quantitative PCR. The concentration of RNA in colostrum whey (day 2) was markedly higher than that in mature milk whey (days 9 and 16). Microarray analysis detected 161 miRNAs and 10,948 mRNA transcripts. Most of the miRNAs and mRNA transcripts were common to all tested milks. Finally, we selected some immune- and development-related miRNAs and mRNAs, and analysed them by quantitative PCR (in equal sample volumes) to determine their time-dependent changes in expression in detail. Some were significantly more highly expressed in colostrum whey than in mature milk whey, but some were expressed equally. And mRNA expression levels of some cytokines and hormones did not reflect the protein levels. It is still unknown whether RNAs in milk play biological roles in neonates. However, our data will help guide future in vivo studies using experimental animals such as rats. PMID:24533154

  4. Tumour-site-dependent expression profile of angiogenic factors in tumour-associated stroma of primary colorectal cancer and metastases.

    PubMed

    Kahlert, C; Pecqueux, M; Halama, N; Dienemann, H; Muley, T; Pfannschmidt, J; Lasitschka, F; Klupp, F; Schmidt, T; Rahbari, N; Reissfelder, C; Kunz, C; Benner, A; Falk, C; Weitz, J; Koch, M

    2014-01-21

    Tumour-associated stroma has a critical role in tumour proliferation. Our aim was to determine a specific protein expression profile of stromal angiogenic cytokines and matrix metalloproteinases (MMPs) to identify potential biomarkers or new therapy targets. Frozen tissue of primary colorectal cancer (n=25), liver (n=25) and lung metastases (n=23) was laser-microdissected to obtain tumour epithelial cells and adjacent tumour-associated stroma. Protein expression of nine angiogenic cytokines and eight MMPs was analysed using a multiplex-based protein assay. We found a differential expression of several MMPs and angiogenic cytokines in tumour cells compared with adjacent tumour stroma. Cluster analysis displayed a tumour-site-dependent stromal expression of MMPs and angiogenic cytokines. Univariate analysis identified stromal MMP-2 and MMP-3 in primary colorectal cancer, stromal MMP-1, -2, -3 and Angiopoietin-2 in lung metastases and stromal MMP-12 and VEGF in liver metastases as prognostic markers (P>0.05, respectively). Furthermore, stroma-derived Angiopoietin-2 proved to be an independent prognostic marker in colorectal lung metastases. Expression of MMPs and angiogenic cytokines in tumour cells and adjacent tumour stroma is dependent on the tumour site. Stroma-derived MMPs and angiogenic cytokines may be useful prognostic biomarkers. These data can be helpful to identify new agents for a targeted therapy in patients with colorectal cancer.

  5. Profiles of recreational activities of daily living (RADL) in patients with mental disorders.

    PubMed

    Linden, Michael; Gehrke, G; Geiselmann, B

    2009-12-01

    Activities of daily living, play a key role in the measurement of functional health as defined by the International Classification of Functioning, Disability and Health (ICF) and in prevention and treatment of mental or somatic illnesses. From a clinical context it is important to discriminate between basic "activities of daily living, ADL", "intentional activities of daily living, IADL", and "recreational activities of daily living, RADL". While ADL and IADL have gained much attention in dementia, the elderly, or severe somatic illnesses, there is a lack of research on RADL, which are important in depression, anxiety, or other neurotic disorders. 154 unselected inpatients of a department of behavioral and psychosomatic medicine filled in the "Check List of Recreational Activities" to assess the rates and profiles of RADL. Patients reported on average 19.3 (s.d. 7.0) activities (range 4 - 40), i.e. males 21.3 (s.d. 6.5, 9 - 34) and females 18.9 (s.d. 7.1, 4 - 40). Most frequent RADL were passive and unspecific activities like "watching tv" (93.4%). Least frequent were activities which need special skills or preparation like "horse back riding" (0.7%). Low rates were also found for activities which are in the centre of inpatient occupational therapy like "ceramics" (4.7%) or "silk-painting" (2.6%). There are differences between sexes but not in respect to age (18 to 60), sick leave and unemployment, or diagnostic status. When patients were asked what they would like to do in the future, the same activity profile emerged as when looking at what they had done in the last month The data give a reference profile for recreational activities, help to define what can be considered a normal frequency and spectrum of RADL, and, by this, can guide therapeutic interventions.

  6. Effects of gamma irradiation on chickpea seeds vis-a-vis total seed storage proteins, antioxidant activity and protein profiling.

    PubMed

    Bhagyawant, S S; Gupta, N; Shrivastava, N

    2015-10-23

    The present work describes radiation—induced effects on seed composition vis—à—vis total seed proteins, antioxidant levels and protein profiling employing two dimensional gel electrophoresis (2D—GE) in kabuli and desi chickpea varities. Seeds were exposed to the radiation doses of 1,2,3,4 and 5 kGy. The total protein concentrations decreased and antioxidant levels were increased with increasing dose compared to control seed samples. Radiation induced effects were dose dependent to these seed parameters while it showed tolerance to 1 kGy dose. Increase in the dose was complimented with increase in antioxidant levels, like 5 kGy enhanced % scavenging activities in all the seed extracts. Precisely, the investigations reflected that the dose range from 2 to 5 kGy was effective for total seed storage proteins, as depicted quantitatively and qualitative 2D—GE means enhance antioxidant activities in vitro.

  7. Autoregressive Higher-Order Hidden Markov Models: Exploiting Local Chromosomal Dependencies in the Analysis of Tumor Expression Profiles

    PubMed Central

    Seifert, Michael; Abou-El-Ardat, Khalil; Friedrich, Betty; Klink, Barbara; Deutsch, Andreas

    2014-01-01

    Changes in gene expression programs play a central role in cancer. Chromosomal aberrations such as deletions, duplications and translocations of DNA segments can lead to highly significant positive correlations of gene expression levels of neighboring genes. This should be utilized to improve the analysis of tumor expression profiles. Here, we develop a novel model class of autoregressive higher-order Hidden Markov Models (HMMs) that carefully exploit local data-dependent chr