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Sample records for raphe terminus creating

  1. Absence of lateral palpebral raphe in Caucasians

    Directory of Open Access Journals (Sweden)

    Lucy Goold

    2009-07-01

    Full Text Available Lucy Goold1, Hirohiko Kakizaki1,2,3, Raman Malhotra3, Dinesh Selva11South Australian Institute of Ophthalmology and Discipline of Ophthalmology and Visual Sciences, University of Adelaide, Australia; 2Department of Ophthalmology, Aichi Medical University, Nagakute, Aichi, Japan; 3Corneoplastic Unit and Eye Bank, Queen Victoria Hospital NHS Trust, East Grinstead, West Sussex, United KingdomAbstract: Classical anatomical teaching reports the presence of the lateral palpebral raphe formed at the union in the preseptal and orbital parts of the orbicularis oculi muscle, or by the tendon adhering these to the underlying zygomatic bone. The lateral palpebral raphe has been shown to be absent in Asian cadavers. The current study uses both evidence from the anatomical dissection of five eyelids from three Caucasian cadavers, and histological assessment of the lateral canthus of 13 eyelids from seven Caucasian cadavers to illustrate the absence of the lateral palpebral raphe in Caucasian population.Keywords: lateral palpebral raphe, orbicularis oculi muscle, Caucasian, cadavers

  2. Activity of Raphé Serotonergic Neurons Controls Emotional Behaviors

    Directory of Open Access Journals (Sweden)

    Anne Teissier

    2015-12-01

    Full Text Available Despite the well-established role of serotonin signaling in mood regulation, causal relationships between serotonergic neuronal activity and behavior remain poorly understood. Using a pharmacogenetic approach, we find that selectively increasing serotonergic neuronal activity in wild-type mice is anxiogenic and reduces floating in the forced-swim test, whereas inhibition has no effect on the same measures. In a developmental mouse model of altered emotional behavior, increased anxiety and depression-like behaviors correlate with reduced dorsal raphé and increased median raphé serotonergic activity. These mice display blunted responses to serotonergic stimulation and behavioral rescues through serotonergic inhibition. Furthermore, we identify opposing consequences of dorsal versus median raphé serotonergic neuron inhibition on floating behavior, together suggesting that median raphé hyperactivity increases anxiety, whereas a low dorsal/median raphé serotonergic activity ratio increases depression-like behavior. Thus, we find a critical role of serotonergic neuronal activity in emotional regulation and uncover opposing roles of median and dorsal raphé function.

  3. Activity of Raphé Serotonergic Neurons Controls Emotional Behaviors.

    Science.gov (United States)

    Teissier, Anne; Chemiakine, Alexei; Inbar, Benjamin; Bagchi, Sneha; Ray, Russell S; Palmiter, Richard D; Dymecki, Susan M; Moore, Holly; Ansorge, Mark S

    2015-12-01

    Despite the well-established role of serotonin signaling in mood regulation, causal relationships between serotonergic neuronal activity and behavior remain poorly understood. Using a pharmacogenetic approach, we find that selectively increasing serotonergic neuronal activity in wild-type mice is anxiogenic and reduces floating in the forced-swim test, whereas inhibition has no effect on the same measures. In a developmental mouse model of altered emotional behavior, increased anxiety and depression-like behaviors correlate with reduced dorsal raphé and increased median raphé serotonergic activity. These mice display blunted responses to serotonergic stimulation and behavioral rescues through serotonergic inhibition. Furthermore, we identify opposing consequences of dorsal versus median raphé serotonergic neuron inhibition on floating behavior, together suggesting that median raphé hyperactivity increases anxiety, whereas a low dorsal/median raphé serotonergic activity ratio increases depression-like behavior. Thus, we find a critical role of serotonergic neuronal activity in emotional regulation and uncover opposing roles of median and dorsal raphé function. Copyright © 2015 The Authors. Published by Elsevier Inc. All rights reserved.

  4. Prognostic Implications of Raphe in Bicuspid Aortic Valve Anatomy.

    Science.gov (United States)

    Kong, William K F; Delgado, Victoria; Poh, Kian Keong; Regeer, Madelien V; Ng, Arnold C T; McCormack, Louise; Yeo, Tiong Cheng; Shanks, Miriam; Parent, Sarah; Enache, Roxana; Popescu, Bogdan A; Liang, Michael; Yip, James W; Ma, Lawrence C W; Kamperidis, Vasileios; van Rosendael, Philippe J; van der Velde, Enno T; Ajmone Marsan, Nina; Bax, Jeroen J

    2017-03-01

    Little is known about the association between bicuspid aortic valve (BAV) morphologic findings and the degree of valvular dysfunction, presence of aortopathy, and complications, including aortic valve surgery, aortic dissection, and all-cause mortality. To investigate the association between BAV morphologic findings (raphe vs nonraphe) and the degree of valve dysfunction, presence of aortopathy, and prognosis (including need for aortic valve surgery, aortic dissection, and all-cause mortality). In this large international multicenter registry of patients with BAV treated at tertiary referral centers, 2118 patients with BAV were evaluated. Patients referred for echocardiography from June 1, 1991, through November 31, 2015, were included in the study. Clinical and echocardiographic data were analyzed retrospectively. The morphologic BAV findings were categorized according to the Sievers and Schmidtke classification. Aortic valve function was divided into normal, regurgitation, or stenosis. Patterns of BAV aortopathy included the following: type 1, dilation of the ascending aorta and aortic root; type 2, isolated dilation of the ascending aorta; and type 3, isolated dilation of the sinus of Valsalva and/or sinotubular junction. Association between the presence and location of raphe and the risk of significant (moderate and severe) aortic valve dysfunction and aortic dilation and/or dissection. Of the 2118 patients (mean [SD] age, 47 [18] years; 1525 [72.0%] male), 1881 (88.8%) had BAV with fusion raphe, whereas 237 (11.2%) had BAV without raphe. Bicuspid aortic valves with raphe had a significantly higher prevalence of valve dysfunction, with a significantly higher frequency of aortic regurgitation (622 [33.1%] vs 57 [24.1%], P < .001) and aortic stenosis (728 [38.7%] vs 51 [21.5%], P < .001). Furthermore, aortic valve replacement event rates were significantly higher among patients with BAV with raphe (364 [19.9%] at 1 year, 393 [21.4%] at 2 years, and 447

  5. Harmane inhibits serotonergic dorsal raphe neurons in the rat.

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    Touiki, Khalid; Rat, Pascal; Molimard, Robert; Chait, Abderrahman; de Beaurepaire, Renaud

    2005-11-01

    Harmane and norharmane (two beta-carbolines) are tobacco components or products. The effects of harmane and norharmane on serotonergic raphe neurons remain unknown. Harmane and norharmane are inhibitors of the monoamine oxidases A (MAO-A) and B (MAO-B), respectively. To study the effects of harmane, norharmane, befloxatone (MAOI-A), and selegiline (MAOI-B) on the firing of serotonergic neurons. To compare the effects of these compounds to those of nicotine (whose inhibitory action on serotonergic neurons has been previously described). The effects of cotinine, a metabolite of nicotine known to interact with serotonergic systems, are also tested. In vivo electrophysiological recordings of serotonergic dorsal raphe neurons in the anaesthetized rat. Nicotine, harmane, and befloxatone inhibited serotonergic dorsal raphe neurons. The other compounds had no effects. The inhibitory effect of harmane (rapid and long-lasting inhibition) differed from that of nicotine (short and rapidly reversed inhibition) and from that of befloxatone (slow, progressive, and long-lasting inhibition). The inhibitory effects of harmane and befloxatone were reversed by the 5-HT1A antagonist WAY 100 635. Pretreatment of animals with p-chlorophenylalanine abolished the inhibitory effect of befloxatone, but not that of harmane. Nicotine, harmane, and befloxatone inhibit the activity of raphe serotonergic neurons. Therefore, at least two tobacco compounds, nicotine and harmane, inhibit the activity of serotonergic neurons. The mechanism by which harmane inhibits serotonergic dorsal raphe neurons is likely unrelated to a MAO-A inhibitory effect.

  6. Development of raphe serotonin neurons from specification to guidance.

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    Kiyasova, Vera; Gaspar, Patricia

    2011-11-01

    The main features of the development of the serotonin (5-HT) raphe neurons have been known for many years but more recent molecular studies, using mouse genetics, have since unveiled several intriguing aspects of the specification of the raphe serotonergic system. These studies indicated that, although all 5-HT neurons in the raphe follow the same general program for their specification, there are also clear regional differences in the way that these neurons are specified and are guided towards different brain targets. Here we overview recent progress made in the understanding of the developmental programming of serotonergic neurons in the mouse raphe, emphasizing data showing how heterogeneous subsets of 5-HT neurons may be generated. Serotonergic progenitors are produced in the brainstem in different rhombomeres under the influence of a set of secreted factors, sonic hedgehog and fibroblast growth factors, which determine their position in the neural tube. Two main transcriptional gene networks are involved in the specification of 5-HT identity, with Lmx1b and Pet1 transcription factors as main players. A differential requirement for Pet1 was, however, revealed, which underlies an anatomical and functional diversity. Transcriptional programs controlling 5-HT identity could also impact axon guidance mechanisms directing 5-HT neurons to their targets. Although no direct links have yet been established, a large set of molecular determinants have already been shown to be involved in the growth, axon guidance and targeting of 5-HT raphe neurons, particularly within the forebrain. Alterations in the molecular mechanisms involved in 5-HT development are likely to have significant roles in mood disease predisposition. © 2011 The Authors. European Journal of Neuroscience © 2011 Federation of European Neuroscience Societies and Blackwell Publishing Ltd.

  7. Geomorphic observations from southwestern terminus of Palghat ...

    Indian Academy of Sciences (India)

    4National Centre for Earth Science Studies, Thiruvananthapuram, Kerala 695 031, India. ∗ .... Geomorphic observations from SW terminus of Palghat Gap, south India. 823. Figure. 2. ... tical point of view to determine an average state.

  8. Dorsal Raphe Dopamine Neurons Represent the Experience of Social Isolation

    Science.gov (United States)

    Matthews, Gillian A.; Nieh, Edward H.; Vander Weele, Caitlin M.; Halbert, Sarah A.; Pradhan, Roma V.; Yosafat, Ariella S.; Glober, Gordon F.; Izadmehr, Ehsan M.; Thomas, Rain E.; Lacy, Gabrielle D.; Wildes, Craig P.; Ungless, Mark A.; Tye, Kay M.

    2016-01-01

    Summary The motivation to seek social contact may arise from either positive or negative emotional states, as social interaction can be rewarding and social isolation can be aversive. While ventral tegmental area (VTA) dopamine (DA) neurons may mediate social reward, a cellular substrate for the negative affective state of loneliness has remained elusive. Here, we identify a functional role for DA neurons in the dorsal raphe nucleus (DRN), in which we observe synaptic changes following acute social isolation. DRN DA neurons show increased activity upon social contact following isolation, revealed by in vivo calcium imaging. Optogenetic activation of DRN DA neurons increases social preference but causes place avoidance. Furthermore, these neurons are necessary for promoting rebound sociability following an acute period of isolation. Finally, the degree to which these neurons modulate behavior is predicted by social rank, together supporting a role for DRN dopamine neurons in mediating a loneliness-like state. PaperClip PMID:26871628

  9. Mesopontine median raphe regulates hippocampal ripple oscillation and memory consolidation.

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    Wang, Dong V; Yau, Hau-Jie; Broker, Carl J; Tsou, Jen-Hui; Bonci, Antonello; Ikemoto, Satoshi

    2015-05-01

    Sharp wave-associated field oscillations (∼200 Hz) of the hippocampus, referred to as ripples, are believed to be important for consolidation of explicit memory. Little is known about how ripples are regulated by other brain regions. We found that the median raphe region (MnR) is important for regulating hippocampal ripple activity and memory consolidation. We performed in vivo simultaneous recording in the MnR and hippocampus of mice and found that, when a group of MnR neurons was active, ripples were absent. Consistently, optogenetic stimulation of MnR neurons suppressed ripple activity and inhibition of these neurons increased ripple activity. Notably, using a fear conditioning procedure, we found that photostimulation of MnR neurons interfered with memory consolidation. Our results demonstrate a critical role of the MnR in regulating ripples and memory consolidation.

  10. Orexin inputs to caudal raphé neurons involved in thermal, cardiovascular, and gastrointestinal regulation.

    Science.gov (United States)

    Berthoud, Hans-Rudolf; Patterson, Laurel M; Sutton, Gregory M; Morrison, Christopher; Zheng, Huiyuan

    2005-02-01

    Orexin-expressing neurons in the lateral hypothalamus with their wide projections throughout the brain are important for the regulation of sleep and wakefulness, ingestive behavior, and the coordination of these behaviors in the environmental context. To further identify downstream effector targets of the orexin system, we examined in detail orexin-A innervation of the caudal raphe nuclei in the medulla, known to harbor sympathetic preganglionic motor neurons involved in thermal, cardiovascular, and gastrointestinal regulation. All three components of the caudal raphe nuclei, raphe pallidus, raphe obscurus, and parapyramidal nucleus, are innervated by orexin-A-immunoreactive fibers. Using confocal microscopy, we demonstrate close anatomical appositions between varicose orexin-A immunoreactive axon profiles and sympathetic premotor neurons identified with either a transneuronal retrograde pseudorabies virus tracer injected into the interscapular brown fat pads, or with in situ hybridization of pro-TRH mRNA. Furthermore, orexin-A injected into the fourth ventricle induced c-Fos expression in the raphe pallidus and parapyramidal nucleus. These findings suggest that orexin neurons in the hypothalamus can modulate brown fat thermogenesis, cardiovascular, and gastrointestinal functions by acting directly on neurons in the caudal raphe nuclei, and support the idea that orexin's simultaneous stimulation of food intake and sympathetic activity might have evolved as a mechanism to stay alert while foraging.

  11. PROJECTIONS OF DORSAL AND MEDIAN RAPHE NUCLEI TO DORSAL AND VENTRAL STRIATUM

    Directory of Open Access Journals (Sweden)

    G. R. Hassanzadeh G. Behzadi

    2007-08-01

    Full Text Available The ascending serotonergic projections are derived mainly from mesencephalic raphe nuclei. Topographical projections from mesencephalic raphe nuclei to the striatum were examined in the rat by the retrograde transport technique of HRP (horseradish peroxidase. In 29 rats stereotaxically injection of HRP enzyme were performed in dorsal and ventral parts of striatum separately. The extent of the injection sites and distribution of retrogradely labeled neuronal cell bodies were drawed on representative sections using a projection microscope. Following ipsilateral injection of HRP into the dorsal striatum, numerous labeled neurons were seen in rostral portion of dorsal raphe (DR nucleus. In the same level the cluster of labeled neurons were hevier through caudal parts of DR. A few neurons were also located in lateral wing of DR. More caudally some labeled neurons were found in lateral, medial line of DR. In median raphe nucleus (MnR the labeled neurons were scattered only in median portion of this nucleus. The ipsilateral injection of HRP into the ventral region of striatum resulted on labeling of numerous neurons in rostral, caudal and lateral portions of DR. Through the caudal extension of DR on 4th ventricle level, a large number of labeled neurons were distributed along the ventrocaudal parts of DR. In MnR, labeled neurons were observed only in median part of this nucleus. These findings suggest the mesencephalic raphe nuclei projections to caudo-putamen are topographically organized. In addition dorsal and median raphe nuclei have a stronger projection to the ventral striatum.

  12. Do dorsal raphe 5-HT neurons encode "beneficialness"?

    Science.gov (United States)

    Luo, Minmin; Li, Yi; Zhong, Weixin

    2016-11-01

    The neurotransmitter serotonin (5-hydroxytryptamine; 5-HT) affects numerous behavioral and physiological processes. Drugs that alter 5-HT signaling treat several major psychiatric disorders and may lead to widespread abuse. The dorsal raphe nucleus (DRN) in the midbrain provides a majority of 5-HT for the forebrain. The importance of 5-HT signaling propels the search for a general theoretical framework under which the diverse functions of the DRN 5-HT neurons can be interpreted and additional therapeutic solutions may be developed. However, experimental data so far support several seeming irreconcilable theories, suggesting that 5-HT neurons mediate behavioral inhibition, aversive processing, or reward signaling. Here, we review recent progresses and propose that DRN 5-HT neurons encode "beneficialness" - how beneficial the current environmental context represents for an individual. Specifically, we speculate that the activity of these neurons reflects the possible net benefit of the current context as determined by p·R-C, in which p indicates reward probability, R the reward value, and C the cost. Through the widespread projections of these neurons to the forebrain, the beneficialness signal may reconfigure neural circuits to bias perception, boost positive emotions, and switch behavioral choices. The "beneficialness" hypothesis can explain many conflicting observations, and at the same time raises new questions. We suggest additional experiments that will help elucidate the exact computational functions of the DRN 5-HT neurons. Copyright © 2016 Elsevier Inc. All rights reserved.

  13. Multiple serotonin receptors: regional distribution and effect of raphe lesions

    International Nuclear Information System (INIS)

    Blackshear, M.A.; Sanders-Bush, E.; Steranka, L.R.

    1981-01-01

    These studies confirm and extend the recent work suggesting that [ 3 H]lysergic acid diethylamide (LSD) labels two distinct binding sites in rat brain resembling serotonin (5HT) receptors. Although Scatchard analyses of [ 3 H]LSD binding to membranes prepared from cortex/hippocampus were linear, the heterogeneity of the [ 3 H]LSD binding sites was clearly demonstrated in displacement studies. The displacement curves for both 5HT and spiperone were bisigmoidal with the concentration required to saturate the high affinity components nearly 3 orders of magnitude lower than the concentrations necessary to saturate the low affinity components. Additivity studies suggested that the sites with high affinity for 5HT and spiperone are different, independent sites. These sites are referred to as 5HT 1 and 5HT 2 respectively. Regional analyses showed, that in the frontal cortex, the density of the 5HT 2 site was slightly greater than the 5HT 1 site whereas the 5HT 1 site was predominant in all other brain areas, including the spinal cord. The pharmacological properties of the two sites have features in common with 5HT receptors; however, electrolytic lesions of the midbrain raphe nuclei did not change the densities or binding constants of the two apparent 5HT receptor subtypes, even though the number of high affinity 5HT uptake sites was markedly reduced. (Auth.)

  14. Dorsal raphe nucleus projecting retinal ganglion cells: Why Y cells?

    Science.gov (United States)

    Pickard, Gary E.; So, Kwok-Fai; Pu, Mingliang

    2015-01-01

    Retinal ganglion Y (alpha) cells are found in retinas ranging from frogs to mice to primates. The highly conserved nature of the large, fast conducting retinal Y cell is a testament to its fundamental task, although precisely what this task is remained ill-defined. The recent discovery that Y-alpha retinal ganglion cells send axon collaterals to the serotonergic dorsal raphe nucleus (DRN) in addition to the lateral geniculate nucleus (LGN), medial interlaminar nucleus (MIN), pretectum and the superior colliculus (SC) has offered new insights into the important survival tasks performed by these cells with highly branched axons. We propose that in addition to its role in visual perception, the Y-alpha retinal ganglion cell provides concurrent signals via axon collaterals to the DRN, the major source of serotonergic afferents to the forebrain, to dramatically inhibit 5-HT activity during orientation or alerting/escape responses, which dis-facilitates ongoing tonic motor activity while dis-inhibiting sensory information processing throughout the visual system. The new data provide a fresh view of these evolutionarily old retinal ganglion cells. PMID:26363667

  15. Ciliated Median Raphe Cyst of Perineum Presenting as Perianal Polyp: A Case Report with Immunohistochemical Study, Review of Literature, and Pathogenesis

    Directory of Open Access Journals (Sweden)

    Jayesh Sagar

    2006-01-01

    Full Text Available Median raphe cyst is a very rare, benign congenital lesion occurring mainly on the ventral aspect of the penis, but can develop anywhere in the midline between the external urethral meatus and anus. We report a case of median raphe cyst in the perineum presenting as a perianal polyp in a 65-year-old, English white male with exceptionally rare ciliated epithelium. According to our knowledge, this is the third such case of ciliated median raphe cyst in the English literature. This case, also the first case of ciliated median raphe cyst in the perineum location, focuses on pathogenesis of median raphe cyst.

  16. Ciliated median raphe cyst of perineum presenting as perianal polyp: a case report with immunohistochemical study, review of literature, and pathogenesis.

    Science.gov (United States)

    Sagar, Jayesh; Sagar, Bethani; Patel, Adam F; Shak, D K

    2006-03-05

    Median raphe cyst is a very rare, benign congenital lesion occurring mainly on the ventral aspect of the penis, but can develop anywhere in the midline between the external urethral meatus and anus. We report a case of median raphe cyst in the perineum presenting as a perianal polyp in a 65-year-old, English white male with exceptionally rare ciliated epithelium. According to our knowledge, this is the third such case of ciliated median raphe cyst in the English literature. This case, also the first case of ciliated median raphe cyst in the perineum location, focuses on pathogenesis of median raphe cyst.

  17. Purification of recombinant C-terminus polyhistidine tagged human ...

    African Journals Online (AJOL)

    Dell

    2012-05-03

    May 3, 2012 ... this research, C-terminus polyhistidine tagged human recombinant calcitonin which was ... range protein molecular weight marker was from SIGMA. PCR- ... supernatant was stored at -80°C until needed for further assays.

  18. Analgesia induced by morphine microinjected into the nucleus raphe magnus: effects on tonic pain.

    Science.gov (United States)

    Dualé, Christian; Sierralta, Fernando; Dallel, Radhouane

    2007-07-01

    One of the possible sites of action of the analgesic effect of morphine is the Nucleus Raphe Magnus, as morphine injected into this structure induces analgesia in transient pain models. In order to test if morphine in the Nucleus Raphe Magnus is also analgesic in a tonic pain model, 5 microg of morphine or saline (control) were microinjected into the Nucleus Raphe Magnus of the rat. Analgesic effects were assessed following nociceptive stimulation using transient heating of the tail (phasic pain) and subcutaneous orofacial injection of 1.5 % formalin (tonic pain). While morphine was strongly analgesic for the tail-flick response (p <0.0001 compared to control), analgesia on the response to formalin was also observed for both early (p = 0.007) and late responses (p = 0.02). However, the response to formalin was not completely blunted. These results suggest that the Nucleus Raphe Magnus is not the exclusive site of action of morphine-induced analgesia in clinical conditions.

  19. Reward Processing by the Dorsal Raphe Nucleus: 5-HT and Beyond

    Science.gov (United States)

    Luo, Minmin; Zhou, Jingfeng; Liu, Zhixiang

    2015-01-01

    The dorsal raphe nucleus (DRN) represents one of the most sensitive reward sites in the brain. However, the exact relationship between DRN neuronal activity and reward signaling has been elusive. In this review, we will summarize anatomical, pharmacological, optogenetics, and electrophysiological studies on the functions and circuit mechanisms of…

  20. Electrophysical properties, synaptic transmission and neuromodulation in serotonergic caudal raphe neurons.

    Science.gov (United States)

    Li, Y W; Bayliss, D A

    1998-06-01

    1. We studied electrophysiological properties, synaptic transmission and modulation by 5-hydroxytryptamine (5-HT) of caudal raphe neurons using whole-cell recording in a neonatal rat brain slice preparation; recorded neurons were identified as serotonergic by post-hoc immunohistochemical detection of tryptophan hydroxylase, the 5-HT-synthesizing enzyme. 2. Serotonergic neurons fired spontaneously (approximately 1 Hz), with maximal steady state firing rates of < 4 Hz. 5-Hydroxytryptamine caused hyperpolarization and cessation of spike activity in these neurons by activating inwardly rectifying K+ conductance via somatodendritic 5-HT1A receptors. 3. Unitary glutamatergic excitatory post-synaptic potentials (EPSP) and currents (EPSC) were evoked in serotonergic neurons by local electrical stimulation. Evoked EPSC were potently inhibited by 5-HT, an effect mediated by presynaptic 5-HT1B receptors. 4. In conclusion, serotonergic caudal raphe neurons are spontaneously active in vitro; they receive prominent glutamatergic synaptic inputs. 5-Hydroxytryptamine regulates serotonergic neuronal activity of the caudal raphe by decreasing spontaneous activity via somatodendritic 5-HT1A receptors and by inhibiting excitatory synaptic transmission onto these neurons via presynaptic 5-HT1B receptors. These local modulatory mechanisms provide multiple levels of feedback autoregulation of serotonergic raphe neurons by 5-HT.

  1. The nucleus raphe interpositus in spinocerebellar ataxia type 3 (Machado-Joseph disease)

    NARCIS (Netherlands)

    Rub, U; Brunt, ER; Gierga, K; Schultz, C; Paulson, H; de Vos, RAI; Braak, H

    The nucleus raphe interpositus (RIP) plays an important role in the premotor network for saccades. Its omnipause neurons gate the activity of the burst neurons for vertical saccades lying within the rostral interstitial nucleus of the medial longitudinal fascicle and that for horizontal saccades

  2. Functional analysis of a novel human serotonin transporter gene promoter in immortalized raphe cells

    DEFF Research Database (Denmark)

    Mortensen, O V; Thomassen, M; Larsen, M B

    1999-01-01

    were found to possess the additional 379 bp fragment. The integrity of the promoter was furthermore confirmed by genomic Southern blotting. The promoter activity was analyzed by reporter gene assays in neuronal and non-neuronal serotonergic cell lines. In immortalized serotonergic raphe neurons, RN46A...

  3. GATA-3 is involved in the development of serotonergic neurons in the caudal raphe nuclei

    NARCIS (Netherlands)

    J. van der Wees (Jacqueline); A. Karis (Alar); E. Goedknegt; M. Rutteman; F.G. Grosveld (Frank); J.H. van Doorninck (Hikke); C.I. de Zeeuw (Chris)

    1999-01-01

    textabstractAbstract The GATA-3 transcription factor shows a specific and restricted expression pattern in the developing and adult mouse brain. In the present study we investigated the role of GATA-3 in the caudal raphe system, which is known to operate as a modulator of motor activity. We

  4. Functional connectivity of the dorsal and median raphe nuclei at rest

    DEFF Research Database (Denmark)

    Beliveau, Vincent; Svarer, Claus; Frokjaer, Vibe G

    2015-01-01

    Serotonin (5-HT) is a neurotransmitter critically involved in a broad range of brain functions and implicated in the pathophysiology of neuropsychiatric illnesses including major depression, anxiety and sleep disorders. Despite being widely distributed throughout the brain, there is limited...... knowledge on the contribution of 5-HT to intrinsic brain activity. The dorsal raphe (DR) and median raphe (MR) nuclei are the source of most serotonergic neurons projecting throughout the brain and thus provide a compelling target for a seed-based probe of resting-state activity related to 5-HT. Here we...... implemented a novel multimodal neuroimaging approach for investigating resting-state functional connectivity (FC) between DR and MR and cortical, subcortical and cerebellar target areas. Using [(11)C]DASB positron emission tomography (PET) images of the brain serotonin transporter (5-HTT) combined...

  5. Estradiol or fluoxetine alters depressive behavior and tryptophan hydroxylase in rat raphe.

    Science.gov (United States)

    Yang, Fu-Zhong; Wu, Yan; Zhang, Wei-Guo; Cai, Yi-Yun; Shi, Shen-Xun

    2010-03-10

    The effects of 17beta-estradiol and fluoxetine on behavior of ovariectomized rats subjected to the forced swimming test and the expression of tryptophan hydroxylase (TPH) in dorsal and median raphe were investigated, respectively through time sampling technique of behavior scoring and immunohistochemistry. Both estradiol and fluoxetine increased swimming and decreased immobility in the forced swimming test. The forced swimming stress decreased integrated optical density of TPH-positive regions in dorsal and median raphe. Both estradiol and fluoxetine administration prevented integrated optical density of TPH-positive regions from being decreased by forced swimming stress. These observations suggest that both estradiol and fluoxetine have protective bearing on ovariectomized rats enduring forced swimming stress.

  6. [Local GABA-ergic modulation of serotonergic neuron activity in the nucleus raphe magnus].

    Science.gov (United States)

    Iniushkin, A N; Merkulova, N A; Orlova, A O; Iniushkina, E M

    2009-07-01

    In voltage-clamp experimental on slices of the rat brainstem the effects of 5-HT and GABA on serotonergic neurons of nucleus raphe magnus were investigated. Local applications of 5-HT induced an increase in IPCSs frequency and amplitude in 45% of serotonergic cells. The effect suppressed by the blocker of fast sodium channels tetradotoxin. Antagonist of GABA receptor gabazine blocked IPSCs in neurons both sensitive and non-sensitive to 5-HT action. Applications of GABA induced a membrane current (I(GABA)), which was completely blocked by gabazine. The data suggest self-control of the activity of serotonergic neurons in nucleus raphe magnus by negative feedback loop via local GABAergic interneurons.

  7. Extrasynaptic glycine receptors of rodent dorsal raphe serotonergic neurons:a sensitive target for ethanol

    OpenAIRE

    Maguire, Edward P.; Mitchell, Elizabeth A.; Greig, Scott J.; Corteen, Nicole; Balfour, David J. K.; Swinny, Jerome; Lambert, Jeremy J.; Belelli, Delia

    2014-01-01

    Alcohol abuse is a significant medical and social problem. Several neurotransmitter systems are implicated in ethanol's actions, with certain receptors and ion channels emerging as putative targets. The dorsal raphe (DR) nucleus is associated with the behavioral actions of alcohol, but ethanol actions on these neurons are not well understood. Here, using immunohistochemistry and electrophysiology we characterize DR inhibitory transmission and its sensitivity to ethanol. DR neurons exhibit inh...

  8. Creating standards: Creating illusions?

    DEFF Research Database (Denmark)

    Linneberg, Mai Skjøtt

    written standards may open up for the creation of illusions. These are created when written standards' content is not in accordance with the perception standard adopters and standard users have of the specific practice phenomenon's content. This general theoretical argument is exemplified by the specific...

  9. Hpa1 harpin needs nitroxyl terminus to promote vegetative growth ...

    Indian Academy of Sciences (India)

    2016-08-26

    Aug 26, 2016 ... Home; Journals; Journal of Biosciences; Volume 39; Issue 1. Hpa1 harpin needs nitroxyl terminus to ... Hansong Dong1 Chunling Zhang1. State Ministry of Education Key Laboratory of Integrated Management of Crop Pathogens and Insect Pests, Nanjing Agricultural University, Jiangsu, 210095, China ...

  10. Single-prolonged stress induces apoptosis in dorsal raphe nucleus in the rat model of posttraumatic stress disorder

    Directory of Open Access Journals (Sweden)

    Liu Dongjuan

    2012-11-01

    Full Text Available Abstract Introduction Post-traumatic stress disorder (PTSD is an anxiety disorder that develops after exposure to a life-threatening traumatic experience. Meta-analyses of the brainstem showed that midsagittal area of the pons was significantly reduced in patients with PTSD, suggesting a potential apoptosis in dorsal raphe nucleus after single-prolonged stress (SPS. The aim of this study is to investigate whether SPS induces apoptosis in dorsal raphe nucleus in PTSD rats, which may be a possible mechanism of reduced volume of pons and density of gray matter. Methods In this study, rats were randomly divided into 1d, 7d and 14d groups after SPS along with the control group. The apoptosis rate was determined using annexin V-FITC/PI double-labeled flow cytometry (FCM. Levels of Cytochrome c (Cyt-C was examined by Western blotting. Expression of Cyt-C on mitochondria in the dorsal raphe nucleus neuron was determined by enzymohistochemistry under transmission electron microscopy (TEM. The change of thiamine monophosphatase (TMP levels was assessed by enzymohistochemistry under light microscope and TEM. Morphological changes of the ultrastructure of the dorsal raphe nucleus neuron were determined by TEM. Results Apoptotic morphological alterations were observed in dorsal raphe nucleus neuron for all SPS-stimulate groups of rats. The apoptosis rates were significantly increased in dorsal raphe nucleus neuron of SPS rats, along with increased release of cytochrome c from the mitochondria into the cytoplasm, increased expression of Cyt-C and TMP levels in the cytoplasm, which reached to the peak of increase 7 days of SPS. Conclusions The results indicate that SPS induced Cyt-C released from mitochondria into cytosol and apoptosis in dorsal raphe nucleus neuron of rats. Increased TMP in cytoplasm facilitated the clearance of apoptotic cells. We propose that this presents one of the mechanisms that lead to reduced volume of pons and gray matter associated

  11. Nitric oxide in the nucleus raphe magnus modulates cutaneous blood flow in rats during hypothermia

    Directory of Open Access Journals (Sweden)

    Masoumeh Kourosh Arami

    2015-10-01

    Full Text Available Objective(s: Nucleus Raphe Magnus (NRM that is involved in the regulation of body temperature contains nitric oxide (NO synthase. Considering the effect of NO on skin blood flow control, in this study, we assessed its thermoregulatory role within the raphe magnus. Materials and Methods: To this end, tail blood flow of male Wistar rats was measured by laser doppler following the induction of hypothermia. Results: Intra-NRM injection of SNP (exogenous NO donor, 0.1- 0.2 μl, 0.2 nM increased the blood flow. Similarly, unilateral microinjection of glutamate (0.1- 0.2 μl, 2.3 nM into the nucleus increased the blood flow. This effectof L-glutamate was reduced by prior intra NRM administrationof NO synthase inhibitor NG-methyl-L-arginine or NG-nitro-L-argininemethyl ester (L-NAME, 0.1 µl, 100 nM. Conclusion: It is concluded that NO modulates the thermoregulatory response of NRM to hypothermia and may interactwith excitatory amino acids in central skin blood flow regulation.

  12. Nitric oxide in the nucleus raphe magnus modulates cutaneous blood flow in rats during hypothermia.

    Science.gov (United States)

    Arami, Masoumeh Kourosh; Zade, Javad Mirnajafi; Komaki, Alireza; Amiri, Mahmood; Mehrpooya, Sara; Jahanshahi, Ali; Jamei, Behnam

    2015-10-01

    Nucleus Raphe Magnus (NRM) that is involved in the regulation of body temperature contains nitric oxide (NO) synthase. Considering the effect of NO on skin blood flow control, in this study, we assessed its thermoregulatory role within the raphe magnus. To this end, tail blood flow of male Wistar rats was measured by laser doppler following the induction of hypothermia. Intra-NRM injection of SNP (exogenous NO donor, 0.1- 0.2 μl, 0.2 nM) increased the blood flow. Similarly, unilateral microinjection of glutamate (0.1- 0.2 μl, 2.3 nM) into the nucleus increased the blood flow. This effect of L-glutamate was reduced by prior intra NRM administration of NO synthase inhibitor N(G)-methyl-L-arginine or N(G)-nitro-L-arginine methyl ester (L-NAME, 0.1 µl, 100 nM). It is concluded that NO modulates the thermoregulatory response of NRM to hypothermia and may interact with excitatory amino acids in central skin blood flow regulation.

  13. Role of the synaptobrevin C terminus in fusion pore formation

    DEFF Research Database (Denmark)

    Ngatchou, Annita N; Kisler, Kassandra; Fang, Qinghua

    2010-01-01

    Neurotransmitter release is mediated by the SNARE proteins synaptobrevin II (sybII, also known as VAMP2), syntaxin, and SNAP-25, generating a force transfer to the membranes and inducing fusion pore formation. However, the molecular mechanism by which this force leads to opening of a fusion pore...... stimulation, the SNARE complex pulls the C terminus of sybII deeper into the vesicle membrane. We propose that this movement disrupts the vesicular membrane continuity leading to fusion pore formation. In contrast to current models, the experiments suggest that fusion pore formation begins with molecular...

  14. [Influence of estradiol on tryptophan hydroxylase and 5-hydroxytryptamine content in raphe nuclei of rats under forced swimming stress].

    Science.gov (United States)

    Yang, Fu-zhong; Wu, Yan; Zhang, Wei-guo; Cai, Yi-yun; Shi, Shen-xun

    2010-07-20

    To investigate the effect of estradiol (E2) on tryptophan hydroxylase (TPH) and 5-hydroxytryptamine (5-HT) content in raphe nuclei of rats under forced swimming stress and explore the role of estrogen and stress in disease mechanism of depression in women. At Week 3 post-ovariectomy, 35 ovariectomized (OVX) female SD rats were randomly divided into 5 groups (n = 7): non-stress group, control group, estradiol (E2) group and fluoxetine (FLX) group and E2 plus FLX group. Animals were administered with different drugs for 2 weeks. At Day 14, animals except those in the non-stress group were subjected to the 15 min forced swimming test (FST). At 2 hours post-FST, all animals including those in the non-stress group were perfused with 4% paraformaldehyde and brains removed for TPH and 5-HT immunofluorescence staining. We compared the content of TPH and 5-HT by observing and calculating the integrated optical density (IOD) of immunofluorescent-positive signals in raphe nuclei. (1) The IOD value of TPH- and 5-HT-positive region in raphe nuclei of rats in the control group was significantly lower than that of the non-stress group (P Forced swimming stress can decrease the TPH and 5-HT content in raphe nuclei. Such changes can be prevented by a pre-administration of estradiol. Similar results are observed with antidepressant fluoxetine. These effects may underlie the role of estradiol and stress in the disease mechanism of depression in women.

  15. Serotonergic projections from the raphe nuclei to the subthalamic nucleus; a retrograde- and anterograde neuronal tracing study

    DEFF Research Database (Denmark)

    Reznitsky, Martin; Plenge, Per; Hay-Schmidt, Anders

    2016-01-01

    the 5-HT1A and 5-HT2A not were present. Retrograde tracer FluoroGold or Choleratoxin subunit B were iontophoretically delivered in the STN and combined with immunohistochemistry for 5-HT in order to map the topographic organization in the dorsal raphe system. The study showed that approximately 320...

  16. Raphe serotonin neuron-specific oxytocin receptor knockout reduces aggression without affecting anxiety-like behavior in male mice only.

    Science.gov (United States)

    Pagani, J H; Williams Avram, S K; Cui, Z; Song, J; Mezey, É; Senerth, J M; Baumann, M H; Young, W S

    2015-02-01

    Serotonin and oxytocin influence aggressive and anxiety-like behaviors, though it is unclear how the two may interact. That the oxytocin receptor is expressed in the serotonergic raphe nuclei suggests a mechanism by which the two neurotransmitters may cooperatively influence behavior. We hypothesized that oxytocin acts on raphe neurons to influence serotonergically mediated anxiety-like, aggressive and parental care behaviors. We eliminated expression of the oxytocin receptor in raphe neurons by crossing mice expressing Cre recombinase under control of the serotonin transporter promoter (Slc6a4) with our conditional oxytocin receptor knockout line. The knockout mice generated by this cross are normal across a range of behavioral measures: there are no effects for either sex on locomotion in an open-field, olfactory habituation/dishabituation or, surprisingly, anxiety-like behaviors in the elevated O and plus mazes. There was a profound deficit in male aggression: only one of 11 raphe oxytocin receptor knockouts showed any aggressive behavior, compared to 8 of 11 wildtypes. In contrast, female knockouts displayed no deficits in maternal behavior or aggression. Our results show that oxytocin, via its effects on raphe neurons, is a key regulator of resident-intruder aggression in males but not maternal aggression. Furthermore, this reduction in male aggression is quite different from the effects reported previously after forebrain or total elimination of oxytocin receptors. Finally, we conclude that when constitutively eliminated, oxytocin receptors expressed by serotonin cells do not contribute to baseline anxiety-like behaviors or maternal care. © 2015 John Wiley & Sons Ltd and International Behavioural and Neural Genetics Society.

  17. Disconnectivity between Dorsal Raphe Nucleus and Posterior Cingulate Cortex in Later Life Depression

    Directory of Open Access Journals (Sweden)

    Toshikazu Ikuta

    2017-08-01

    Full Text Available The dorsal raphe nucleus (DRN has been repeatedly implicated as having a significant relationship with depression, along with its serotoninergic innervation. However, functional connectivity of the DRN in depression is not well understood. The current study aimed to isolate functional connectivity of the DRN distinct in later life depression (LLD compared to a healthy age-matched population. Resting state functional magnetic resonance imaging (rsfMRI data from 95 participants (33 LLD and 62 healthy were collected to examine functional connectivity from the DRN to the whole brain in voxel-wise fashion. The posterior cingulate cortex (PCC bilaterally showed significantly smaller connectivity in the LLD group than the control group. The DRN to PCC connectivity did not show any association with the depressive status. The findings implicate that the LLD involves disruption of serotoninergic input to the PCC, which has been suggested to be a part of the reduced default mode network in depression.

  18. Effect of arginine vasopressin in the nucleus raphe magnus on antinociception in the rat.

    Science.gov (United States)

    Yang, Jun; Chen, Jian-Min; Liu, Wen-Yan; Song, Cao-You; Wang, Cheng-Hai; Lin, Bao-Cheng

    2006-09-01

    Previous work has shown that arginine vasopressin (AVP) regulates antinociception through brain nuclei rather than the spinal cord and peripheral organs. The present study investigated the nociceptive effect of AVP in the nucleus raphe magnus (NRM) of the rat. Microinjection of AVP into the NRM increased pain threshold in a dose-dependent manner, while local administration of AVP-receptor antagonist-d(CH2)5Tyr(Et)DAVP decreased the pain threshold. Pain stimulation elevated AVP concentration in the NRM perfuse liquid. NRM pretreatment with AVP-receptor antagonist completely reversed AVP's effect on pain threshold in the NRM. The data suggest that AVP in the NRM is involved in antinociception.

  19. Construction and expression of eukaryotic expression vectors of full-length, amino-terminus and carboxyl-terminus Raf gene

    Directory of Open Access Journals (Sweden)

    Zhuomin WANG

    2008-06-01

    Full Text Available Background and objective Raf is a key molecule in the Ras-Raf-MEK-ERK signal transduction pathway and is highly activated in different human carcinomas. However, its biological functions and regulation mechanisms are still unclear. The aims of this study were to construct eukaryotic expression vectors with Raf full encoding region, truncated amino-terminus and carboxyl-terminus, respectively. Methods Eukaryotic expression vectors of pCMV-Tag2b-Raf-1, pCMV-Tag2b-N-Raf and pCMV-Tag2b-C-Raf were constructed by gene recombination technique and confirmed by restriction enzyme analysis and DNA sequencing. Furthermore, the expression of these fusion proteins was detected by western blot in transient transfected 293T cells. Results The sequences and open reading frames of these three vectors were completely consistent with experimental design. All target proteins can be detected in 293T cells. Conclusion Eukaryotic expression vectors of pCMV-Tag2b-Raf-1, pCMV-Tag2b-N-Raf and pCMV-Tag2b-C-Raf were successfully constructed and can be expressed in 293T cells.

  20. An in-vitro comparison of the radiographic and actual gutta-percha terminus.

    Science.gov (United States)

    Namazikhah, M S; Ghiai, M; Parkin, M J; Puccinelli, L

    2000-06-01

    The purpose of this study was to investigate the difference between the radiographic gutta-percha terminus and the actual gutta-percha terminus of human molars by comparing radiographic obturation results with actual obturation results. Forty maxillary palatal roots and 50 mandibular distal roots were randomly selected from a population of 540. They were then mounted in stone and radiographed. Conventional endodontic therapy was completed using stainless-steel K files and lateral condensation. Each radiographic gutta-percha terminus was evaluated under 4.5x magnification by three examiners following the completion of root canal therapy. These results were recorded. Each tooth was then removed from its mounting, and the actual gutta-percha terminus was evaluated under 4.5x magnification. These results were recorded and compared to the radiographic gutta-percha terminus results. In all 90 teeth examined, the actual gutta-percha terminus was equal to or longer than the radiographic gutta-percha terminus. In the 50 mandibular distal roots, the actual gutta-percha terminus averaged 0.645 mm longer than the radiographic gutta-percha terminus. In the 40 maxillary palatal roots, this difference measured 0.6375 mm.

  1. Self-transcendence trait and its relationship with in vivo serotonin transporter availability in brainstem raphe nuclei: An ultra-high resolution PET-MRI study.

    Science.gov (United States)

    Kim, Jong-Hoon; Son, Young-Don; Kim, Jeong-Hee; Choi, Eun-Jung; Lee, Sang-Yoon; Joo, Yo-Han; Kim, Young-Bo; Cho, Zang-Hee

    2015-12-10

    Self-transcendence is an inherent human personality trait relating to the experience of spiritual aspects of the self. We examined the relationship between self-transcendence and serotonin transporter (SERT) availability in brainstem raphe nuclei, which are collections of five different serotonergic nuclei with rostro-caudal extension, using ultra-high resolution magnetic resonance imaging (MRI) and positron emission tomography (PET) with (11)C-3-amino-4-(2-dimethylaminomethylphenylthio)benzonitrile ([(11)C]DASB) to elucidate potential roles of serotonergic neuronal activities in this personality trait. Sixteen healthy subjects completed 7.0T MRI and High Resolution Research Tomograph (HRRT) PET. The regions of interest (ROIs) included the dorsal raphe nucleus (R1), median raphe nucleus (R2), raphe pontis (R3), and the caudal raphe nuclei (R4 and R5). For the estimation of SERT availability, the binding potential (BPND) was derived using the simplified reference tissue model (SRTM2). The Temperament and Character Inventory was used to measure self-transcendence. The analysis revealed that the self-transcendence total score had a significant negative correlation with the [(11)C]DASB BPND in the caudal raphe (R5). The subscale score for spiritual acceptance was significantly negatively correlated with the [(11)C]DASB BPND in the median raphe nucleus (R2). The results indicate that the self-transcendence trait is associated with SERT availability in specific raphe subnuclei, suggesting that the serotonin system may serve as an important biological basis for human self-transcendence. Based on the connections of these nuclei with cortico-limbic and visceral autonomic structures, the functional activity of these nuclei and their related neural circuitry may play a crucial role in the manifestation of self-transcendence. Copyright © 2015. Published by Elsevier B.V.

  2. Fos and serotonin immunoreactivity in the raphe nuclei of the cat during carbachol-induced active sleep: a double-labeling study.

    Science.gov (United States)

    Yamuy, J; Sampogna, S; López-Rodríguez, F; Luppi, P H; Morales, F R; Chase, M H

    1995-07-01

    The microinjection of carbachol into the nucleus pontis oralis produces a state which is polygraphically and behaviorally similar to active sleep (rapid eye movement sleep). In the present study, using double-labeling techniques for serotonin and the protein product of c-fos (Fos), we sought to examine whether immunocytochemically identified serotonergic neurons of the raphe nuclei of the cat were activated, as indicated by their expression of c-fos, during this pharmacologically-induced behavioral state (active sleep-carbachol). Compared with control cats, which were injected with saline, active sleep-carbachol cats exhibited a significantly greater number of c-fos-expressing neurons in the raphe dorsalis, magnus and pallidus. Whereas most of the c-fos-expressing neurons in the raphe dorsalis were small, those in the raphe magnus were medium-sized and in the raphe pallidus they were small and medium-sized. The mean number of serotonergic neurons that expressed c-fos (i.e. double-labeled cells) was similar in control and active sleep-carbachol cats. These data indicate that there is an increased number of non-serotonergic, c-fos-expressing neurons in the raphe dorsalis, magnus and pallidus during the carbachol-induced state.(ABSTRACT TRUNCATED AT 250 WORDS)

  3. Detecting frontal ablation processes from direct observations of submarine terminus morphology

    Science.gov (United States)

    Fried, M.; Carroll, D.; Catania, G. A.; Sutherland, D. A.; Stearns, L. A.; Bartholomaus, T. C.; Shroyer, E.; Nash, J. D.

    2017-12-01

    Tidewater glacier termini couple glacier and ocean systems. Subglacial discharge emerging from the terminus produces buoyant plumes that modulate submarine melting, calving, fjord circulation and, in turn, changes in ice dynamics from back-stress perturbations. However, the absence of critical observational data at the ice-ocean interface limits plume and, by extension, melt models from incorporating realistic submarine terminus face morphologies and assessing their impact on terminus behavior at tidewater glaciers. Here we present a comprehensive inventory and characterization of submarine terminus face shapes from a side-looking, multibeam echo sounding campaign across Kangerdlugssuaq Sermerssua glacier, central-west Greenland. We combine these observations with in-situ measurements of ocean stratification and remotely sensed subglacial discharge, terminus positions, ice velocity, and ice surface datasets to infer the spectrum of processes sculpting the submarine terminus face. Subglacial discharge outlet locations are confirmed through observations of sediment plumes, localized melt-driven undercutting of the terminus face, and bathymetry of the adjacent seafloor. From our analysis, we differentiate terminus morphologies resulting from submarine melt and calving and assess the contribution of each process to the net frontal ablation budget. Finally, we constrain a plume model using direct observations of the submarine terminus face and conduit geometry. Plume model simulations demonstrate that the majority of discharge outlets are fed by small discharge fluxes, suggestive of a distributed subglacial hydrologic system. Outlets with the largest, concentrated discharge fluxes are morphologically unique and strongly control seasonal terminus position. At these locations, we show that the spatiotemporal pattern of terminus retreat is well correlated with time periods when local melt rate exceeds ice velocity.

  4. C-Terminal carbohydrate-binding module 9_2 fused to the N-terminus of GH11 xylanase from Aspergillus niger.

    Science.gov (United States)

    Xu, Wenxuan; Liu, Yajuan; Ye, Yanxin; Liu, Meng; Han, Laichuang; Song, Andong; Liu, Liangwei

    2016-10-01

    The 9_2 carbohydrate-binding module (C2) locates natively at the C-terminus of the GH10 thermophilic xylanase from Thermotoga marimita. When fused to the C-terminus, C2 improved thermostability of a GH11 xylanase (Xyn) from Aspergillus niger. However, a question is whether the C-terminal C2 would have a thermostabilizing effect when fused to the N-terminus of a catalytic module. A chimeric enzyme, C2-Xyn, was created by step-extension PCR, cloned in pET21a(+), and expressed in E. coli BL21(DE3). The C2-Xyn exhibited a 2 °C higher optimal temperature, a 2.8-fold longer thermostability, and a 4.5-fold higher catalytic efficiency on beechwood xylan than the Xyn. The C2-Xyn exhibited a similar affinity for binding to beechwood xylan and a higher affinity for oat-spelt xylan than Xyn. C2 is a thermostabilizing carbohydrate-binding module and provides a model of fusion at an enzymatic terminus inconsistent with the modular natural terminal location.

  5. Effects of obesogenic diet and estradiol on dorsal raphe gene expression in old female macaques.

    Directory of Open Access Journals (Sweden)

    Cynthia L Bethea

    -induced gene expression in the dorsal raphe. These results suggest that with regard to function and viability in the dorsal raphe, HT may not be as beneficial for obese women as normal weight women.

  6. Increased mRNA expression of cytochrome oxidase in dorsal raphe nucleus of depressive suicide victims

    Directory of Open Access Journals (Sweden)

    A Sanchez-Bahillo

    2008-04-01

    Full Text Available A Sanchez-Bahillo1, V Bautista-Hernandez1, Carlos Barcia Gonzalez1, R Bañon2, A Luna2, EC Hirsch3, Maria-Trinidad Herrero11Clinical and Experimental Neuroscience, Centro de Investigación Biomédica en Red sobre Enfermedades Neurodegenerativas (CIBERNED; 2Department of Legal Medicine, Department of Human Anatomy, School of Medicine, University of Murcia, Campus de Espinardo, Murcia 30100, Spain; 3INSERM U679 Hôpital de la Salpêtrière, Boulevard de l’Hôpital, Paris, FranceAbstract: Suicidal behavior is a problem with important social repercussions. Some groups of the population show a higher risk of suicide; for example, depression, alcoholism, psychosis or drug abuse frequently precedes suicidal behavior. However, the relationship between metabolic alterations in the brain and premorbid clinical symptoms of suicide remains uncertain. The serotonergic and noradrenergic systems have frequently been, implicated in suicidal behavior and the amount of serotonin in the brain and CSF of suicide victims has been found to be low compared with normal subjects. However, there are contradictory results regarding the role of noradrenergic neurons in the mediation of suicide attempts, possibly reflecting the heterogeneity of conditions that lead to a common outcome. In the present work we focus on the subgroup of suicide victims that share a common diagnosis of major depression. Based on post-mortem studies analyzing mRNA expression by in situ hybridization, serotonergic neurons from the dorsal raphe nucleus (DRN from depressive suicide victims are seen to over-express cytochrome oxidase mRNA. However, no corresponding changes were found in the expression of tyrosine hydroxylase (TH mRNA in the noradrenergic neurons of the Locus Coeruleus (LC. These results suggest that, despite of the low levels of serotonin described in suicide victims, the activity of DRN neurons could increase in the suicidally depressed, probably due to the over activation of

  7. Distinct Contributions of Median Raphe Nucleus to Contextual Fear Conditioning and Fear-Potentiated Startle

    Science.gov (United States)

    Silva, R. C. B.; Cruz, A. P. M.; Avanzi, V.; Landeira-Fernandez, J.; Brandão, M. L.

    2002-01-01

    Ascending 5-HT projections from the median raphe nucleus (MRN), probably to the hippocampus, are implicated in the acquisition of contextual fear (background stimuli), as assessed by freezing behavior. Foreground cues like light, used as a conditioned stimulus (CS) in classical fear conditioning, also cause freezing through thalamic transmission to the amygdala. As the MRN projects to the hippocampus and amygdala, the role of this raphe nucleus in fear conditioning to explicit cues remains to be explained. Here we analyzed the behavior of rats with MRN electrolytic lesions in a contextual conditioning situation and in a fear-potentiated startle procedure. The animals received MRN electrolytic lesions either before or on the day after two consecutive training sessions in which they were submitted to 10 conditioning trials, each in an experimental chamber (same context) where they. received foot-shocks (0.6 mA, 1 sec) paired to a 4-sec light CS. Seven to ten days later, the animals were submitted to testing sessions for assessing conditioned fear when they were placed for five shocks, and the duration of contextual freezing was recorded. The animals were then submitted to a fear-potentiated startle in response to a 4-sec light-CS, followed by white noise (100 dB, 50 ms). Control rats (sham) tested in the same context showed more freezing than did rats with pre- or post-training MRN lesions. Startle was clearly potentiated in the presence of light CS in the sham-lesioned animals. Whereas pretraining lesions reduced both freezing and fear-potentiated startle, the post-training lesions reduced only freezing to context, without changing the fear-potentiated startle. In a second experiment, neurotoxic lesions of the MRN with local injections of N-methyl-D-aspartate or the activation of 5-HT1A somatodendritic auto-receptors of the MRN by microinjections of the 5-HT1A receptor agonist 8-hydroxy- 2-(di-n-propylamino)tetralin (8-OH-DPAT) before the training sessions also

  8. Positive regulation of raphe serotonin neurons by serotonin 2B receptors.

    Science.gov (United States)

    Belmer, Arnauld; Quentin, Emily; Diaz, Silvina L; Guiard, Bruno P; Fernandez, Sebastian P; Doly, Stéphane; Banas, Sophie M; Pitychoutis, Pothitos M; Moutkine, Imane; Muzerelle, Aude; Tchenio, Anna; Roumier, Anne; Mameli, Manuel; Maroteaux, Luc

    2018-06-01

    Serotonin is a neurotransmitter involved in many psychiatric diseases. In humans, a lack of 5-HT 2B receptors is associated with serotonin-dependent phenotypes, including impulsivity and suicidality. A lack of 5-HT 2B receptors in mice eliminates the effects of molecules that directly target serotonergic neurons including amphetamine derivative serotonin releasers, and selective serotonin reuptake inhibitor antidepressants. In this work, we tested the hypothesis that 5-HT 2B receptors directly and positively regulate raphe serotonin neuron activity. By ex vivo electrophysiological recordings, we report that stimulation by the 5-HT 2B receptor agonist, BW723C86, increased the firing frequency of serotonin Pet1-positive neurons. Viral overexpression of 5-HT 2B receptors in these neurons increased their excitability. Furthermore, in vivo 5-HT 2B -receptor stimulation by BW723C86 counteracted 5-HT 1A autoreceptor-dependent reduction in firing rate and hypothermic response in wild-type mice. By a conditional genetic ablation that eliminates 5-HT 2B receptor expression specifically and exclusively from Pet1-positive serotonin neurons (Htr2b 5-HTKO mice), we demonstrated that behavioral and sensitizing effects of MDMA (3,4-methylenedioxy-methamphetamine), as well as acute behavioral and chronic neurogenic effects of the antidepressant fluoxetine, require 5-HT 2B receptor expression in serotonergic neurons. In Htr2b 5-HTKO mice, dorsal raphe serotonin neurons displayed a lower firing frequency compared to control Htr2b lox/lox mice as assessed by in vivo extracellular recordings and a stronger hypothermic effect of 5-HT 1A -autoreceptor stimulation was observed. The increase in head-twitch response to DOI (2,5-dimethoxy-4-iodoamphetamine) further confirmed the lower serotonergic tone resulting from the absence of 5-HT 2B receptors in serotonin neurons. Together, these observations indicate that the 5-HT 2B receptor acts as a direct positive modulator of serotonin Pet1

  9. Confirmation of a blocked amino terminus of sulfhydryl oxidase

    International Nuclear Information System (INIS)

    Janolino, V.G.; Morrison-Rowe, S.J.; Swaisgood, H.E.

    1990-01-01

    The isolation of sulfhydryl oxidase from bovine milk in a suitably pure form for sequencing was carried out by transient covalent affinity chromatography of diafiltered whey using cysteinylsuccinamidopropyl-glass as matrix. The glutathione-eluted proteins were separated by SDS-PAGE. By radiolabeling the affinity chromatography-purified enzyme with [ 14 C]iodoacetate before subjecting to SDS-PAGE, the sulfhydryl oxidase band was identified, because sulfhydryl oxidase is known to be inactivated by alkylation of one sulfhydryl group per mole. The results confirmed that sulfhydryl oxidase corresponds to the 85 (± 5)-kDa band observed on SDS-PAGE. The protein band corresponding to radiolabeled sulfhydryl oxidase was recovered from SDS-PAGE gels by electrophoretic elution and by electroblotting on polyvinylidene difluoride membrane and subjected to gas phase sequencing. Precautions were taken during electrophoretic elution to prevent reactions that result in N-terminal blocking. Both methods of protein recovery yielded negative results when subjected to sequence analysis indicating that the N-terminus of sulfhydryl oxidase is blocked

  10. Serotonergic versus Nonserotonergic Dorsal Raphe Projection Neurons: Differential Participation in Reward Circuitry

    Directory of Open Access Journals (Sweden)

    Ross A. McDevitt

    2014-09-01

    Full Text Available The dorsal raphe nucleus (DRN contains the largest group of serotonin-producing neurons in the brain and projects to regions controlling reward. Although pharmacological studies suggest that serotonin inhibits reward seeking, electrical stimulation of the DRN strongly reinforces instrumental behavior. Here, we provide a targeted assessment of the behavioral, anatomical, and electrophysiological contributions of serotonergic and nonserotonergic DRN neurons to reward processes. To explore DRN heterogeneity, we used a simultaneous two-vector knockout/optogenetic stimulation strategy, as well as cre-induced and cre-silenced vectors in several cre-expressing transgenic mouse lines. We found that the DRN is capable of reinforcing behavior primarily via nonserotonergic neurons, for which the main projection target is the ventral tegmental area (VTA. Furthermore, these nonserotonergic projections provide glutamatergic excitation of VTA dopamine neurons and account for a large majority of the DRN-VTA pathway. These findings help to resolve apparent discrepancies between the roles of serotonin versus the DRN in behavioral reinforcement.

  11. Biophysical properties and computational modeling of calcium spikes in serotonergic neurons of the dorsal raphe nucleus.

    Science.gov (United States)

    Tuckwell, Henry C

    2013-06-01

    Serotonergic neurons of the dorsal raphe nuclei, with their extensive innervation of nearly the whole brain have important modulatory effects on many cognitive and physiological processes. They play important roles in clinical depression and other psychiatric disorders. In order to quantify the effects of serotonergic transmission on target cells it is desirable to construct computational models and to this end these it is necessary to have details of the biophysical and spike properties of the serotonergic neurons. Here several basic properties are reviewed with data from several studies since the 1960s to the present. The quantities included are input resistance, resting membrane potential, membrane time constant, firing rate, spike duration, spike and afterhyperpolarization (AHP) amplitude, spike threshold, cell capacitance, soma and somadendritic areas. The action potentials of these cells are normally triggered by a combination of sodium and calcium currents which may result in autonomous pacemaker activity. We here analyse the mechanisms of high-threshold calcium spikes which have been demonstrated in these cells the presence of TTX (tetrodotoxin). The parameters for calcium dynamics required to give calcium spikes are quite different from those for regular spiking which suggests the involvement of restricted parts of the soma-dendritic surface as has been found, for example, in hippocampal neurons. Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.

  12. Tolerance to non-opioid analgesics is opioid-sensitive in nucleus raphe magnus

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    Merab G Tsagareli

    2011-07-01

    Full Text Available Repeated injection of opioid analgesics can lead to a progressive loss of its effect. This phenomenon is known as tolerance. Several lines of investigations have shown that systemic, intraperitoneal administration or the microinjection of non-opioid analgesics, non-steroidal anti-inflammatory drugs (NSAIDs in the midbrain periaqueductal gray matter induces antinociception with some effects of tolerance. Our recent study has revealed that microinjection of three drugs analgin, ketorolac and xefocam into the central nucleus of amygdala produce tolerance to them and cross-tolerance to morphine. Here we report that repeated administrations of these NSAIDs into the nucleus raphe magnus (NRM in the following four days result in progressively less antinociception, i.e. produce the development of tolerance to these drugs in mail rats. Special control experiments showed that post-treatment with μ-opioid antagonist naloxone in NRM significantly decreased antinociceptive effects of NSAIDs at the first day in behavioral tail flick reflex (TF and hot plate (HP latencies. At the second day, naloxone generally had trend effects in both TF and HP tests impeded the development of tolerance to the antinociceptive effect of non-opioid analgesics. These findings strongly support the suggestion on endogenous opioid involvement in NSAIDs antinociception and tolerance in the descending pain control system. Moreover, repeated injections of NSAIDs progressively lead to tolerance to them, cross-tolerance to morphine and the risk of a withdrawal syndrome. Therefore, these results are important for human medicine too.

  13. Opioid microinjection into raphe magnus modulates cardiorespiratory function in mice and rats.

    Science.gov (United States)

    Hellman, Kevin M; Mendelson, Scott J; Mendez-Duarte, Marco A; Russell, James L; Mason, Peggy

    2009-11-01

    The raphe magnus (RM) participates in opioid analgesia and contains pain-modulatory neurons with respiration-related discharge. Here, we asked whether RM contributes to respiratory depression, the most prevalent lethal effect of opioids. To investigate whether opioidergic transmission in RM produces respiratory depression, we microinjected a mu-opioid receptor agonist, DAMGO, or morphine into the RM of awake rodents. In mice, opioid microinjection produced sustained decreases in respiratory rate (170 to 120 breaths/min), as well as heart rate (520 to 400 beats/min). Respiratory sinus arrhythmia, indicative of enhanced parasympathetic activity, was prevalent in mice receiving DAMGO microinjection. We performed similar experiments in rats but observed no changes in breathing rate or heart rate. Both rats and mice experienced significantly more episodes of bradypnea, indicative of impaired respiratory drive, after opioid microinjection. During spontaneous arousals, rats showed less tachycardia after opioid microinjection than before microinjection, suggestive of an attenuated sympathetic tone. Thus, activation of opioidergic signaling within RM produces effects beyond analgesia, including the unwanted destabilization of cardiorespiratory function. These adverse effects on homeostasis consequent to opioid microinjection imply a role for RM in regulating the balance of sympathetic and parasympathetic tone.

  14. A pilot study on predictors of brainstem raphe abnormality in patients with major depressive disorder.

    Science.gov (United States)

    Kostić, Milutin; Munjiza, Ana; Pesic, Danilo; Peljto, Amir; Novakovic, Ivana; Dobricic, Valerija; Tosevski, Dusica Lecic; Mijajlovic, Milija

    2017-02-01

    Hypo/anechogenicity of the brainstem raphe (BR) structures has been suggested as a possible transcranial parenchymal sonography (TCS) marker associated with depression. The aim of this study was to analyze possible association of the abnormal BR echogenicity in patients with major depression when compared to healthy controls, and to evaluate its clinical and genetic correlates. TCS was performed in 53 patients diagnosed as major depressive disorder (MDD) without psychotic symptoms and in 54 healthy matched controls. The TCS detected BR abnormalities were significantly more frequent in MDD patients (35 out of 53; 66%) in comparison to matched controls (5 out of 56; 9%). The prevalence of short allele (s) homozygocity in the length polymorphism of the promoter region of the serotonin transporter gene (5-HTTLPR) was significantly higher in MDD patients relative to those with normal BR echogenicity. A stepwise statistical discriminant analysis revealed statistically significant separation between MDD patients with and without BR abnormalities groups based on the four predictors combined: the Hamilton Anxiety Rating Scale item 5 ("difficulty in concentration, poor memory"), presence of social phobia, s allele homozygocity of the 5-HTTLPR polymorphism, and presence of generalized anxiety disorder. Cross-sectional design and heterogenous treatment of depressed patients. Reduced BR echogenicity in at least a subgroup of MDD patients may reflect a particular phenotype, characterized by more prevalent comorbid anxiety disorders, associated with particular genetic polymorphisms and neurotransmitter(s) deficits, most probably altered serotonergic mechanisms. Copyright © 2016 Elsevier B.V. All rights reserved.

  15. LHC Create

    CERN Multimedia

    CERN. Geneva

    2017-01-01

    LHC Create is an upcoming 2-day workshop held at IdeaSquare in November. Participants from CERN and IPAC school of design will compete to design an exhibit that explains why CERN does what it does. The winner will have their exhibit fully realised and made available to experiments, institutes, and tourism agencies around the world.

  16. Creating Poetry.

    Science.gov (United States)

    Drury, John

    Encouraging exploration and practice, this book offers hundreds of exercises and numerous tips covering every step involved in creating poetry. Each chapter is a self-contained unit offering an overview of material in the chapter, a definition of terms, and poetry examples from well-known authors designed to supplement the numerous exercises.…

  17. Francis Bacon's Valerius Terminus and the Voyage to the "Great Instauration".

    Science.gov (United States)

    Serjeantson, Richard

    2017-01-01

    Francis Bacon's earliest surviving natural philosophical treatise (composed circa 1603) bears the title Valerius Terminus of the Interpretation of Nature. This study, resting on fresh attention to the surviving authorial manuscript, has three goals. It begins by identifying a lost precursor work apparently entitled "Of Active Knowledge." It then examines the significance of the pseudonyms Bacon chose to introduce his ideas, considering especially his invocation of Erasmus's emblem, the Roman deity Terminus. Finally, it shows how the Valerius Terminus's global vision of contemporary knowledge ultimately helped shape the iconography of Bacon's published Instauratio magna.

  18. A novel fragile X syndrome mutation reveals a conserved role for the carboxy-terminus in FMRP localization and function.

    Science.gov (United States)

    Okray, Zeynep; de Esch, Celine E F; Van Esch, Hilde; Devriendt, Koen; Claeys, Annelies; Yan, Jiekun; Verbeeck, Jelle; Froyen, Guy; Willemsen, Rob; de Vrij, Femke M S; Hassan, Bassem A

    2015-04-01

    Loss of function of the FMR1 gene leads to fragile X syndrome (FXS), the most common form of intellectual disability. The loss of FMR1 function is usually caused by epigenetic silencing of the FMR1 promoter leading to expansion and subsequent methylation of a CGG repeat in the 5' untranslated region. Very few coding sequence variations have been experimentally characterized and shown to be causal to the disease. Here, we describe a novel FMR1 mutation and reveal an unexpected nuclear export function for the C-terminus of FMRP. We screened a cohort of patients with typical FXS symptoms who tested negative for CGG repeat expansion in the FMR1 locus. In one patient, we identified a guanine insertion in FMR1 exon 15. This mutation alters the open reading frame creating a short novel C-terminal sequence, followed by a stop codon. We find that this novel peptide encodes a functional nuclear localization signal (NLS) targeting the patient FMRP to the nucleolus in human cells. We also reveal an evolutionarily conserved nuclear export function associated with the endogenous C-terminus of FMRP. In vivo analyses in Drosophila demonstrate that a patient-mimetic mutation alters the localization and function of Dfmrp in neurons, leading to neomorphic neuronal phenotypes. © 2015 The Authors. Published under the terms of the CC BY 4.0 license.

  19. Hypothalamic projections to the ventral medulla oblongata in the rat, with special reference to the nucleus raphe pallidus: a study using autoradiographic and HRP techniques

    Energy Technology Data Exchange (ETDEWEB)

    Hosoya, Yasuhiko

    1985-10-07

    Hypothalamic descending projections to the medullary ventral surface were studied autoradiographically in the rat. A small amount of (/sup 3/H)leucine was injected unilaterally into various parts of the hypothalamus by air pressure. Abundant and characteristic terminal labelings were observed bilaterally in the nucleus raphe pallidus, the ventral surface to the pyramidal tract and the nucleus interfascicularis hypoglossi, after injections into the dorsal posterior hypothalamic area caudal to the paraventricular hypothalamic nucleus. Conspicuous, but less numerous labelings were observed in the nucleus raphe obscurus and the ipsilateral raphe magnus. After an injection of (/sup 3/H)leucine into the hypothalamus and injections of horseradish peroxidase (HRP) into the spinal cord in the same animal, silver grains were densely distributed around HRP-labeled neurons in the nucleus raphe pallidus including the nucleus interfascicularis hypoglossi. The present results suggest that the dorsal posterior hypothalamic area projects directly to the spinal-projecting neurons of the nucleus raphe pallidus. 53 refs.; 9 figs.

  20. Optogenetic activation of dorsal raphe serotonin neurons enhances patience for future rewards.

    Science.gov (United States)

    Miyazaki, Kayoko W; Miyazaki, Katsuhiko; Tanaka, Kenji F; Yamanaka, Akihiro; Takahashi, Aki; Tabuchi, Sawako; Doya, Kenji

    2014-09-08

    Serotonin is a neuromodulator that is involved extensively in behavioral, affective, and cognitive functions in the brain. Previous recording studies of the midbrain dorsal raphe nucleus (DRN) revealed that the activation of putative serotonin neurons correlates with the levels of behavioral arousal [1], rhythmic motor outputs [2], salient sensory stimuli [3-6], reward, and conditioned cues [5-8]. The classic theory on serotonin states that it opposes dopamine and inhibits behaviors when aversive events are predicted [9-14]. However, the therapeutic effects of serotonin signal-enhancing medications have been difficult to reconcile with this theory [15, 16]. In contrast, a more recent theory states that serotonin facilitates long-term optimal behaviors and suppresses impulsive behaviors [17-21]. To test these theories, we developed optogenetic mice that selectively express channelrhodopsin in serotonin neurons and tested how the activation of serotonergic neurons in the DRN affects animal behavior during a delayed reward task. The activation of serotonin neurons reduced the premature cessation of waiting for conditioned cues and food rewards. In reward omission trials, serotonin neuron stimulation prolonged the time animals spent waiting. This effect was observed specifically when the animal was engaged in deciding whether to keep waiting and was not due to motor inhibition. Control experiments showed that the prolonged waiting times observed with optogenetic stimulation were not due to behavioral inhibition or the reinforcing effects of serotonergic activation. These results show, for the first time, that the timed activation of serotonin neurons during waiting promotes animals' patience to wait for a delayed reward. Copyright © 2014 Elsevier Ltd. All rights reserved.

  1. Tolerance to Non-Opioid Analgesics is Opioid Sensitive in the Nucleus Raphe Magnus.

    Science.gov (United States)

    Tsagareli, Merab G; Nozadze, Ivliane; Tsiklauri, Nana; Gurtskaia, Gulnaz

    2011-01-01

    Repeated injection of opioid analgesics can lead to a progressive loss of effect. This phenomenon is known as tolerance. Several lines of investigations have shown that systemic, intraperitoneal administration or the microinjection of non-opioid analgesics, non-steroidal anti-inflammatory drugs (NSAIDs) into the midbrain periaqueductal gray matter induces antinociception with some effects of tolerance. Our recent study has revealed that microinjection of three drugs analgin, ketorolac, and xefocam into the central nucleus of amygdala produce tolerance to them and cross-tolerance to morphine. Here we report that repeated administrations of these NSAIDs into the nucleus raphe magnus (NRM) in the following 4 days result in progressively less antinociception compare to the saline control, i.e., tolerance develops to these drugs in male rats. Special control experiments showed that post-treatment with the μ-opioid antagonist naloxone into the NRM significantly decreased antinociceptive effects of NSAIDs on the first day of testing in the tail-flick (TF) reflex and hot plate (HP) latency tests. On the second day, naloxone generally had trend effects in both TF and HP tests and impeded the development of tolerance to the antinociceptive effect of non-opioid analgesics. These findings strongly support the suggestion of endogenous opioid involvement in NSAIDs antinociception and tolerance in the descending pain-control system. Moreover, repeated injections of NSAIDs progressively lead to tolerance to them, cross-tolerance to morphine, and the risk of a withdrawal syndrome. Therefore, these results are important for human medicine too.

  2. The role of the dorsal raphé nucleus in reward-seeking behavior

    Directory of Open Access Journals (Sweden)

    Kae eNakamura

    2013-08-01

    Full Text Available Pharmacological experiments have shown that the modulation of brain serotonin levels has a strong impact on value-based decision making. Anatomical and physiological evidence also revealed that the dorsal raphé nucleus (DRN, a major source of serotonin, and the dopamine system receive common inputs from brain regions associated with appetitive and aversive information processing. The serotonin and dopamine systems also have reciprocal functional influences on each other. However, the specific mechanism by which serotonin affects value-based decision making is not clear.To understand the information carried by the DRN for reward-seeking behavior, we measured single neuron activity in the primate DRN during the performance of saccade tasks to obtain different amounts of a reward. We found that DRN neuronal activity was characterized by tonic modulation that was altered by the expected and received reward value. Consistent reward-dependent modulation across different task periods suggested that DRN activity kept track of the reward value throughout a trial. The DRN was also characterized by modulation of its activity in the opposite direction by different neuronal subgroups, one firing strongly for the prediction and receipt of large rewards, with the other firing strongly for small rewards. Conversely, putative dopamine neurons showed positive phasic responses to reward-indicating cues and the receipt of an unexpected reward amount, which supports the reward prediction error signal hypothesis of dopamine.I suggest that the tonic reward monitoring signal of the DRN, possibly together with its interaction with the dopamine system, reports a continuous level of motivation throughout the performance of a task. Such a signal may provide reward context information to the targets of DRN projections, where it may be integrated further with incoming motivationally salient information.

  3. Chronic excitotoxic lesion of the dorsal raphe nucleus induces sodium appetite

    Directory of Open Access Journals (Sweden)

    Cavalcante-Lima H.R.

    2005-01-01

    Full Text Available We determined if the dorsal raphe nucleus (DRN exerts tonic control of basal and stimulated sodium and water intake. Male Wistar rats weighing 300-350 g were microinjected with phosphate buffer (PB-DRN, N = 11 or 1 µg/0.2 µl, in a single dose, ibotenic acid (IBO-DRN, N = 9 to 10 through a guide cannula into the DRN and were observed for 21 days in order to measure basal sodium appetite and water intake and in the following situations: furosemide-induced sodium depletion (20 mg/kg, sc, 24 h before the experiment and a low dose of dietary captopril (1 mg/g chow. From the 6th day after ibotenic acid injection IBO-DRN rats showed an increase in sodium appetite (12.0 ± 2.3 to 22.3 ± 4.6 ml 0.3 M NaCl intake whereas PB-DRN did not exceed 2 ml (P < 0.001. Water intake was comparable in both groups. In addition to a higher dipsogenic response, sodium-depleted IBO-DRN animals displayed an increase of 0.3 M NaCl intake compared to PB-DRN (37.4 ± 3.8 vs 21.6 ± 3.9 ml 300 min after fluid offer, P < 0.001. Captopril added to chow caused an increase of 0.3 M NaCl intake during the first 2 days (IBO-DRN, 33.8 ± 4.3 and 32.5 ± 3.4 ml on day 1 and day 2, respectively, vs 20.2 ± 2.8 ml on day 0, P < 0.001. These data support the view that DRN, probably via ascending serotonergic system, tonically modulates sodium appetite under basal and sodium depletion conditions and/or after an increase in peripheral or brain angiotensin II.

  4. Impacts of brain serotonin deficiency following Tph2 inactivation on development and raphe neuron serotonergic specification.

    Directory of Open Access Journals (Sweden)

    Lise Gutknecht

    Full Text Available Brain serotonin (5-HT is implicated in a wide range of functions from basic physiological mechanisms to complex behaviors, including neuropsychiatric conditions, as well as in developmental processes. Increasing evidence links 5-HT signaling alterations during development to emotional dysregulation and psychopathology in adult age. To further analyze the importance of brain 5-HT in somatic and brain development and function, and more specifically differentiation and specification of the serotonergic system itself, we generated a mouse model with brain-specific 5-HT deficiency resulting from a genetically driven constitutive inactivation of neuronal tryptophan hydroxylase-2 (Tph2. Tph2 inactivation (Tph2-/- resulted in brain 5-HT deficiency leading to growth retardation and persistent leanness, whereas a sex- and age-dependent increase in body weight was observed in Tph2+/- mice. The conserved expression pattern of the 5-HT neuron-specific markers (except Tph2 and 5-HT demonstrates that brain 5-HT synthesis is not a prerequisite for the proliferation, differentiation and survival of raphe neurons subjected to the developmental program of serotonergic specification. Furthermore, although these neurons are unable to synthesize 5-HT from the precursor tryptophan, they still display electrophysiological properties characteristic of 5-HT neurons. Moreover, 5-HT deficiency induces an up-regulation of 5-HT(1A and 5-HT(1B receptors across brain regions as well as a reduction of norepinephrine concentrations accompanied by a reduced number of noradrenergic neurons. Together, our results characterize developmental, neurochemical, neurobiological and electrophysiological consequences of brain-specific 5-HT deficiency, reveal a dual dose-dependent role of 5-HT in body weight regulation and show that differentiation of serotonergic neuron phenotype is independent from endogenous 5-HT synthesis.

  5. Neurochemical differences between target-specific populations of rat dorsal raphe projection neurons.

    Science.gov (United States)

    Prouty, Eric W; Chandler, Daniel J; Waterhouse, Barry D

    2017-11-15

    Serotonin (5-HT)-containing neurons in the dorsal raphe (DR) nucleus project throughout the forebrain and are implicated in many physiological processes and neuropsychiatric disorders. Diversity among these neurons has been characterized in terms of their neurochemistry and anatomical organization, but a clear sense of whether these attributes align with specific brain functions or terminal fields is lacking. DR 5-HT neurons can co-express additional neuroactive substances, increasing the potential for individualized regulation of target circuits. The goal of this study was to link DR neurons to a specific functional role by characterizing cells according to both their neurotransmitter expression and efferent connectivity; specifically, cells projecting to the medial prefrontal cortex (mPFC), a region implicated in cognition, emotion, and responses to stress. Following retrograde tracer injection, brainstem sections from Sprague-Dawley rats were immunohistochemically stained for markers of serotonin, glutamate, GABA, and nitric oxide (NO). 98% of the mPFC-projecting serotonergic neurons co-expressed the marker for glutamate, while the markers for NO and GABA were observed in 60% and less than 1% of those neurons, respectively. To identify potential target-specific differences in co-transmitter expression, we also characterized DR neurons projecting to a visual sensory structure, the lateral geniculate nucleus (LGN). The proportion of serotonergic neurons co-expressing NO was greater amongst cells targeting the mPFC vs LGN (60% vs 22%). The established role of 5-HT in affective disorders and the emerging role of NO in stress signaling suggest that the impact of 5-HT/NO co-localization in DR neurons that regulate mPFC circuit function may be clinically relevant. Copyright © 2017 Elsevier B.V. All rights reserved.

  6. A GIS tool for two-dimensional glacier-terminus change tracking

    Science.gov (United States)

    Urbanski, Jacek Andrzej

    2018-02-01

    This paper presents a Glacier Termini Tracking (GTT) toolbox for the two-dimensional analysis of glacier-terminus position changes. The input consists of a vector layer with several termini lines relating to the same glacier at different times. The output layers allow analyses to be conducted of glacier-terminus retreats, changes in retreats over time and along the ice face, and glacier-terminus fluctuations over time. The application of three tools from the toolbox is demonstrated via the analysis of eight glacier-terminus retreats and fluctuations at the Hornsund fjord in south Svalbard. It is proposed that this toolbox may also be useful in the study of other line features that change over time, like coastlines and rivers. The toolbox has been coded in Python and runs via ArcGIS.

  7. Division-induced DNA double strand breaks in the chromosome terminus region of Escherichia coli lacking RecBCD DNA repair enzyme.

    Directory of Open Access Journals (Sweden)

    Anurag Kumar Sinha

    2017-10-01

    Full Text Available Marker frequency analysis of the Escherichia coli recB mutant chromosome has revealed a deficit of DNA in a specific zone of the terminus, centred on the dif/TerC region. Using fluorescence microscopy of a marked chromosomal site, we show that the dif region is lost after replication completion, at the time of cell division, in one daughter cell only, and that the phenomenon is transmitted to progeny. Analysis by marker frequency and microscopy shows that the position of DNA loss is not defined by the replication fork merging point since it still occurs in the dif/TerC region when the replication fork trap is displaced in strains harbouring ectopic Ter sites. Terminus DNA loss in the recB mutant is also independent of dimer resolution by XerCD at dif and of Topo IV action close to dif. It occurs in the terminus region, at the point of inversion of the GC skew, which is also the point of convergence of specific sequence motifs like KOPS and Chi sites, regardless of whether the convergence of GC skew is at dif (wild-type or a newly created sequence. In the absence of FtsK-driven DNA translocation, terminus DNA loss is less precisely targeted to the KOPS convergence sequence, but occurs at a similar frequency and follows the same pattern as in FtsK+ cells. Importantly, using ftsIts, ftsAts division mutants and cephalexin treated cells, we show that DNA loss of the dif region in the recB mutant is decreased by the inactivation of cell division. We propose that it results from septum-induced chromosome breakage, and largely contributes to the low viability of the recB mutant.

  8. Expression and purification of antimicrobial peptide adenoregulin with C-amidated terminus in Escherichia coli.

    Science.gov (United States)

    Cao, Wei; Zhou, Yuxun; Ma, Yushu; Luo, Qingping; Wei, Dongzhi

    2005-04-01

    Adenoregulin is a 33 amino acid antimicrobial peptide isolated from the skin of the arboreal frog Phyllomedusa bicolor. Natural adenoregulin is synthesized with an amidated valine residue at C-terminus and shows lethal effects against filamentous fungi, as well as a broad spectrum of pathogenic microorganisms. A synthetic gene for adenoregulin (ADR) with an additional amino acid glutamine at C-terminus was cloned into pET32a vector to allow expression of ADR as a Trx fusion protein in Escherichia coli BL21(DE3). The resulting expression level of the fusion protein could reach up to 20% of the total cell proteins. The fusion protein could be purified effectively by Ni2+-chelating chromatography. Released from the fusion protein by enterokinase cleavage and purified to homogeneity, the recombinant ADR displayed antimicrobial activity similar to that of the synthetic ADR reported earlier. Comparing the antimicrobial activities of the recombinant adenoregulin with C-amidated terminus to that without an amidated C-terminus, we found that the amide of glutamine at C-terminus of ADR improved its potency on certain microorganisms such as Tritirachium album and Saccharomyces cerevisiae.

  9. Effect of MDMA-Induced Axotomy on the Dorsal Raphe Forebrain Tract in Rats: An In Vivo Manganese-Enhanced Magnetic Resonance Imaging Study.

    Directory of Open Access Journals (Sweden)

    Chuang-Hsin Chiu

    Full Text Available 3,4-Methylenedioxymethamphetamine (MDMA, also known as "Ecstasy", is a common recreational drug of abuse. Several previous studies have attributed the central serotonergic neurotoxicity of MDMA to distal axotomy, since only fine serotonergic axons ascending from the raphe nucleus are lost without apparent damage to their cell bodies. However, this axotomy has never been visualized directly in vivo. The present study examined the axonal integrity of the efferent projections from the midbrain raphe nucleus after MDMA exposure using in vivo manganese-enhanced magnetic resonance imaging (MEMRI. Rats were injected subcutaneously six times with MDMA (5 mg/kg or saline once daily. Eight days after the last injection, manganese ions (Mn2+ were injected stereotactically into the raphe nucleus, and a series of MEMRI images was acquired over a period of 38 h to monitor the evolution of Mn2+-induced signal enhancement across the ventral tegmental area, the medial forebrain bundle (MFB, and the striatum. The MDMA-induced loss of serotonin transporters was clearly evidenced by immunohistological staining consistent with the Mn2+-induced signal enhancement observed across the MFB and striatum. MEMRI successfully revealed the disruption of the serotonergic raphe-striatal projections and the variable effect of MDMA on the kinetics of Mn2+ accumulation in the MFB and striatum.

  10. Upregulation of the dorsal raphe nucleus-prefrontal cortex serotonin system by chronic treatment with escitalopram in hyposerotonergic Wistar-Kyoto rats

    NARCIS (Netherlands)

    Yamada, Makiko; Kawahara, Yukie; Kaneko, Fumi; Kishikawa, Yuki; Sotogaku, Naoki; Poppinga, Wilfred J.; Folgering, Joost H. A.; Dremencov, Eliyahu; Kawahara, Hiroshi; Nishi, Akinori

    Wistar-Kyoto (WKY) rats are sensitive to chronic stressors and exhibit depression-like behavior. Dorsal raphe nucleus (DRN) serotonin (5-HT) neurons projecting to the prefrontal cortex (PFC) comprise the important neurocircuitry underlying the pathophysiology of depression. To evaluate the DRN-PFC

  11. Adolescence fluoxetine increases serotonergic activity in the raphe-hippocampus axis and improves depression-like behaviors in female rats that experienced neonatal maternal separation.

    Science.gov (United States)

    Yoo, Sang Bae; Kim, Bom-Taeck; Kim, Jin Young; Ryu, Vitaly; Kang, Dong-Won; Lee, Jong-Ho; Jahng, Jeong Won

    2013-06-01

    This study was conducted to examine if fluoxetine, a selective 5-hydroxytryptamine (5-HT) reuptake inhibitor, would reverse adverse behavioral effects of neonatal maternal separation in female rats. Sprague-Dawley pups were separated from dam daily for 3h during postnatal day (PND) 1-14 (maternal separation; MS) or left undisturbed (non-handled; NH). Female NH and MS pups received intraperitoneal injection of fluoxetine (10mg/kg) or vehicle daily from PND 35 until the end of the whole experimental period. Rats were either subjected to behavioral tests during PND 44-54, or sacrificed for neurochemical analyses during PND 43-45. Daily food intake and weight gain of both NH and MS pups were suppressed by fluoxetine, with greater effects in MS pups. MS experience increased immobility and decrease swimming in forced swim test. Swimming was increased, although immobility was not significantly decreased, in MS females by adolescence fluoxetine. However, adolescence fluoxetine increased immobility during forced swim test and decreased time spent in open arms during elevated plus maze test in NH females. Fluoxetine normalized MS-induced decrease of the raphe 5-HT levels and increased 5-HT metabolism in the hippocampus in MS females, and increased the hypothalamic 5-HT both in NH and MS. Fluoxetine decreased the raphe 5-HT and increased the plasma corticosterone in NH females. Results suggest that decreased 5-HTergic activity in the raphe nucleus is implicated in the pathophysiology of depression-like behaviors, and increased 5-HTergic activities in the raphe-hippocampus axis may be a part of anti-depressant efficacy of fluoxetine, in MS females. Also, an extra-hypothalamic 5-HTergic activity may contribute to the increased anorectic efficacy of fluoxetine in MS females. Additionally, decreased 5-HT in the raphe and elevated plasma corticosterone may be related with fluoxetine-induced depression- and/or anxiety-like behaviors in NH females. Copyright © 2012 Elsevier Ltd

  12. The p21 ras C-terminus is required for transformation and membrane association

    DEFF Research Database (Denmark)

    Willumsen, B M; Christensen, A; Hubbert, N L

    1984-01-01

    The Harvey murine sarcoma virus (Ha-MuSV) transforming gene, v-rasH, encodes a 21,000 molecular weight protein (p21) that is closely related to the p21 proteins encoded by the cellular transforming genes of the ras gene family. The primary translation product (prop21), which is found in the cytosol...... of these biochemical features of the protein, we have now studied a series of deletion mutants located at or near the C-terminus of the viral p21 protein. Our tissue culture studies indicate that amino acids located at or near the C-terminus are required for cellular transformation, membrane association and lipid...

  13. Bacillus subtilis SepF binds to the C-terminus of FtsZ.

    Science.gov (United States)

    Król, Ewa; van Kessel, Sebastiaan P; van Bezouwen, Laura S; Kumar, Neeraj; Boekema, Egbert J; Scheffers, Dirk-Jan

    2012-01-01

    Bacterial cell division is mediated by a multi-protein machine known as the "divisome", which assembles at the site of cell division. Formation of the divisome starts with the polymerization of the tubulin-like protein FtsZ into a ring, the Z-ring. Z-ring formation is under tight control to ensure bacteria divide at the right time and place. Several proteins bind to the Z-ring to mediate its membrane association and persistence throughout the division process. A conserved stretch of amino acids at the C-terminus of FtsZ appears to be involved in many interactions with other proteins. Here, we describe a novel pull-down assay to look for binding partners of the FtsZ C-terminus, using a HaloTag affinity tag fused to the C-terminal 69 amino acids of B. subtilis FtsZ. Using lysates of Escherichia coli overexpressing several B. subtilis cell division proteins as prey we show that the FtsZ C-terminus specifically pulls down SepF, but not EzrA or MinC, and that the interaction depends on a conserved 16 amino acid stretch at the extreme C-terminus. In a reverse pull-down SepF binds to full-length FtsZ but not to a FtsZΔC16 truncate or FtsZ with a mutation of a conserved proline in the C-terminus. We show that the FtsZ C-terminus is required for the formation of tubules from FtsZ polymers by SepF rings. An alanine-scan of the conserved 16 amino acid stretch shows that many mutations affect SepF binding. Combined with the observation that SepF also interacts with the C-terminus of E. coli FtsZ, which is not an in vivo binding partner, we propose that the secondary and tertiary structure of the FtsZ C-terminus, rather than specific amino acids, are recognized by SepF.

  14. Bacillus subtilis SepF binds to the C-terminus of FtsZ.

    Directory of Open Access Journals (Sweden)

    Ewa Król

    Full Text Available Bacterial cell division is mediated by a multi-protein machine known as the "divisome", which assembles at the site of cell division. Formation of the divisome starts with the polymerization of the tubulin-like protein FtsZ into a ring, the Z-ring. Z-ring formation is under tight control to ensure bacteria divide at the right time and place. Several proteins bind to the Z-ring to mediate its membrane association and persistence throughout the division process. A conserved stretch of amino acids at the C-terminus of FtsZ appears to be involved in many interactions with other proteins. Here, we describe a novel pull-down assay to look for binding partners of the FtsZ C-terminus, using a HaloTag affinity tag fused to the C-terminal 69 amino acids of B. subtilis FtsZ. Using lysates of Escherichia coli overexpressing several B. subtilis cell division proteins as prey we show that the FtsZ C-terminus specifically pulls down SepF, but not EzrA or MinC, and that the interaction depends on a conserved 16 amino acid stretch at the extreme C-terminus. In a reverse pull-down SepF binds to full-length FtsZ but not to a FtsZΔC16 truncate or FtsZ with a mutation of a conserved proline in the C-terminus. We show that the FtsZ C-terminus is required for the formation of tubules from FtsZ polymers by SepF rings. An alanine-scan of the conserved 16 amino acid stretch shows that many mutations affect SepF binding. Combined with the observation that SepF also interacts with the C-terminus of E. coli FtsZ, which is not an in vivo binding partner, we propose that the secondary and tertiary structure of the FtsZ C-terminus, rather than specific amino acids, are recognized by SepF.

  15. A description of the lumbar interfascial triangle and its relation with the lateral raphe: anatomical constituents of load transfer through the lateral margin of the thoracolumbar fascia

    Science.gov (United States)

    Schuenke, M D; Vleeming, A; Van Hoof, T; Willard, F H

    2012-01-01

    rib to the iliac crest. This triangle results in the unification of different fascial sheaths along the lateral border of the TLF, creating a ridged-union of dense connective tissue that has been termed the lateral raphe (Spine, 9,1984, 163). This triangle may function in the distribution of laterally mediated tension to balance different viscoelastic moduli, along either the middle or posterior layers of the TLF. PMID:22582887

  16. Organization of Functional Long-Range Circuits Controlling the Activity of Serotonergic Neurons in the Dorsal Raphe Nucleus

    Directory of Open Access Journals (Sweden)

    Li Zhou

    2017-03-01

    Full Text Available Serotonergic neurons play key roles in various biological processes. However, circuit mechanisms underlying tight control of serotonergic neurons remain largely unknown. Here, we systematically investigated the organization of long-range synaptic inputs to serotonergic neurons and GABAergic neurons in the dorsal raphe nucleus (DRN of mice with a combination of viral tracing, slice electrophysiological, and optogenetic techniques. We found that DRN serotonergic neurons and GABAergic neurons receive largely comparable synaptic inputs from six major upstream brain areas. Upon further analysis of the fine functional circuit structures, we found both bilateral and ipsilateral patterns of topographic connectivity in the DRN for the axons from different inputs. Moreover, the upstream brain areas were found to bidirectionally control the activity of DRN serotonergic neurons by recruiting feedforward inhibition or via a push-pull mechanism. Our study provides a framework for further deciphering the functional roles of long-range circuits controlling the activity of serotonergic neurons in the DRN.

  17. Distinct effect of orphanin FQ in nucleus raphe magnus and nucleus reticularis gigantocellularis on the rat tail flick reflex.

    Science.gov (United States)

    Yang, Z; Zhang, Y; Wu, G

    2001-06-22

    The aim of the present study is to investigate the effects of orphanin FQ (OFQ) microinjected into the nucleus raphe magnus (NRM) and the nucleus reticularis gigantocellularis (NGC) on pain modulation. The tail-flick latency (TFL) was used as a behavioral index of nociceptive responsiveness. The result showed microinjection of OFQ into the NRM significantly increased the TFL, whereas microinjection of OFQ into the NGC decreased the TFL, suggesting the analgesic effect of OFQ in the NRM and the hyperalgesic effect of OFQ in the NGC. As there are three classes of putative pain modulating neurons in the rostral ventromedial medulla (RVM), the hyperalgesic or analgesic effect of OFQ in the RVM might depend upon the different class of the neurons being acted.

  18. Phosphorylation of CaMKII in the rat dorsal raphe nucleus plays an important role in sleep-wake regulation.

    Science.gov (United States)

    Cui, Su-Ying; Li, Sheng-Jie; Cui, Xiang-Yu; Zhang, Xue-Qiong; Yu, Bin; Sheng, Zhao-Fu; Huang, Yuan-Li; Cao, Qing; Xu, Ya-Ping; Lin, Zhi-Ge; Yang, Guang; Song, Jin-Zhi; Ding, Hui; Wang, Zi-Jun; Zhang, Yong-He

    2016-02-01

    The Ca(2+) modulation in the dorsal raphe nucleus (DRN) plays an important role in sleep-wake regulation. Calmodulin-dependent kinase II (CaMKII) is an important signal-transducing molecule that is activated by Ca(2+) . This study investigated the effects of intracellular Ca(2+) /CaMKII signaling in the DRN on sleep-wake states in rats. Maximum and minimum CaMKII phosphorylation was detected at Zeitgeber time 21 (ZT 21; wakefulness state) and ZT 3 (sleep state), respectively, across the light-dark rhythm in the DRN in rats. Six-hour sleep deprivation significantly reduced CaMKII phosphorylation in the DRN. Microinjection of the CAMKII activation inhibitor KN-93 (5 or 10 nmol) into the DRN suppressed wakefulness and enhanced rapid-eye-movement sleep (REMS) and non-REM sleep (NREMS). Application of a high dose of KN-93 (10 nmol) increased slow-wave sleep (SWS) time, SWS bouts, the mean duration of SWS, the percentage of SWS relative to total sleep, and delta power density during NREMS. Microinjection of CaCl2 (50 nmol) in the DRN increased CaMKII phosphorylation and decreased NREMS, SWS, and REMS. KN-93 abolished the inhibitory effects of CaCl2 on NREMS, SWS, and REMS. These data indicate a novel wake-promoting and sleep-suppressing role for the Ca(2+) /CaMKII signaling pathway in DRN neurons. We propose that the intracellular Ca(2+) /CaMKII signaling in the dorsal raphe nucleus (DRN) plays wake-promoting and sleep-suppressing role in rats. Intra-DRN application of KN-93 (CaMKII activation inhibitor) suppressed wakefulness and enhanced rapid-eye-movement sleep (REMS) and non-REMS (NREMS). Intra-DRN application of CaCl2 attenuated REMS and NREMS. We think these findings should provide a novel cellular and molecular mechanism of sleep-wake regulation. © 2015 International Society for Neurochemistry.

  19. Modifications of the C terminus affect functionality and stability of yeast triacylglycerol lipase Tgl3p.

    Science.gov (United States)

    Koch, Barbara; Schmidt, Claudia; Ploier, Birgit; Daum, Günther

    2014-07-11

    Lipid droplets are specific organelles for the storage of triacylglycerols and steryl esters. They are surrounded by a phospholipid monolayer with a small but specific set of proteins embedded. Assembly and insertion of proteins into this surface membrane is an intriguing question of lipid droplet biology. To address this question we studied the topology of Tgl3p, the major triacylglycerol lipase of the yeast Saccharomyces cerevisiae, on lipid droplets. Employing the method of limited proteolysis of lipid droplet surface proteins, we found that the C terminus of Tgl3p faces the inside of the organelle, whereas the N terminus is exposed at the cytosolic side of lipid droplets. Detailed analysis of the C terminus revealed a stretch of seven amino acids that are critical for protein stability and functionality. The negative charge of two aspartate residues within this stretch is crucial for lipase activity of Tgl3p. A portion of Tgl3p, which is located to the endoplasmic reticulum, exhibits a different topology. In the phospholipid bilayer of the endoplasmic reticulum the C terminus faces the cytosol, which results in instability of the protein. Thus, the topology of Tgl3p is important for its function and strongly dependent on the membrane environment. © 2014 by The American Society for Biochemistry and Molecular Biology, Inc.

  20. The N-terminus of TDP-43 promotes its oligomerization and enhances DNA binding affinity

    Energy Technology Data Exchange (ETDEWEB)

    Chang, Chung-ke [Institute of Biomedical Sciences, Academia Sinica, Taipei 115, Taiwan (China); Wu, Tzong-Huah [Institute of Chemistry, Academia Sinica, Taipei 115, Taiwan (China); Chemical Biology and Molecular Biophysics Program, Taiwan International Graduate Program, Institute of Biochemistry, Academia Sinica, Taipei 115, Taiwan (China); Institute of Bioinformatics and Structural Biology, National Tsing Hua University, Hsinchu 300, Taiwan (China); Wu, Chu-Ya [Institute of Chemistry, Academia Sinica, Taipei 115, Taiwan (China); Graduate Institute of Engineering, National Taiwan University of Science and Technology, Taipei 106, Taiwan (China); Chiang, Ming-hui; Toh, Elsie Khai-Woon [Institute of Biomedical Sciences, Academia Sinica, Taipei 115, Taiwan (China); Hsu, Yin-Chih; Lin, Ku-Feng [Institute of Chemistry, Academia Sinica, Taipei 115, Taiwan (China); Liao, Yu-heng [Institute of Biomedical Sciences, Academia Sinica, Taipei 115, Taiwan (China); Huang, Tai-huang, E-mail: bmthh@gate.sinica.edu.tw [Institute of Biomedical Sciences, Academia Sinica, Taipei 115, Taiwan (China); Department of Physics, National Taiwan Normal University, Taipei 106, Taiwan (China); Huang, Joseph Jen-Tse, E-mail: jthuang@chem.sinica.edu.tw [Institute of Chemistry, Academia Sinica, Taipei 115, Taiwan (China)

    2012-08-24

    Highlights: Black-Right-Pointing-Pointer The N-terminus of TDP-43 contains an independently folded structural domain (NTD). Black-Right-Pointing-Pointer The structural domains of TDP-43 are arranged in a beads-on-a-string fashion. Black-Right-Pointing-Pointer The NTD promotes TDP-43 oligomerization in a concentration-dependent manner. Black-Right-Pointing-Pointer The NTD may assist nucleic acid-binding activity of TDP-43. -- Abstract: TDP-43 is a DNA/RNA-binding protein associated with different neurodegenerative diseases such as amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD-U). Here, the structural and physical properties of the N-terminus on TDP-43 have been carefully characterized through a combination of nuclear magnetic resonance (NMR), circular dichroism (CD) and fluorescence anisotropy studies. We demonstrate for the first time the importance of the N-terminus in promoting TDP-43 oligomerization and enhancing its DNA-binding affinity. An unidentified structural domain in the N-terminus is also disclosed. Our findings provide insights into the N-terminal domain function of TDP-43.

  1. Exposure to an open-field arena increases c-Fos expression in a subpopulation of neurons in the dorsal raphe nucleus, including neurons projecting to the basolateral amygdaloid complex

    DEFF Research Database (Denmark)

    Hale, M.W.; Hay-Schmidt, A.; Mikkelsen, J.D.

    2008-01-01

    Serotonergic systems in the dorsal raphe nucleus are thought to play an important role in the regulation of anxiety states. To investigate responses of neurons in the dorsal raphe nucleus to a mild anxiety-related stimulus, we exposed rats to an open-field, under low-light or high-light conditions....... Treatment effects on c-Fos expression in serotonergic and non-serotonergic cells in the midbrain raphe nuclei were determined 2 h following open-field exposure or home cage control (CO) conditions. Rats tested under both light conditions responded with increases in c-Fos expression in serotonergic neurons...... within subdivisions of the midbrain raphe nuclei compared with CO rats. However, the total numbers of serotonergic neurons involved were small suggesting that exposure to the open-field may affect a subpopulation of serotonergic neurons. To determine if exposure to the open-field activates a subset...

  2. Receptor⁻Receptor Interactions in Multiple 5-HT1A Heteroreceptor Complexes in Raphe-Hippocampal 5-HT Transmission and Their Relevance for Depression and Its Treatment.

    Science.gov (United States)

    Borroto-Escuela, Dasiel O; Narváez, Manuel; Ambrogini, Patrizia; Ferraro, Luca; Brito, Ismel; Romero-Fernandez, Wilber; Andrade-Talavera, Yuniesky; Flores-Burgess, Antonio; Millon, Carmelo; Gago, Belen; Narvaez, Jose Angel; Odagaki, Yuji; Palkovits, Miklos; Diaz-Cabiale, Zaida; Fuxe, Kjell

    2018-06-03

    Due to the binding to a number of proteins to the receptor protomers in receptor heteromers in the brain, the term "heteroreceptor complexes" was introduced. A number of serotonin 5-HT1A heteroreceptor complexes were recently found to be linked to the ascending 5-HT pathways known to have a significant role in depression. The 5-HT1A⁻FGFR1 heteroreceptor complexes were involved in synergistically enhancing neuroplasticity in the hippocampus and in the dorsal raphe 5-HT nerve cells. The 5-HT1A protomer significantly increased FGFR1 protomer signaling in wild-type rats. Disturbances in the 5-HT1A⁻FGFR1 heteroreceptor complexes in the raphe-hippocampal 5-HT system were found in a genetic rat model of depression (Flinders sensitive line (FSL) rats). Deficits in FSL rats were observed in the ability of combined FGFR1 and 5-HT1A agonist cotreatment to produce antidepressant-like effects. It may in part reflect a failure of FGFR1 treatment to uncouple the 5-HT1A postjunctional receptors and autoreceptors from the hippocampal and dorsal raphe GIRK channels, respectively. This may result in maintained inhibition of hippocampal pyramidal nerve cell and dorsal raphe 5-HT nerve cell firing. Also, 5-HT1A⁻5-HT2A isoreceptor complexes were recently demonstrated to exist in the hippocampus and limbic cortex. They may play a role in depression through an ability of 5-HT2A protomer signaling to inhibit the 5-HT1A protomer recognition and signaling. Finally, galanin (1⁻15) was reported to enhance the antidepressant effects of fluoxetine through the putative formation of GalR1⁻GalR2⁻5-HT1A heteroreceptor complexes. Taken together, these novel 5-HT1A receptor complexes offer new targets for treatment of depression.

  3. Receptor–Receptor Interactions in Multiple 5-HT1A Heteroreceptor Complexes in Raphe-Hippocampal 5-HT Transmission and Their Relevance for Depression and Its Treatment

    Directory of Open Access Journals (Sweden)

    Dasiel O. Borroto-Escuela

    2018-06-01

    Full Text Available Due to the binding to a number of proteins to the receptor protomers in receptor heteromers in the brain, the term “heteroreceptor complexes” was introduced. A number of serotonin 5-HT1A heteroreceptor complexes were recently found to be linked to the ascending 5-HT pathways known to have a significant role in depression. The 5-HT1A–FGFR1 heteroreceptor complexes were involved in synergistically enhancing neuroplasticity in the hippocampus and in the dorsal raphe 5-HT nerve cells. The 5-HT1A protomer significantly increased FGFR1 protomer signaling in wild-type rats. Disturbances in the 5-HT1A–FGFR1 heteroreceptor complexes in the raphe-hippocampal 5-HT system were found in a genetic rat model of depression (Flinders sensitive line (FSL rats. Deficits in FSL rats were observed in the ability of combined FGFR1 and 5-HT1A agonist cotreatment to produce antidepressant-like effects. It may in part reflect a failure of FGFR1 treatment to uncouple the 5-HT1A postjunctional receptors and autoreceptors from the hippocampal and dorsal raphe GIRK channels, respectively. This may result in maintained inhibition of hippocampal pyramidal nerve cell and dorsal raphe 5-HT nerve cell firing. Also, 5-HT1A–5-HT2A isoreceptor complexes were recently demonstrated to exist in the hippocampus and limbic cortex. They may play a role in depression through an ability of 5-HT2A protomer signaling to inhibit the 5-HT1A protomer recognition and signaling. Finally, galanin (1–15 was reported to enhance the antidepressant effects of fluoxetine through the putative formation of GalR1–GalR2–5-HT1A heteroreceptor complexes. Taken together, these novel 5-HT1A receptor complexes offer new targets for treatment of depression.

  4. Electrophysiological Assessment of Serotonin and GABA Neuron Function in the Dorsal Raphe during the Third Trimester Equivalent Developmental Period in Mice.

    Science.gov (United States)

    Morton, Russell A; Yanagawa, Yuchio; Valenzuela, C Fernando

    2015-01-01

    Alterations in the development of the serotonin system can have prolonged effects, including depression and anxiety disorders later in life. Serotonin axonal projections from the dorsal raphe undergo extensive refinement during the first 2 weeks of postnatal life in rodents (equivalent to the third trimester of human pregnancy). However, little is known about the functional properties of serotonin and GABA neurons in the dorsal raphe during this critical developmental period. We assessed the functional properties and synaptic connectivity of putative serotoninergic neurons and GABAergic neurons in the dorsal raphe during early [postnatal day (P) P5-P7] and late (P15-P17) stages of the third trimester equivalent period using electrophysiology. Our studies demonstrate that GABAergic neurons are hyperexcitable at P5-P7 relative to P15-P17. Furthermore, putative serotonin neurons exhibit an increase in both excitatory and GABAA receptor-mediated spontaneous postsynaptic currents during this developmental period. Our data suggest that GABAergic neurons and putative serotonin neurons undergo significant electrophysiological changes during neonatal development.

  5. Inter- and intracellular relationship of substance P-containing neurons with serotonin and GABA in the dorsal raphe nucleus: combination of autoradiographic and immunocytochemical techniques

    International Nuclear Information System (INIS)

    Magoul, R.; Onteniente, B.; Oblin, A.; Calas, A.

    1986-01-01

    Double-labeling experiments were performed at the electron microscopic level in the dorsal raphe nucleus of rat, in order to study the inter- and intracellular relationship of substance P with gamma-aminobutyric acid (GABA) and serotonin. Autoradiography for either [ 3 H]serotonin or [ 3 H]GABA was coupled, on the same tissue section, with peroxidase-antiperoxidase immunocytochemistry for substance P in colchicine-treated animals. Intercellular relationships were represented by synaptic contacts made by [ 3 H]serotonin-labeled terminals on substance P-containing somata and dendrites, and by substance P-containing terminals on [ 3 H]GABA-labeled cells. Intracellular relationships were suggested by the occurrence of the peptide within [ 3 H]serotonin-containing and [ 3 H]GABA-containing cell bodies and fibers. Doubly labeled varicosities of the two kinds were also observed in the supraependymal plexus adjacent to the dorsal raphe nucleus. The results demonstrated that, in addition to reciprocal synaptic interactions made by substance P with serotonin and GABA, the dorsal raphe nucleus is the site of intracellular relationships between the peptide and either the amine or the amino acid

  6. Cadherin-13 Deficiency Increases Dorsal Raphe 5-HT Neuron Density and Prefrontal Cortex Innervation in the Mouse Brain

    Directory of Open Access Journals (Sweden)

    Andrea Forero

    2017-09-01

    Full Text Available Background: During early prenatal stages of brain development, serotonin (5-HT-specific neurons migrate through somal translocation to form the raphe nuclei and subsequently begin to project to their target regions. The rostral cluster of cells, comprising the median and dorsal raphe (DR, innervates anterior regions of the brain, including the prefrontal cortex. Differential analysis of the mouse 5-HT system transcriptome identified enrichment of cell adhesion molecules in 5-HT neurons of the DR. One of these molecules, cadherin-13 (Cdh13 has been shown to play a role in cell migration, axon pathfinding, and synaptogenesis. This study aimed to investigate the contribution of Cdh13 to the development of the murine brain 5-HT system.Methods: For detection of Cdh13 and components of the 5-HT system at different embryonic developmental stages of the mouse brain, we employed immunofluorescence protocols and imaging techniques, including epifluorescence, confocal and structured illumination microscopy. The consequence of CDH13 loss-of-function mutations on brain 5-HT system development was explored in a mouse model of Cdh13 deficiency.Results: Our data show that in murine embryonic brain Cdh13 is strongly expressed on 5-HT specific neurons of the DR and in radial glial cells (RGCs, which are critically involved in regulation of neuronal migration. We observed that 5-HT neurons are intertwined with these RGCs, suggesting that these neurons undergo RGC-guided migration. Cdh13 is present at points of intersection between these two cell types. Compared to wildtype controls, Cdh13-deficient mice display increased cell densities in the DR at embryonic stages E13.5, E17.5, and adulthood, and higher serotonergic innervation of the prefrontal cortex at E17.5.Conclusion: Our findings provide evidence for a role of CDH13 in the development of the serotonergic system in early embryonic stages. Specifically, we indicate that Cdh13 deficiency affects the cell

  7. Structure of the human MLH1 N-terminus: implications for predisposition to Lynch syndrome

    International Nuclear Information System (INIS)

    Wu, Hong; Zeng, Hong; Lam, Robert; Tempel, Wolfram; Kerr, Iain D.; Min, Jinrong

    2015-01-01

    The crystal structure of the human MLH1 N-terminus is reported at 2.30 Å resolution. The overall structure is described along with an analysis of two clinically important mutations. Mismatch repair prevents the accumulation of erroneous insertions/deletions and non-Watson–Crick base pairs in the genome. Pathogenic mutations in the MLH1 gene are associated with a predisposition to Lynch and Turcot’s syndromes. Although genetic testing for these mutations is available, robust classification of variants requires strong clinical and functional support. Here, the first structure of the N-terminus of human MLH1, determined by X-ray crystallography, is described. The structure shares a high degree of similarity with previously determined prokaryotic MLH1 homologs; however, this structure affords a more accurate platform for the classification of MLH1 variants

  8. Structure of the human MLH1 N-terminus: implications for predisposition to Lynch syndrome

    Energy Technology Data Exchange (ETDEWEB)

    Wu, Hong; Zeng, Hong; Lam, Robert; Tempel, Wolfram [University of Toronto, 101 College Street, Toronto, ON M5G 1L7 (Canada); Kerr, Iain D., E-mail: ikerr@myriad.com [Myriad Genetic Laboratories Inc., 320 Wakara Way, Salt Lake City, UT 84108 (United States); Min, Jinrong, E-mail: ikerr@myriad.com [University of Toronto, 101 College Street, Toronto, ON M5G 1L7 (Canada); University of Toronto, Toronto, ON M5G 1L7 (Canada)

    2015-07-28

    The crystal structure of the human MLH1 N-terminus is reported at 2.30 Å resolution. The overall structure is described along with an analysis of two clinically important mutations. Mismatch repair prevents the accumulation of erroneous insertions/deletions and non-Watson–Crick base pairs in the genome. Pathogenic mutations in the MLH1 gene are associated with a predisposition to Lynch and Turcot’s syndromes. Although genetic testing for these mutations is available, robust classification of variants requires strong clinical and functional support. Here, the first structure of the N-terminus of human MLH1, determined by X-ray crystallography, is described. The structure shares a high degree of similarity with previously determined prokaryotic MLH1 homologs; however, this structure affords a more accurate platform for the classification of MLH1 variants.

  9. Paired dating of pith and outer edge (terminus) samples from prehispanic Caribbean wooden sculptures

    Science.gov (United States)

    Fiona Brock; Joanna Ostapkowicz; Christopher Bronk Ramsey; Alex Wiedenhoeft; Caroline. Cartwright

    2012-01-01

    Radiocarbon dating of historical and archaeological wood can be complicated, sometimes involving issues of “inbuilt” age in slow-growing woods, and/or the possibility of reuse or long delays between felling and use of the wood. Terminus dates can be provided by dating the sapwood, or the outermost edge of heartwood, while a date from the pith can give an indication of...

  10. Controls on the inland propagation of terminus-driven speedups at Helheim Glacier, SE Greenland

    Science.gov (United States)

    Kehrl, L. M.; Joughin, I. R.; Smith, B.

    2017-12-01

    Tidewater glaciers are very sensitive to changes in the stress balance near their termini. When submarine melt or iceberg calving reduce lateral or basal resistance near the terminus, the glacier typically must speed up to produce the additional longitudinal and lateral stress gradients necessary to restore stress balance. Once speedup near the terminus is initiated, it can propagate inland through longitudinal stress coupling, thinning-induced changes in the effective pressure, and/or a steepening of surface slopes. The controls on these processes and the timing and spatial extent of the inland response, however, remain poorly understood. In this study, we use a three-dimensional, Full Stokes model (Elmer/Ice) to investigate the effects of different ice rheology and basal sliding parameterizations on the inland propagation of speedups at Helheim Glacier, SE Greenland. Using satellite observations of terminus position, we force the model with the observed 3-km, 2013/14 retreat history and allow the model to evolve in response to this retreat. We run a set of simulations that vary the ice rheology (constant or spatially variable ice temperature) and basal sliding law (linear, nonlinear, and effective-pressure-dependent). Our results show that the choice of parameterizations affect the timing and spatial extent of the inland response, but that the range of acceptable parameters can be constrained by comparing the model results to satellite observations of surface velocity and elevation.

  11. Epigenetic regulation of dorsal raphe GABA(B1a) associated with isolation-induced abnormal responses to social stimulation in mice.

    Science.gov (United States)

    Araki, Ryota; Hiraki, Yosuke; Nishida, Shoji; Kuramoto, Nobuyuki; Matsumoto, Kinzo; Yabe, Takeshi

    2016-02-01

    In isolation-reared mice, social encounter stimulation induces locomotor hyperactivity and activation of the dorsal raphe nucleus (DRN), suggesting that dysregulation of dorsal raphe function may be involved in abnormal behaviors. In this study, we examined the involvement of dorsal raphe GABAergic dysregulation in the abnormal behaviors of isolation-reared mice. We also studied an epigenetic mechanism underlying abnormalities of the dorsal raphe GABAergic system. Both mRNA and protein levels of GABA(B1a), a GABA(B) receptor subunit, were increased in the DRN of isolation-reared mice, compared with these levels in group-reared mice. In contrast, mRNA levels for other GABAergic system-related genes (GABA(A) receptor α1, β2 and γ2 subunits, GABA(B) receptor 1b and 2 subunits, and glutamate decarboxylase 67 and 65) were unchanged. Intra-DRN microinjection of 0.06 nmol baclofen (a GABA(B) receptor agonist) exacerbated encounter-induced hyperactivity and aggressive behavior, while microinjection of 0.3 nmol phaclofen (a GABA(B) receptor antagonist) attenuated encounter-induced hyperactivity and aggressive behavior in isolation-reared mice. Furthermore, microinjection of 0.06 nmol baclofen elicited encounter-induced hyperactivity in group-reared mice. Neither baclofen nor phaclofen affected immobility time in the forced swim test and hyperactivity in a novel environment of isolation reared mice. Bisulfite sequence analyses revealed that the DNA methylation level of the CpG island around the transcription start site (TSS) of GABA(B1a) was decreased in the DRN of isolation-reared mice. Chromatin immunoprecipitation analysis showed that histone H3 was hyperacetylated around the TSS of GABA(B1a) in the DRN of isolation-reared mice. These findings indicate that an increase in dorsal raphe GABA(B1a) expression via epigenetic regulation is associated with abnormal responses to social stimulation such as encounter-induced hyperactivity and aggressive behavior in isolation

  12. Organization of Functional Long-Range Circuits Controlling the Activity of Serotonergic Neurons in the Dorsal Raphe Nucleus.

    Science.gov (United States)

    Zhou, Li; Liu, Ming-Zhe; Li, Qing; Deng, Juan; Mu, Di; Sun, Yan-Gang

    2017-03-21

    Serotonergic neurons play key roles in various biological processes. However, circuit mechanisms underlying tight control of serotonergic neurons remain largely unknown. Here, we systematically investigated the organization of long-range synaptic inputs to serotonergic neurons and GABAergic neurons in the dorsal raphe nucleus (DRN) of mice with a combination of viral tracing, slice electrophysiological, and optogenetic techniques. We found that DRN serotonergic neurons and GABAergic neurons receive largely comparable synaptic inputs from six major upstream brain areas. Upon further analysis of the fine functional circuit structures, we found both bilateral and ipsilateral patterns of topographic connectivity in the DRN for the axons from different inputs. Moreover, the upstream brain areas were found to bidirectionally control the activity of DRN serotonergic neurons by recruiting feedforward inhibition or via a push-pull mechanism. Our study provides a framework for further deciphering the functional roles of long-range circuits controlling the activity of serotonergic neurons in the DRN. Copyright © 2017 The Author(s). Published by Elsevier Inc. All rights reserved.

  13. Activation of GABAergic pathway by hypocretin in the median raphe nucleus (MRN) mediates stress-induced theta rhythm in rats.

    Science.gov (United States)

    Hsiao, Yi-Tse; Jou, Shuo-Bin; Yi, Pei-Lu; Chang, Fang-Chia

    2012-07-15

    The frequency of electroencephalograms (EEGs) is predominant in theta rhythm during stress (e.g., footshock) in rats. Median raphe nucleus (MRN) desynchronizes hippocampal theta waves via activation of GABAergic neurons in the medial septum-diagonal band of Broca (MS-DBB), a theta rhythm pacemaker. Increased hypocretin mediates stress responses in addition to the maintenance of wakefulness. Hypocretin receptors are abundant in the MRN, suggesting a possible role of hypocretin in modulating stress-induced theta rhythm. Our results indicated that the intensity of theta waves was enhanced by footshock and that a hypocretin receptor antagonist (TCS1102) suppressed the footshock-induced theta waves. Administration of hypocretin-1 (1 and 10 μg) and hypocretin-2 (10 μg) directly into the MRN simulated the effect of footshock and significantly increased theta waves. Co-administration of GABA(A) receptor antagonist, bicuculline, into the MRN blocked the increase of theta waves induced by hypocretins or footshock. These results suggested that stress enhances the release of hypocretins, activates GABAergic neurons in the MRN, blocks the ability of MRN to desynchronize theta waves, and subsequently increases the intensity of theta rhythm. Copyright © 2012 Elsevier B.V. All rights reserved.

  14. Adaptive Control of Dorsal Raphe by 5-HT4 in the Prefrontal Cortex Prevents Persistent Hypophagia following Stress

    Directory of Open Access Journals (Sweden)

    Alexandra Jean

    2017-10-01

    Full Text Available Transient reduced food intake (hypophagia following high stress could have beneficial effects on longevity, but paradoxically, hypophagia can persist and become anorexia-like behavior. The neural underpinnings of stress-induced hypophagia and the mechanisms by which the brain prevents the transition from transient to persistent hypophagia remain undetermined. In this study, we report the involvement of a network governing goal-directed behavior (decision. This network consists of the ascending serotonergic inputs from the dorsal raphe nucleus (DR to the medial prefrontal cortex (mPFC. Specifically, adult restoration of serotonin 4 receptor (5-HT4R expression in the mPFC rescues hypophagia and specific molecular changes related to depression resistance in the DR (5-HT release elevation, 5-HT1A receptor, and 5-HT transporter reductions of stressed 5-HT4R knockout mice. The adult mPFC-5-HT4R knockdown mimics the null phenotypes. When mPFC-5-HT4Rs are overexpressed and DR-5-HT1ARs are blocked in the DR, hypophagia following stress persists, suggesting an antidepressant action of early anorexia.

  15. Adaptive Control of Dorsal Raphe by 5-HT4 in the Prefrontal Cortex Prevents Persistent Hypophagia following Stress.

    Science.gov (United States)

    Jean, Alexandra; Laurent, Laetitia; Delaunay, Sabira; Doly, Stéphane; Dusticier, Nicole; Linden, David; Neve, Rachael; Maroteaux, Luc; Nieoullon, André; Compan, Valérie

    2017-10-24

    Transient reduced food intake (hypophagia) following high stress could have beneficial effects on longevity, but paradoxically, hypophagia can persist and become anorexia-like behavior. The neural underpinnings of stress-induced hypophagia and the mechanisms by which the brain prevents the transition from transient to persistent hypophagia remain undetermined. In this study, we report the involvement of a network governing goal-directed behavior (decision). This network consists of the ascending serotonergic inputs from the dorsal raphe nucleus (DR) to the medial prefrontal cortex (mPFC). Specifically, adult restoration of serotonin 4 receptor (5-HT 4 R) expression in the mPFC rescues hypophagia and specific molecular changes related to depression resistance in the DR (5-HT release elevation, 5-HT 1A receptor, and 5-HT transporter reductions) of stressed 5-HT 4 R knockout mice. The adult mPFC-5-HT 4 R knockdown mimics the null phenotypes. When mPFC-5-HT 4 Rs are overexpressed and DR-5-HT1ARs are blocked in the DR, hypophagia following stress persists, suggesting an antidepressant action of early anorexia. Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.

  16. Serotonin neurons in the dorsal raphe mediate the anticataplectic action of orexin neurons by reducing amygdala activity.

    Science.gov (United States)

    Hasegawa, Emi; Maejima, Takashi; Yoshida, Takayuki; Masseck, Olivia A; Herlitze, Stefan; Yoshioka, Mitsuhiro; Sakurai, Takeshi; Mieda, Michihiro

    2017-04-25

    Narcolepsy is a sleep disorder caused by the loss of orexin (hypocretin)-producing neurons and marked by excessive daytime sleepiness and a sudden weakening of muscle tone, or cataplexy, often triggered by strong emotions. In a mouse model for narcolepsy, we previously demonstrated that serotonin neurons of the dorsal raphe nucleus (DRN) mediate the suppression of cataplexy-like episodes (CLEs) by orexin neurons. Using an optogenetic tool, in this paper we show that the acute activation of DRN serotonin neuron terminals in the amygdala, but not in nuclei involved in regulating rapid eye-movement sleep and atonia, suppressed CLEs. Not only did stimulating serotonin nerve terminals reduce amygdala activity, but the chemogenetic inhibition of the amygdala using designer receptors exclusively activated by designer drugs also drastically decreased CLEs, whereas chemogenetic activation increased them. Moreover, the optogenetic inhibition of serotonin nerve terminals in the amygdala blocked the anticataplectic effects of orexin signaling in DRN serotonin neurons. Taken together, the results suggest that DRN serotonin neurons, as a downstream target of orexin neurons, inhibit cataplexy by reducing the activity of amygdala as a center for emotional processing.

  17. Ribosomal DNA transcription in the dorsal raphe nucleus is increased in residual but not in paranoid schizophrenia.

    Science.gov (United States)

    Krzyżanowska, Marta; Steiner, Johann; Brisch, Ralf; Mawrin, Christian; Busse, Stefan; Braun, Katharina; Jankowski, Zbigniew; Bernstein, Hans-Gert; Bogerts, Bernhard; Gos, Tomasz

    2015-03-01

    The central serotonergic system is implicated in the pathogenesis of schizophrenia, where the imbalance between dopamine, serotonin and glutamate plays a key pathophysiological role. The dorsal raphe nucleus (DRN) is the main source of serotonergic innervation of forebrain limbic structures disturbed in schizophrenia patients. The study was carried out on paraffin-embedded brains from 17 (8 paranoid and 9 residual) schizophrenia patients and 28 matched controls without mental disorders. The transcriptional activity of ribosomal DNA (rDNA) in DRN neurons was evaluated by the AgNOR silver-staining method. An increased rDNA transcriptional activity was found in schizophrenia patients in the cumulative analysis of all DRN subnuclei (t test, P = 0.02). Further subgroup analysis revealed that it was an effect specific for residual schizophrenia versus paranoid schizophrenia or control groups (ANOVA, P = 0.002). This effect was confounded neither by suicide nor by antipsychotic medication. Our findings suggest that increased activity of rDNA in DRN neurons is a distinct phenomenon in schizophrenia, particularly in residual patients. An activation of the rDNA transcription in DRN neurons may represent a compensatory mechanism to overcome the previously described prefrontal serotonergic hypofunction in this diagnostic subgroup.

  18. Effect of acupuncture on Lipopolysaccharide-induced anxiety-like behavioral changes: involvement of serotonin system in dorsal Raphe nucleus.

    Science.gov (United States)

    Yang, Tae Young; Jang, Eun Young; Ryu, Yeonhee; Lee, Gyu Won; Lee, Eun Byeol; Chang, Suchan; Lee, Jong Han; Koo, Jin Suk; Yang, Chae Ha; Kim, Hee Young

    2017-12-11

    Acupuncture has been used as a common therapeutic tool in many disorders including anxiety and depression. Serotonin transporter (SERT) plays an important role in the pathology of anxiety and other mood disorders. The aim of this study was to evaluate the effects of acupuncture on lipopolysaccharide (LPS)-induced anxiety-like behaviors and SERT in the dorsal raphe nuclei (DRN). Rats were given acupuncture at ST41 (Jiexi), LI11 (Quchi) or SI3 (Houxi) acupoint in LPS-treated rats. Anxiety-like behaviors of elevated plus maze (EPM) and open field test (OFT) were measured and expressions of SERT and/or c-Fos were also examined in the DRN using immunohistochemistry. The results showed that 1) acupuncture at ST41 acupoint, but neither LI11 nor SI3, significantly attenuated LPS-induced anxiety-like behaviors in EPM and OFT, 2) acupuncture at ST41 decreased SERT expression increased by LPS in the DRN. Our results suggest that acupuncture can ameliorate anxiety-like behaviors, possibly through regulation of SERT in the DRN.

  19. A highly conserved glycine within linker I and the extreme C terminus of G protein alpha subunits interact cooperatively in switching G protein-coupled receptor-to-effector specificity

    DEFF Research Database (Denmark)

    Kostenis, Evi; Martini, Lene; Ellis, James

    2004-01-01

    Numerous studies have attested to the importance of the extreme C terminus of G protein alpha subunits in determining their selectivity of receptor recognition. We have previously reported that a highly conserved glycine residue within linker I is important for constraining the fidelity of receptor...... recognition by Galpha(q) proteins. Herein, we explored whether both modules (linker I and extreme C terminus) interact cooperatively in switching G protein-coupled receptor (GPCR)-to-effector specificity and created as models mutant Galpha(q) proteins in which glycine was replaced with various amino acids...... and the C-terminal five Galpha(q) residues with the corresponding Galpha(i) or Galpha(s) sequence. Coupling properties of the mutated Galpha(q) proteins were determined after coexpression with a panel of 13 G(i)-and G(s) -selective receptors and compared with those of Galpha proteins modified in only one...

  20. C-terminus glycans with critical functional role in the maturation of secretory glycoproteins.

    Directory of Open Access Journals (Sweden)

    Daniela Cioaca

    Full Text Available The N-glycans of membrane glycoproteins are mainly exposed to the extracellular space. Human tyrosinase is a transmembrane glycoprotein with six or seven bulky N-glycans exposed towards the lumen of subcellular organelles. The central active site region of human tyrosinase is modeled here within less than 2.5 Å accuracy starting from Streptomyces castaneoglobisporus tyrosinase. The model accounts for the last five C-terminus glycosylation sites of which four are occupied and indicates that these cluster in two pairs--one in close vicinity to the active site and the other on the opposite side. We have analyzed and compared the roles of all tyrosinase N-glycans during tyrosinase processing with a special focus on the proximal to the active site N-glycans, s6:N337 and s7:N371, versus s3:N161 and s4:N230 which decorate the opposite side of the domain. To this end, we have constructed mutants of human tyrosinase in which its seven N-glycosylation sites were deleted. Ablation of the s6:N337 and s7:N371 sites arrests the post-translational productive folding process resulting in terminally misfolded mutants subjected to degradation through the mannosidase driven ERAD pathway. In contrast, single mutants of the other five N-glycans located either opposite to the active site or into the N-terminus Cys1 extension of tyrosinase are temperature-sensitive mutants and recover enzymatic activity at the permissive temperature of 31°C. Sites s3 and s4 display selective calreticulin binding properties. The C-terminus sites s7 and s6 are critical for the endoplasmic reticulum retention and intracellular disposal. Results herein suggest that individual N-glycan location is critical for the stability, regional folding control and secretion of human tyrosinase and explains some tyrosinase gene missense mutations associated with oculocutaneous albinism type I.

  1. The role of the Cx43 C-terminus in GJ plaque formation and internalization

    International Nuclear Information System (INIS)

    Wayakanon, Praween; Bhattacharjee, Rajib; Nakahama, Ken-ichi; Morita, Ikuo

    2012-01-01

    Highlights: ► Cx43-GFP or -DsRed fusion proteins were expressed in HeLa cells. ► Roles of C-terminus were examined using various mutants. ► Gap junction plaque size was dependent on the length of C-terminus. ► C-terminus dependent gap junction plaque internalization was observed. -- Abstract: Connexin 43 (Cx43) is a major gap junction (GJ) protein found in many mammalian cell types. The C-terminal (CT) domain of Cx43 has unique characteristics in terms of amino acid (aa) sequence and its length differs from other connexins. This CT domain can be associated with protein partners to regulate GJ assembly and degradation, which results in the direct control of gap junction intercellular communication (GJIC). However, the essential roles of the CT regions involved in these mechanisms have not been fully elucidated. In this study, we aimed to investigate the specific regions of Cx43CT involved in GJ formation and internalization. Wild type Cx43 (382aa) and 10 CT truncated mutants were stably expressed in HeLa cells as GFP or DsRed tagged proteins. First, we found that the deletion of 235–382aa from Cx43 resulted in failure to make GJ and establish GJIC. Second, the Cx43 with 242–382aa CT deletion could form functional GJs and be internalized as annular gap junctions (AGJs). However, the plaques consisting of Cx43 with CT deletions (Δ242–382aa to Δ271–382aa) were longer than the plaques consisting of Cx43 with CT deletions (Δ302–382aa). Third, co-culture experiments of cells expressing wild type Cx43 (382) with cells expressing Cx43CT mutants revealed that the directions of GJ internalization were dependent on the length of the respective CT. Moreover, a specific region, 325–342aa residues of Cx43, played an important role in the direction of GJ internalization. These results showed the important roles of the Cx43 C-terminus in GJ expression and its turnover.

  2. The N terminus of monoamine transporters is a lever required for the action of amphetamines

    DEFF Research Database (Denmark)

    Sucic, Sonja; Dallinger, Stefan; Zdrazil, Barbara

    2010-01-01

    The serotonin transporter (SERT) terminates neurotransmission by removing serotonin from the synaptic cleft. In addition, it is the site of action of antidepressants (which block the transporter) and of amphetamines (which induce substrate efflux). We explored the functional importance of the N......(+) entry and accumulation of SERT(T81A) in the inward facing conformation ought to favor amphetamine-induced efflux. Thus, we surmised that the N terminus must play a direct role in driving the transporter into a state that supports amphetamine-induced efflux. This hypothesis was verified by truncating...

  3. Sex differences in corticotropin-releasing factor receptor-1 action within the dorsal raphe nucleus in stress responsivity.

    Science.gov (United States)

    Howerton, Alexis R; Roland, Alison V; Fluharty, Jessica M; Marshall, Anikò; Chen, Alon; Daniels, Derek; Beck, Sheryl G; Bale, Tracy L

    2014-06-01

    Women are twice as likely as men to suffer from stress-related affective disorders. Corticotropin-releasing factor (CRF) is an important link between stress and mood, in part through its signaling in the serotonergic dorsal raphe (DR). Development of CRF receptor-1 (CRFr1) antagonists has been a focus of numerous clinical trials but has not yet been proven efficacious. We hypothesized that sex differences in CRFr1 modulation of DR circuits might be key determinants in predicting therapeutic responses and affective disorder vulnerability. Male and female mice received DR infusions of the CRFr1 antagonist, NBI 35965, or CRF and were evaluated for stress responsivity. Sex differences in indices of neural activation (cFos) and colocalization of CRFr1 throughout the DR were examined. Whole-cell patch-clamp electrophysiology assessed sex differences in serotonin neuron membrane characteristics and responsivity to CRF. Males showed robust behavioral and hypothalamic-pituitary-adrenal axis responses to DR infusion of NBI 35965 and CRF, whereas females were minimally responsive. Sex differences were also found for both CRF-induced DR cFos and CRFr1 co-localization throughout the DR. Electrophysiologically, female serotonergic neurons showed blunted membrane excitability and divergent inhibitory postsynaptic current responses to CRF application. These studies demonstrate convincing sex differences in CRFr1 activity in the DR, where blunted female responses to NBI 35965 and CRF suggest unique stress modulation of the DR. These sex differences might underlie affective disorder vulnerability and differential sensitivity to pharmacologic treatments developed to target the CRF system, thereby contributing to a current lack of CRFr1 antagonist efficacy in clinical trials. © 2013 Published by Society of Biological Psychiatry on behalf of Society of Biological Psychiatry.

  4. Dorsal-to-Ventral Shift in Midbrain Dopaminergic Projections and Increased Thalamic/Raphe Serotonergic Function in Early Parkinson Disease.

    Science.gov (United States)

    Joutsa, Juho; Johansson, Jarkko; Seppänen, Marko; Noponen, Tommi; Kaasinen, Valtteri

    2015-07-01

    Loss of nigrostriatal neurons leading to dopamine depletion in the dorsal striatum is the pathologic hallmark of Parkinson disease contributing to the primary motor symptoms of the disease. However, Parkinson pathology is more widespread in the brain, affecting also other dopaminergic pathways and neurotransmitter systems, but these changes are less well characterized. This study aimed to investigate the mesencephalic striatal and extrastriatal dopaminergic projections together with extrastriatal serotonin transporter binding in Parkinson disease. Two hundred sixteen patients with Parkinson disease and 204 control patients (patients without neurodegenerative parkinsonism syndromes and normal SPECT imaging) were investigated with SPECT using the dopamine/serotonin transporter ligand (123)I-N-ω-fluoropropyl-2β-carbomethoxy-3β-(4-iodophenyl)nortropane ((123)I-FP-CIT) in the clinical setting. The group differences and midbrain correlations were analyzed voxel by voxel over the entire brain. We found that Parkinson patients had lower (123)I-FP-CIT uptake in the striatum and ventral midbrain but higher uptake in the thalamus and raphe nuclei than control patients. In patients with Parkinson disease, the correlation of the midbrain tracer uptake was shifted from the putamen to widespread corticolimbic areas. All findings were highly significant at the voxel level familywise error-corrected P value of less than 0.05. Our findings show that Parkinson disease is associated not only with the degeneration of the nigrostriatal dopamine neurotransmission, but also with a parallel shift toward mesolimbic and mesocortical function. Furthermore, Parkinson disease patients seem to have upregulation of brain serotonin transporter function at the early phase of the disease. © 2015 by the Society of Nuclear Medicine and Molecular Imaging, Inc.

  5. Optogenetic modulation of descending prefrontocortical inputs to the dorsal raphe bidirectionally bias socioaffective choices after social defeat

    Directory of Open Access Journals (Sweden)

    Collin eChallis

    2014-02-01

    Full Text Available It has been well established that modulating serotonin (5-HT levels in humans and animals affects perception and response to social threats, however the circuit mechanisms that control 5-HT output during social interaction are not well understood. A better understanding of these systems could provide groundwork for more precise and efficient therapeutic interventions. Here we examined the organization and plasticity of microcircuits implicated in top-down control of 5-HT neurons in the dorsal raphe nucleus (DRN by excitatory inputs from the ventromedial prefrontal cortex (vmPFC and their role in social approach-avoidance decisions. We did this in the context of a social defeat model that induces a long lasting form of social aversion that is reversible by antidepressants. We first used viral tracing and Cre-dependent genetic identification of vmPFC glutamatergic synapses in the DRN to determine their topographic distribution in relation to 5-HT and GABAergic subregions and found that excitatory vmPFC projections primarily localized to GABA-rich areas of the DRN. We then used optogenetics in combination with cFos mapping and slice electrophysiology to establish the functional effects of repeatedly driving vmPFC inputs in DRN. We provide the first direct evidence that vmPFC axons drive synaptic activity and immediate early gene expression in genetically identified DRN GABA neurons through an AMPA receptor-dependent mechanism. In contrast, we did not detect vmPFC-driven synaptic activity in 5-HT neurons and cFos induction in 5-HT neurons was limited. Finally we show that optogenetically increasing or decreasing excitatory vmPFC input to the DRN during sensory exposure to an aggressor’s cues enhances or diminishes avoidance bias, respectively. These results clarify the functional organization of vmPFC-DRN pathways and identify GABAergic neurons as a key cellular element filtering top-down vmPFC influences on affect-regulating 5-HT output.

  6. Effects of GABA microinjection into dorsal raphe nucleus on behavior and activity of lateral habenular neurons in mice.

    Science.gov (United States)

    Xiao, Jinyu; Song, Meiying; Li, Fengdan; Liu, Xiaofeng; Anwar, Alinur; Zhao, Hua

    2017-12-01

    The dorsal raphe nucleus (DRN) is a key site for 5-hydroxytryptamine (5-HT) synthesis and release. DRN dysfunction has been implicated in several stress-related disorders, including depression and anxiety. The lateral habenular nucleus (LHb) has been shown to inhibit the activity of DRN 5-HT neurons, and thus the LHb-DRN pathway plays an important role in the pathogenesis of depression. Although it is known that the LHb also receives the projection from the 5-HT neuron in the DRN, whether 5-HT neurons in the DRN can influence activity of the LHb in vivo and whether this effect is related to the induced behavioral changes have not been investigated. In the current study, we determined how injecting γ-aminobutyric acid (GABA) into the DRN to inhibit 5-HT neurons affected behavior and the changes in the activity of LHb neurons in mice. We found that GABA injection into the DRN induced depression-like behavior in mice, as indicated by increased immobility time, and decreased climbing time in the forced swimming test and the tail suspension test, decreased time spent in the center and total distance moved in the open field test. Using extracellular single unit recording, we showed that the firing rate of LHb neurons decreased after GABA microinjection into the DRN. Further, c-Fos expression in LHb neurons was inhibited. Together our results indicate that inhibition of DRN 5-HT neurons can cause decreased LHb activity and depression-like behavior in mice, however this depression-like behavior could be independent of the LHb activity. The observed decrease in LHb activity is probably due to the presence of a negative feedback loop between the DRN and the LHb, which may play a role in maintaining emotional homeostasis. Copyright © 2017 Elsevier Inc. All rights reserved.

  7. Ovarian steroid regulation of monoamine oxidase-A and -B mRNAs in the macaque dorsal raphe and hypothalamic nuclei.

    Science.gov (United States)

    Gundlah, Chrisana; Lu, Nick Z; Bethea, Cynthia L

    2002-03-01

    The serotonin neural system plays a pivotal role in mood, affective regulation and integrative cognition, as well as numerous autonomic functions. We have shown that ovarian steroids alter the expression of several genes in the dorsal raphe of macaques, which may increase serotonin synthesis and decrease serotonin autoinhibition. Another control point in aminergic neurotransmission involves degradation by MAO. This enzyme occurs in two isoforms, A and B, which have different substrate preferences. We questioned the effect of ovarian steroid hormones on MAO-A and MAO-B mRNA expression in the dorsal raphe nucleus and hypothalamus using in situ hybridization in non-human primates. Rhesus monkeys ( Macaca mulatta; n=5/group) were spayed and either placebo treated (controls), estrogen (E) treated (28 days), progesterone (P) treated (14 days placebo+14 days P), or E+P treated (14 days E+14 days E+P). Perfusion-fixed sections (25 microm) were hybridized with a 233 bp MAO-A, or a 373 bp MAO-B, radiolabeled-antisense monkey specific probes. Autoradiographic films were analyzed by densitometry, which was performed with NIH Image Software. MAO-A and -B mRNAs were detected in the dorsal raphe nucleus (DRN) and in the hypothalamic suprachiasmatic nucleus (SCN), preoptic area (POA), paraventricular nucleus (PVN), supraoptic nucleus (SON), lateral hypothalamus (LH) and ventromedial nucleus (VMN). MAO-A mRNA optical density was significantly decreased by E, P, and E+P in the DRN and in the hypothalamic PVN, LH and VMN. Ovarian hormones had no effect on MAO-B mRNA expression in the DRN. However, there was a significant decrease in MAO-B optical density in the hypothalamic POA, LH and VMN with E, P or E+P treatment. Pixel area generally reflected optical density. Ovarian steroids decreased MAO-A, but not B, in the raphe nucleus. However, both MAO-A and B were decreased in discrete hypothalamic nuclei by hormone replacement. These data suggest that the transcriptional regulation of

  8. Agonist-dependent modulation of G-protein coupling and transduction of 5-HT1A receptors in rat dorsal raphe nucleus

    OpenAIRE

    Valdizán, Elsa M.; Castro, Elena; Pazos, Ángel

    2009-01-01

    5-HT1A receptors couple to different Go/Gi proteins in order to mediate a wide range of physiological actions. While activation of post-synaptic 5-HT1A receptors is mainly related to inhibition of adenylyl cyclase activity, functionality of autoreceptors located in raphe nuclei has been classically ascribed to modifications of the activity of potassium and calcium channels. In order to evaluate the possible existence of agonist-directed trafficking for 5-HT1A autoreceptors in the rat dorsal r...

  9. MESOMARK kit detects C-ERC/mesothelin, but not SMRP with C-terminus.

    Science.gov (United States)

    Segawa, Tatsuya; Hagiwara, Yoshiaki; Ishikawa, Kiyoshi; Aoki, Naoko; Maeda, Masahiro; Shiomi, Kazu; Hino, Okio

    2008-05-09

    ERC/mesothelin is expressed on the normal mesothelium and some cancers such as mesothelioma or ovarian carcinoma. A splicing isoform of ERC/mesothelin (known as SMRP), which has an 82-bp insertion and codes for a C-terminus with a hydrophilic, presumably soluble, tail instead of a GPI-anchoring signal, has been reported as a useful marker for the diagnosis of mesothelioma. However, the existence of SMRP has not yet been demonstrated in the serum of mesothelioma patients. To elucidate the existence of SMRP, we have established a new enzyme-linked immunosorbent assay (ELISA) system for SMRP. The ELISA study revealed that N- and C-ERC/mesothelin were detected in sera from mesothelioma patients, but not SMRP, even in these samples. This result showed that the SMRP detected with MESOMARK kit should be lack of soluble C-terminus and indistinguishable from C-ERC/mesothelin. Further study might be necessary to demonstrate the relationship between SMRP and mesothelin.

  10. Capping of the N-terminus of PSD-95 by calmodulin triggers its postsynaptic release

    Science.gov (United States)

    Zhang, Yonghong; Matt, Lucas; Patriarchi, Tommaso; Malik, Zulfiqar A; Chowdhury, Dhrubajyoti; Park, Deborah K; Renieri, Alessandra; Ames, James B; Hell, Johannes W

    2014-01-01

    Postsynaptic density protein-95 (PSD-95) is a central element of the postsynaptic architecture of glutamatergic synapses. PSD-95 mediates postsynaptic localization of AMPA receptors and NMDA receptors and plays an important role in synaptic plasticity. PSD-95 is released from postsynaptic membranes in response to Ca2+ influx via NMDA receptors. Here, we show that Ca2+/calmodulin (CaM) binds at the N-terminus of PSD-95. Our NMR structure reveals that both lobes of CaM collapse onto a helical structure of PSD-95 formed at its N-terminus (residues 1–16). This N-terminal capping of PSD-95 by CaM blocks palmitoylation of C3 and C5, which is required for postsynaptic PSD-95 targeting and the binding of CDKL5, a kinase important for synapse stability. CaM forms extensive hydrophobic contacts with Y12 of PSD-95. The PSD-95 mutant Y12E strongly impairs binding to CaM and Ca2+-induced release of PSD-95 from the postsynaptic membrane in dendritic spines. Our data indicate that CaM binding to PSD-95 serves to block palmitoylation of PSD-95, which in turn promotes Ca2+-induced dissociation of PSD-95 from the postsynaptic membrane. PMID:24705785

  11. The N-terminus of survivin is a mitochondrial-targeting sequence and Src regulator

    Science.gov (United States)

    Dunajová, Lucia; Cash, Emily; Markus, Robert; Rochette, Sophie; Townley, Amelia R.

    2016-01-01

    ABSTRACT Survivin (also known as BIRC5) is a cancer-associated protein that exists in several locations in the cell. Its cytoplasmic residence in interphase cells is governed by CRM1 (also known as XPO1)-mediated nuclear exportation, and its localisation during mitosis to the centromeres and midzone microtubules is that of a canonical chromosomal passenger protein. In addition to these well-established locations, survivin is also a mitochondrial protein, but how it gets there and its function therein is presently unclear. Here, we show that the first ten amino acids at the N-terminus of survivin are sufficient to target GFP to the mitochondria in vivo, and ectopic expression of this decapeptide decreases cell adhesion and accelerates proliferation. The data support a signalling mechanism in which this decapeptide regulates the tyrosine kinase Src, leading to reduced focal adhesion plaques and disruption of F-actin organisation. This strongly suggests that the N-terminus of survivin is a mitochondrial-targeting sequence that regulates Src, and that survivin acts in concert with Src to promote tumorigenesis. PMID:27246243

  12. The Ku80 carboxy terminus stimulates joining and artemis-mediated processing of DNA ends

    DEFF Research Database (Denmark)

    Weterings, Eric; Verkaik, Nicole S; Keijzers, Guido

    2008-01-01

    Repair of DNA double-strand breaks (DSBs) is predominantly mediated by nonhomologous end joining (NHEJ) in mammalian cells. NHEJ requires binding of the Ku70-Ku80 heterodimer (Ku70/80) to the DNA ends and subsequent recruitment of the DNA-dependent protein kinase catalytic subunit (DNA-PK(CS)) an......Repair of DNA double-strand breaks (DSBs) is predominantly mediated by nonhomologous end joining (NHEJ) in mammalian cells. NHEJ requires binding of the Ku70-Ku80 heterodimer (Ku70/80) to the DNA ends and subsequent recruitment of the DNA-dependent protein kinase catalytic subunit (DNA......-PK(CS)) and the XRCC4/ligase IV complex. Activation of the DNA-PK(CS) serine/threonine kinase requires an interaction with Ku70/80 and is essential for NHEJ-mediated DSB repair. In contrast to previous models, we found that the carboxy terminus of Ku80 is not absolutely required for the recruitment and activation...... was phosphorylated to normal levels. This resulted in severely reduced levels of Artemis nuclease activity in vivo and in vitro. We therefore conclude that the Ku80 carboxy terminus is important to support DNA-PK(CS) autophosphorylation at specific sites, which facilitates DNA end processing by the Artemis...

  13. Identification of a tetramerization domain in the C terminus of the vanilloid receptor.

    Science.gov (United States)

    García-Sanz, Nuria; Fernández-Carvajal, Asia; Morenilla-Palao, Cruz; Planells-Cases, Rosa; Fajardo-Sánchez, Emmanuel; Fernández-Ballester, Gregorio; Ferrer-Montiel, Antonio

    2004-06-09

    TRPV1 (transient receptor potential vanilloid receptor subtype 1) is a member of the TRP channel family gated by vanilloids, protons, and heat. Structurally, TRPV1 appears to be a tetramer formed by the assembly of four identical subunits around a central aqueous pore. The molecular determinants that govern its subunit oligomerization remain elusive. Here, we report the identification of a segment comprising 684Glu-721Arg (referred to as the TRP-like domain) in the C terminus of TRPV1 as an association domain (AD) of the protein. Purified recombinant C terminus of TRPV1 (TRPV1-C) formed discrete and stable multimers in vitro. Yeast two-hybrid and pull-down assays showed that self-association of the TRPV1-C is blocked when segment 684Glu-721Arg is deleted. Biochemical and immunological analysis indicate that removal of the AD from full-length TRPV1 monomers blocks the formation of stable heteromeric assemblies with wild-type TRPV1 subunits. Deletion of the AD in a poreless TRPV1 subunit suppressed its robust dominant-negative phenotype. Together, these findings are consistent with the tenet that the TRP-like domain in TRPV1 is a molecular determinant of the tetramerization of receptor subunits into functional channels. Our observations suggest that the homologous TRP domain in the TRP protein family may function as a general, evolutionary conserved AD involved in subunit multimerization.

  14. The Ku80 carboxy terminus stimulates joining and artemis-mediated processing of DNA ends.

    Science.gov (United States)

    Weterings, Eric; Verkaik, Nicole S; Keijzers, Guido; Florea, Bogdan I; Wang, Shih-Ya; Ortega, Laura G; Uematsu, Naoya; Chen, David J; van Gent, Dik C

    2009-03-01

    Repair of DNA double-strand breaks (DSBs) is predominantly mediated by nonhomologous end joining (NHEJ) in mammalian cells. NHEJ requires binding of the Ku70-Ku80 heterodimer (Ku70/80) to the DNA ends and subsequent recruitment of the DNA-dependent protein kinase catalytic subunit (DNA-PK(CS)) and the XRCC4/ligase IV complex. Activation of the DNA-PK(CS) serine/threonine kinase requires an interaction with Ku70/80 and is essential for NHEJ-mediated DSB repair. In contrast to previous models, we found that the carboxy terminus of Ku80 is not absolutely required for the recruitment and activation of DNA-PK(CS) at DSBs, although cells that harbored a carboxy-terminal deletion in the Ku80 gene were sensitive to ionizing radiation and showed reduced end-joining capacity. More detailed analysis of this repair defect showed that DNA-PK(CS) autophosphorylation at Thr2647 was diminished, while Ser2056 was phosphorylated to normal levels. This resulted in severely reduced levels of Artemis nuclease activity in vivo and in vitro. We therefore conclude that the Ku80 carboxy terminus is important to support DNA-PK(CS) autophosphorylation at specific sites, which facilitates DNA end processing by the Artemis endonuclease and the subsequent joining reaction.

  15. The Ku80 Carboxy Terminus Stimulates Joining and Artemis-Mediated Processing of DNA Ends▿

    Science.gov (United States)

    Weterings, Eric; Verkaik, Nicole S.; Keijzers, Guido; Florea, Bogdan I.; Wang, Shih-Ya; Ortega, Laura G.; Uematsu, Naoya; Chen, David J.; van Gent, Dik C.

    2009-01-01

    Repair of DNA double-strand breaks (DSBs) is predominantly mediated by nonhomologous end joining (NHEJ) in mammalian cells. NHEJ requires binding of the Ku70-Ku80 heterodimer (Ku70/80) to the DNA ends and subsequent recruitment of the DNA-dependent protein kinase catalytic subunit (DNA-PKCS) and the XRCC4/ligase IV complex. Activation of the DNA-PKCS serine/threonine kinase requires an interaction with Ku70/80 and is essential for NHEJ-mediated DSB repair. In contrast to previous models, we found that the carboxy terminus of Ku80 is not absolutely required for the recruitment and activation of DNA-PKCS at DSBs, although cells that harbored a carboxy-terminal deletion in the Ku80 gene were sensitive to ionizing radiation and showed reduced end-joining capacity. More detailed analysis of this repair defect showed that DNA-PKCS autophosphorylation at Thr2647 was diminished, while Ser2056 was phosphorylated to normal levels. This resulted in severely reduced levels of Artemis nuclease activity in vivo and in vitro. We therefore conclude that the Ku80 carboxy terminus is important to support DNA-PKCS autophosphorylation at specific sites, which facilitates DNA end processing by the Artemis endonuclease and the subsequent joining reaction. PMID:19103741

  16. Chemo-spectroscopic sensor for carboxyl terminus overexpressed in carcinoma cell membrane.

    Science.gov (United States)

    Stanca, Sarmiza E; Matthäus, Christian; Neugebauer, Ute; Nietzsche, Sandor; Fritzsche, Wolfgang; Dellith, Jan; Heintzmann, Rainer; Weber, Karina; Deckert, Volker; Krafft, Christoph; Popp, Jürgen

    2015-10-01

    Certain carboxyl groups of the plasma membrane are involved in tumorgenesis processes. A gold core-hydroxyapatite shell (AuHA) nanocomposite is introduced as chemo-spectroscopic sensor to monitor these carboxyl groups of the cell membrane. Hydroxyapatite (HA) plays the role both of a chemical detector and of a biocompatible Raman marker. The principle of detection is based on chemical interaction between the hydroxyl groups of the HA and the carboxyl terminus of the proteins. The AuHA exhibits a surface enhanced Raman scattering (SERS) signal at 954 cm(-1) which can be used for its localization. The bio-sensing capacity of AuHA towards human skin epidermoid carcinoma (A431) and Chinese hamster ovary (CHO) cell lines is investigated using Raman microspectroscopic imaging. The localization of AuHA on cells is correlated with scanning electron microscopy, transmission electron microscopy and structured illumination fluorescence microscopy. This qualitative approach is a step towards a quantitative study of the proteins terminus. This method would enable further studies on the molecular profiling of the plasma membrane, in an attempt to provide accurate cell identification. Using a gold core-hydroxyapatite shell (AuHA) nanocomposite, the authors in this paper showed the feasibility of detecting and differentiating cell surface molecules by surface enhanced Raman scattering. Copyright © 2015 Elsevier Inc. All rights reserved.

  17. Numerical Modeling of Climatic Change from the Terminus Record of Lewis Glacier, Mount Kenya.

    Science.gov (United States)

    Kruss, Phillip Donald

    Over the last 100 years, the glaciers and lakes of East Africa have undergone dramatic change in response to climatic forcing. However, the available conventional meterological series have not proven sufficient to explain these environmental events. The secular climatic change at Lewis Glacier, Mount Kenya (0(DEGREES)9'S, 37(DEGREES)19'E), is reconstructed from its terminus record documented since 1893. The short-time-step numerical model developed for this study consists of climate and ice dynamics segments. The climate segment directly computes the effect on the net balance of change in the four forcings: precipitation, albedo, cloudiness, and temperature. The flow segment calculates the dynamic glacier response to net balance variation. Climatic change occurs over a wide range of time scales. Each glacier responds in a unique fashion to this spectrum of climatic forcings. The response of the Lewis terminus extent to repeated sinusoidal fluctuation in the net balance is calculated. The net balance versus elevation profile is separately translated along the orthogonal balance and elevation axes. Net balance amplitudes of 0.1 to 0.5 m a('-1) of ice and 10 to 50 m elevation, respectively, and periods ranging from 20 to 1000 years are covered. Consideration of the Lewis response is perspective with similar results for Hintereisferner, Storglaciaren, and Berendon and South Cascade Glaciers identifies general characteristics of the time lag and amplitude of the terminus response. The magnitude and timing of the change in only one of the climatic forcings precipitation, albedo, cloudiness, or temperature necessary to produce the retreat of the Lewis terminus from its late 19th century maximum are computed. Equivalent changes for two scenarios of simultaneous variation, namely precipitation/albedo/cloudiness and temperature/albedo, are also estimated. These numerical results are interpreted in the light of long-term lake level, river flow, and instrumental information. A

  18. The N-terminus of glycogen phosphorylase b is not required for activation by adenosine 5'-monophosphate.

    Science.gov (United States)

    Bigley, Andrew N; Reinhart, Gregory D

    2010-06-15

    The, so far unsuccessful, search for selective effective inhibitors of glycogen phosphorylase for the treatment of type II diabetes has made phosphorylase an active target of research for the past 20 years. Many crystallographic structures of phosphorylase are currently available to aid in this research. However, those structures have been interpreted, at least in part, on the basis of work conducted with a proteolytically derived form of phosphorylase that lacked the N-terminus (phosphorylase b'). It has been reported that phosphorylase b' shows no allostery, neither homotropic nor heterotropic. The original report on phosphorylase b' examined the allosteric characteristics over very narrow ranges of effector and substrate concentrations and reported the presence of proteolytic cleavages in addition to the removal of the N-terminus. We have applied molecular biological techniques to generate a truncate lacking the N-terminus with known primary structure, and we have established conditions for fully quantifying the allosteric effect of AMP on glycogen phosphorylase b. We report here for the first time the full thermodynamic effect of AMP on phosphorylase b. Our findings with a truncate lacking the N-terminus show that the effect of AMP binding does not depend on the N-terminus.

  19. Create Your Plate

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    Full Text Available ... Plate Share Create Your Plate ! Share: Seven Simple Steps to Create Your Plate It's simple and effective ... foods within each food category. Try these seven steps to get started: Using your dinner plate, put ...

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  1. Create Your Plate

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    Full Text Available ... of the differences in types of vegetables. When creating your plate at home, remember that half of ... effective for both managing diabetes and losing weight. Creating your plate lets you still choose the foods ...

  2. The Cu(II) affinity of the N-terminus of human copper transporter CTR1: Comparison of human and mouse sequences.

    Science.gov (United States)

    Bossak, Karolina; Drew, Simon C; Stefaniak, Ewelina; Płonka, Dawid; Bonna, Arkadiusz; Bal, Wojciech

    2018-05-01

    Copper Transporter 1 (CTR1) is a homotrimeric membrane protein providing the main route of copper transport into eukaryotic cells from the extracellular milieu. Its N-terminal extracellular domain, rich in His and Met residues, is considered responsible for directing copper into the transmembrane channel. Most of vertebrate CTR1 proteins contain the His residue in position three from N-terminus, creating a well-known Amino Terminal Cu(II)- and Ni(II)-Binding (ATCUN) site. CTR1 from humans, primates and many other species contains the Met-Asp-His (MDH) sequence, while some rodents including mouse have the Met-Asn-His (MNH) N-terminal sequence. CTR1 is thought to collect Cu(II) ions from blood copper transport proteins, including albumin, but previous reports indicated that the affinity of N-terminal peptide/domain of CTR1 is significantly lower than that of albumin, casting serious doubt on this aspect of CTR1 function. Using potentiometry and spectroscopic techniques we demonstrated that MDH-amide, a tripeptide model of human CTR1 N-terminus, binds Cu(II) with K of 1.3 × 10 13  M -1 at pH 7.4, ~13 times stronger than Human Serum Albumin (HSA), and MNH-amide is even stronger, K of 3.2 × 10 14  M -1 at pH 7.4. These results indicate that the N-terminus of CTR1 may serve as intermediate binding site during Cu(II) transfer from blood copper carriers to the transporter. MDH-amide, but not MNH-amide also forms a low abundance complex with non-ATCUN coordination involving the Met amine, His imidazole and Asp carboxylate. This species might assist Cu(II) relay down the peptide chain or its reduction to Cu(I), both steps necessary for the CTR1 function. Copyright © 2018 Elsevier Inc. All rights reserved.

  3. Properties of GluR3 receptors tagged with GFP at the amino or carboxyl terminus.

    Science.gov (United States)

    Limon, Agenor; Reyes-Ruiz, Jorge Mauricio; Eusebi, Fabrizio; Miledi, Ricardo

    2007-09-25

    Anatomical visualization of neurotransmitter receptor localization is facilitated by tagging receptors, but this process can alter their functional properties. We have evaluated the distribution and properties of WT glutamate receptor 3 (GluR3) alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA) receptors (WT GluR3) and two receptors in which GFP was tagged to the amino terminus (GFP-GluR3) or to the carboxyl terminus (GluR3-GFP). Although the fluorescence in Xenopus oocytes was stronger in the vegetal hemisphere because of localization of internal structures (probable sites of production, storage or recycling of receptors), the insertion of receptors into the plasma membrane was polarized to the animal hemisphere. The fluorescence intensity of oocytes injected with GluR3-GFP RNA was approximately double that of oocytes injected with GFP-GluR3 RNA. Accordingly, GluR3-GFP oocytes generated larger kainate-induced currents than GFP-GluR3 oocytes, with similar EC(50) values. Currents elicited by glutamate, or AMPA coapplied with cyclothiazide, were also larger in GluR3-GFP oocytes. The glutamate- to kainate-current amplitude ratios differed, with GluR3-GFP being activated more efficiently by glutamate than the WT or GFP-GluR3 receptors. This pattern correlates with the slower decay of glutamate-induced currents generated by GluR3-GFP receptors. These changes were not observed when GFP was tagged to the amino terminus, and these receptors behaved like the WT. The antagonistic effects of 6-nitro-7-sulfamoylbenzo[f]quinoxaline-2,3-dione (NBQX) and 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) were not altered in any of the tagged receptors. We conclude that GFP is a useful and convenient tag for visualizing these proteins. However, the effects of different sites of tag insertion on receptor characteristics must be taken into account in assessing the roles played by these receptor proteins.

  4. 5,7-DHT lesion of the dorsal raphe nuclei impairs object recognition but not affective behavior and corticosterone response to stressor in the rat.

    Science.gov (United States)

    Lieben, Cindy K J; Steinbusch, Harry W M; Blokland, Arjan

    2006-04-03

    Previous studies with acute tryptophan depletion, leading to transient central 5-HT reductions, showed no effects on affective behavior but impaired object memory. In the present study, the behavioral effects of a 5,7-dihydroxytryptamine (5,7-DHT) lesion in the dorsal raphe were evaluated in animal models of anxiety (open field test), depression (forced swimming test), behavioral inhibition (discrete fixed interval test) and cognition (object recognition task). The corticosterone response to a stress condition was examined at several intervals after 5,7-DHT treatment. The substantial reduction in neuronal 5-HT markers in the dorsal raphe did not affect anxiety-related, depressive-like or impulsive behavior. Compared to the SHAM group, the lesioned rats showed a lower response latency to obtain a reward, indicating a quick and accurate reaction to a stimulus. No differences were found in the progressive ratio test for food motivation. A marked impairment in object recognition was found. The 5,7-DHT treatment did not affect the corticosterone response to a stressful situation. Overall, these results corroborate studies with acute tryptophan depletion suggesting a role of 5-HT in object memory, but not affective behavior.

  5. Differential regulation of serotonin-1A receptor-stimulated [35S]GTP gamma S binding in the dorsal raphe nucleus by citalopram and escitalopram.

    Science.gov (United States)

    Rossi, Dania V; Burke, Teresa F; Hensler, Julie G

    2008-03-31

    The effect of chronic citalopram or escitalopram administration on 5-HT1A receptor function in the dorsal raphe nucleus was determined by measuring [35S]GTP gamma S binding stimulated by the 5-HT1A receptor agonist (R)-(+)-8-OH-DPAT (1nM-10 microM). Although chronic administration of citalopram or escitalopram has been shown to desensitize somatodendritic 5-HT1A autoreceptors, we found that escitalopram treatment decreased the efficacy of 5-HT1A receptors to activate G proteins, whereas citalopram treatment did not. The binding of [3H]8-OH-DPAT to the coupled, high affinity agonist state of the receptor was not altered by either treatment. Interestingly, escitalopram administration resulted in greater occupancy of serotonin transporter sites as measured by the inhibition of [3H]cyanoimipramine binding. As the binding and action of escitalopram is limited by the inactive enantiomer R-citalopram present in racemic citalopram, we propose that the regulation of 5-HT1A receptor function in the dorsal raphe nucleus at the level of receptor-G protein interaction may be a result of greater inhibition of the serotonin transporter by escitalopram.

  6. Differential regulation of serotonin-1A receptor stimulated [35S]GTPγS binding in the dorsal raphe nucleus by citalopram and escitalopram

    Science.gov (United States)

    Rossi, Dania V.; Burke, Teresa F.; Hensler, Julie G.

    2008-01-01

    The effect of chronic citalopram or escitalopram administration on 5-HT1A receptor function in the dorsal raphe nucleus was determined by measuring [35S]GTPγS binding stimulated by the 5-HT1A receptor agonist (R)-(+)-8-OH-DPAT (1nM-10μM). Although chronic administration of citalopram or escitalopram has been shown to desensitize somatodendritic 5-HT1A autoreceptors, we found that escitalopram treatment decreased the efficacy of 5-HT1A receptors to activate G-proteins, whereas citalopram treatment did not. The binding of [3H]8-OH-DPAT to the coupled, high affinity agonist state of the receptor was not altered by either treatment. Interestingly, escitalopram administration resulted in greater occupancy of serotonin transporter sites as measured by the inhibition of [3H]cyanoimipramine binding. As the binding and action of escitalopram is limited by the inactive enantiomer R-citalopram present in racemic citalopram, we propose that the regulation of 5-HT1A receptor function in the dorsal raphe nucleus at the level of receptor-G protein interaction may be a result of greater inhibition of the serotonin transporter by escitalopram. PMID:18289523

  7. Effects of electroacupuncture on orphanin FQ immunoreactivity and preproorphanin FQ mRNA in nucleus of raphe magnus in the neuropathic pain rats.

    Science.gov (United States)

    Ma, Fei; Xie, Hong; Dong, Zhi-Qiang; Wang, Yan-Qing; Wu, Gen-Cheng

    2004-07-15

    Orphanin FQ (OFQ) is an endogenous ligand for opioid receptor-like-1 (ORL1) receptor. Previous studies have shown that both OFQ immunoreactivity and preproorphanin FQ (ppOFQ) mRNA expression could be observed in the brain regions involved in pain modulation, e.g., nucleus of raphe magnus (NRM), dorsal raphe nucleus (DRN), and ventrolateral periaqueductal gray (vlPAG). It was reported that electroacupuncture (EA) has analgesic effect on neuropathic pain, and the analgesic effect was mediated by the endogenous opioid peptides. In the present study, we investigated the effects of EA on the changes of OFQ in the neuropathic pain rats. In the sciatic nerve chronic constriction injury (CCI) model, we investigated the changes of ppOFQ mRNA and OFQ immunoreactivity in NRM after EA by in situ hybridization (ISH) and immunohistochemistry methods, respectively. Then, the ppOFQ mRNA-positive and OFQ immunoreactive cells were counted under a computerized image analysis system. The results showed that expression of ppOFQ mRNA decreased and OFQ immunoreactivity increased after EA treatment in the neuropathic pain rats. These results indicated that EA modulated OFQ synthesis and OFQ peptide level in NRM of the neuropathic pain rats. Copyright 2004 Elsevier Inc.

  8. Thermal Synthesis of Polypeptides from N-Butyloxycarbonyl Oligopeptides Containing Aspartyl Residue at C-Terminus

    Directory of Open Access Journals (Sweden)

    Toratane Munegumi

    2017-01-01

    Full Text Available The thermal reactions of amino acids have been investigated for pure organic synthesis, materials preparation in industry, and prebiotic chemistry. N-t-Butyloxycarbonyl aspartic acid (Boc-Asp releases 2-butene and carbon dioxide upon heating without solvents. The resulting mixture of the free molten aspartic acid was dehydrated to give peptide bonds. This study describes the thermal reactions of N-t-butyloxycarbonyl peptides (Boc-Gly-L-Asp, Boc-L-Ala-L-Asp, Boc-L-Val-L-Asp, and Boc-Gly-Gly-L-Asp having an aspartic residue at the carboxyl terminus. The peptides were deprotected upon heating at a constant temperature between 110 and 170°C for 1 to 24 h to afford polypeptides in which the average molecular weight reached 7800.

  9. C-terminus-mediated voltage gating of Arabidopsis guard cell anion channel QUAC1.

    Science.gov (United States)

    Mumm, Patrick; Imes, Dennis; Martinoia, Enrico; Al-Rasheid, Khaled A S; Geiger, Dietmar; Marten, Irene; Hedrich, Rainer

    2013-09-01

    Anion transporters in plants play a fundamental role in volume regulation and signaling. Currently, two plasma membrane-located anion channel families—SLAC/SLAH and ALMT—are known. Among the ALMT family, the root-expressed ALuminium-activated Malate Transporter 1 was identified by comparison of aluminum-tolerant and Al(3+)-sensitive wheat cultivars and was subsequently shown to mediate voltage-independent malate currents. In contrast, ALMT12/QUAC1 (QUickly activating Anion Channel1) is expressed in guard cells transporting malate in an Al(3+)-insensitive and highly voltage-dependent manner. So far, no information is available about the structure and mechanism of voltage-dependent gating with the QUAC1 channel protein. Here, we analyzed gating of QUAC1-type currents in the plasma membrane of guard cells and QUAC1-expressing oocytes revealing similar voltage dependencies and activation–deactivation kinetics. In the heterologous expression system, QUAC1 was electrophysiologically characterized at increasing extra- and intracellular malate concentrations. Thereby, malate additively stimulated the voltage-dependent QUAC1 activity. In search of structural determinants of the gating process, we could not identify transmembrane domains common for voltage-sensitive channels. However, site-directed mutations and deletions at the C-terminus of QUAC1 resulted in altered voltage-dependent channel activity. Interestingly, the replacement of a single glutamate residue, which is conserved in ALMT channels from different clades, by an alanine disrupted QUAC1 activity. Together with C- and N-terminal tagging, these results indicate that the cytosolic C-terminus is involved in the voltage-dependent gating mechanism of QUAC1.

  10. New bradykinin analogues acetylated on their N-terminus: Effect on rat blood pressure and rat uterus

    Czech Academy of Sciences Publication Activity Database

    Labudda-Dawidowska, O.; Sobolewski, D.; Sleszyňska, M.; Derdowska, I.; Slaninová, Jiřina; Wierzba, T.; Prahl, A.

    2006-01-01

    Roč. 12, Supplement (2006), s. 190 ISSN 1075-2617. [European Peptide Symposium /29./. 03.09.2006-08.09.2006, Gdansk] Institutional research plan: CEZ:AV0Z40550506 Keywords : bradykinin * N-terminus * rat uterus Subject RIV: CC - Organic Chemistry

  11. Unprecedented large inverted repeats at the replication terminus of circular bacterial chromosomes suggest a novel mode of chromosome rescue

    Science.gov (United States)

    El Kafsi, Hela; Loux, Valentin; Mariadassou, Mahendra; Blin, Camille; Chiapello, Hélène; Abraham, Anne-Laure; Maguin, Emmanuelle; van de Guchte, Maarten

    2017-01-01

    The first Lactobacillus delbrueckii ssp. bulgaricus genome sequence revealed the presence of a very large inverted repeat (IR), a DNA sequence arrangement which thus far seemed inconceivable in a non-manipulated circular bacterial chromosome, at the replication terminus. This intriguing observation prompted us to investigate if similar IRs could be found in other bacteria. IRs with sizes varying from 38 to 76 kbp were found at the replication terminus of all 5 L. delbrueckii ssp. bulgaricus chromosomes analysed, but in none of 1373 other chromosomes. They represent the first naturally occurring very large IRs detected in circular bacterial genomes. A comparison of the L. bulgaricus replication terminus regions and the corresponding regions without IR in 5 L. delbrueckii ssp. lactis genomes leads us to propose a model for the formation and evolution of the IRs. The DNA sequence data are consistent with a novel model of chromosome rescue after premature replication termination or irreversible chromosome damage near the replication terminus, involving mechanisms analogous to those proposed in the formation of very large IRs in human cancer cells. We postulate that the L. delbrueckii ssp. bulgaricus-specific IRs in different strains derive from a single ancestral IR of at least 93 kbp. PMID:28281695

  12. Hypocretin/Orexin Peptides Alter Spike Encoding by Serotonergic Dorsal Raphe Neurons through Two Distinct Mechanisms That Increase the Late Afterhyperpolarization.

    Science.gov (United States)

    Ishibashi, Masaru; Gumenchuk, Iryna; Miyazaki, Kenichi; Inoue, Takafumi; Ross, William N; Leonard, Christopher S

    2016-09-28

    Orexins (hypocretins) are neuropeptides that regulate multiple homeostatic processes, including reward and arousal, in part by exciting serotonergic dorsal raphe neurons, the major source of forebrain serotonin. Here, using mouse brain slices, we found that, instead of simply depolarizing these neurons, orexin-A altered the spike encoding process by increasing the postspike afterhyperpolarization (AHP) via two distinct mechanisms. This orexin-enhanced AHP (oeAHP) was mediated by both OX1 and OX2 receptors, required Ca(2+) influx, reversed near EK, and decayed with two components, the faster of which resulted from enhanced SK channel activation, whereas the slower component decayed like a slow AHP (sAHP), but was not blocked by UCL2077, an antagonist of sAHPs in some neurons. Intracellular phospholipase C inhibition (U73122) blocked the entire oeAHP, but neither component was sensitive to PKC inhibition or altered PKA signaling, unlike classical sAHPs. The enhanced SK current did not depend on IP3-mediated Ca(2+) release but resulted from A-current inhibition and the resultant spike broadening, which increased Ca(2+) influx and Ca(2+)-induced-Ca(2+) release, whereas the slower component was insensitive to these factors. Functionally, the oeAHP slowed and stabilized orexin-induced firing compared with firing produced by a virtual orexin conductance lacking the oeAHP. The oeAHP also reduced steady-state firing rate and firing fidelity in response to stimulation, without affecting the initial rate or fidelity. Collectively, these findings reveal a new orexin action in serotonergic raphe neurons and suggest that, when orexin is released during arousal and reward, it enhances the spike encoding of phasic over tonic inputs, such as those related to sensory, motor, and reward events. Orexin peptides are known to excite neurons via slow postsynaptic depolarizations. Here we elucidate a significant new orexin action that increases and prolongs the postspike

  13. Determining the apical terminus of root-end resected teeth using three modern apex locators: a comparative ex vivo study.

    Science.gov (United States)

    ElAyouti, A; Kimionis, I; Chu, A-L; Löst, C

    2005-11-01

    To assess ex vivo the accuracy of various electronic apex locators in locating the apical terminus of root-end resected teeth. Ninety extracted human posterior teeth (182 root canals) were prepared to a minimum size of 40 and filled with gutta-percha and sealer. After resection of the apical 3 mm of the root, the root canal filling was removed using HERO rotary instruments. The size of the root canal at the apical terminus after removal of the filling ranged from size 50 to 90. The root canal length to the apical terminus was determined using 3 apex locators (Root ZX, Raypex4 and Apex Pointer). A new mounting model that utilized a micrometer was used to perform the measurements and to visually determine the actual position of the apical terminus. The frequency of locating the apical terminus and the corresponding 95% confidence interval (CI) were calculated. Additionally, the coefficient of repeatability of each apex locator and the limits of inter-operator agreement were determined. All apex locators showed an acceptable repeatability (0.02-0.03 mm coefficient of repeatability) and narrow limits of inter-operator agreement (+0.07 and -0.07 mm). The accuracy of determining the apical terminus within 1 mm in the root canal was as follows: Root ZX 90% (164/182 root-canals) [95%CI: 86-94%], Raypex4 74% (135/182 root-canals) [95%CI: 68-80%], and Apex Pointer 71% (129/182 root canals) [95%CI: 65-77%]. No over-instrumentation resulted when the Root ZX device was used. In contrast, using the Raypex4 or the Apex Pointer device resulted in over-instrumentation in 8 of 182 root canals (4%). Under the conditions of this study all three apex locators were able to detect the apical terminus of root-end resected teeth with an acceptable range. The Root ZX device was the most accurate without over-instrumentation of the root canals.

  14. Create Your Plate

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    Full Text Available ... In Memory In Honor Become a Member En Español Type 1 Type 2 About Us Online Community ... Page Text Size: A A A Listen En Español Create Your Plate Create Your Plate is a ...

  15. Create Your Plate

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    Full Text Available ... Diabetes Meal Plans Create Your Plate Gluten Free Diets Meal Planning for Vegetarian Diets Cook with Heart-Healthy Foods Holiday Meal Planning ... Planning Meals Diabetes Meal Plans and a Healthy Diet Create Your Plate Meal Planning for Vegetarian Diets ...

  16. Create Your Plate

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    Full Text Available ... Planning Meals Diabetes Meal Plans Create Your Plate Gluten Free Diets Meal Planning for Vegetarian Diets Cook with Heart- ... Create Your Plate Meal Planning for Vegetarian Diets Gluten Free Diets Holiday Meal Planning Cook with Heart-Healthy Foods ...

  17. Create Your Plate

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    Full Text Available ... Planning Meals > Create Your Plate Share: Print Page Text Size: A A A Listen En Español Create ... Type 2 Education Series Hear audio clips and full recordings of past Q&A events at your ...

  18. Amyloid fibril formation of peptides derived from the C-terminus of CETP modulated by lipids

    Energy Technology Data Exchange (ETDEWEB)

    García-González, Victor [Instituto de Fisiología Celular, Universidad Nacional Autónoma de México, 04510 México, DF (Mexico); Mas-Oliva, Jaime, E-mail: jmas@ifc.unam.mx [Instituto de Fisiología Celular, Universidad Nacional Autónoma de México, 04510 México, DF (Mexico); División de Investigación, Facultad de Medicina, Universidad Nacional Autónoma de México, 04510 México, DF (Mexico)

    2013-04-26

    Highlights: •The secondary structure of a C-terminal peptide derived from CETP was studied. •Lipids modulate secondary structure changes of a C-terminal peptide derived from CETP. •Lysophosphatidic acid maintains a functional α-helix and prevents fibril formation. •Transfer of lipids by CETP is related to the presence of an α-helix at its C-end. -- Abstract: Cholesteryl-ester transfer protein (CETP) is a plasmatic protein involved in neutral lipid transfer between lipoproteins. Focusing on the last 12 C-terminus residues we have previously shown that mutation D{sub 470}N promotes a conformational change towards a β-secondary structure. In turn, this modification leads to the formation of oligomers and fibrillar structures, which cause cytotoxic effects similar to the ones provoked by amyloid peptides. In this study, we evaluated the role of specific lipid arrangements on the structure of peptide helix-Z (D{sub 470}N) through the use of thioflavin T fluorescence, peptide bond absorbance, circular dichroism and electron microscopy. The results indicate that the use of micelles formed with lysophosphatidylcholine and lysophosphatidic acid (LPA) under neutral pH induce a conformational transition of peptide helix-Z containing a β-sheet conformation to a native α-helix structure, therefore avoiding the formation of amyloid fibrils. In contrast, incubation with phosphatidic acid does not change the profile for the β-sheet conformation. When the electrostatic charge at the surface of micelles or vesicles is regulated through the use of lipids such as phospholipid and LPA, minimal changes and the presence of β-structures were recorded. Mixtures with a positive net charge diminished the percentage of β-structure and the amount of amyloid fibrils. Our results suggest that the degree of solvation determined by the presence of a free hydroxyl group on lipids such as LPA is a key condition that can modulate the secondary structure and the consequent formation of

  19. Amyloid fibril formation of peptides derived from the C-terminus of CETP modulated by lipids

    International Nuclear Information System (INIS)

    García-González, Victor; Mas-Oliva, Jaime

    2013-01-01

    Highlights: •The secondary structure of a C-terminal peptide derived from CETP was studied. •Lipids modulate secondary structure changes of a C-terminal peptide derived from CETP. •Lysophosphatidic acid maintains a functional α-helix and prevents fibril formation. •Transfer of lipids by CETP is related to the presence of an α-helix at its C-end. -- Abstract: Cholesteryl-ester transfer protein (CETP) is a plasmatic protein involved in neutral lipid transfer between lipoproteins. Focusing on the last 12 C-terminus residues we have previously shown that mutation D 470 N promotes a conformational change towards a β-secondary structure. In turn, this modification leads to the formation of oligomers and fibrillar structures, which cause cytotoxic effects similar to the ones provoked by amyloid peptides. In this study, we evaluated the role of specific lipid arrangements on the structure of peptide helix-Z (D 470 N) through the use of thioflavin T fluorescence, peptide bond absorbance, circular dichroism and electron microscopy. The results indicate that the use of micelles formed with lysophosphatidylcholine and lysophosphatidic acid (LPA) under neutral pH induce a conformational transition of peptide helix-Z containing a β-sheet conformation to a native α-helix structure, therefore avoiding the formation of amyloid fibrils. In contrast, incubation with phosphatidic acid does not change the profile for the β-sheet conformation. When the electrostatic charge at the surface of micelles or vesicles is regulated through the use of lipids such as phospholipid and LPA, minimal changes and the presence of β-structures were recorded. Mixtures with a positive net charge diminished the percentage of β-structure and the amount of amyloid fibrils. Our results suggest that the degree of solvation determined by the presence of a free hydroxyl group on lipids such as LPA is a key condition that can modulate the secondary structure and the consequent formation of amyloid

  20. A Neural Correlate of Predicted and Actual Reward-Value Information in Monkey Pedunculopontine Tegmental and Dorsal Raphe Nucleus during Saccade Tasks

    Science.gov (United States)

    Okada, Ken-ichi; Nakamura, Kae; Kobayashi, Yasushi

    2011-01-01

    Dopamine, acetylcholine, and serotonin, the main modulators of the central nervous system, have been proposed to play important roles in the execution of movement, control of several forms of attentional behavior, and reinforcement learning. While the response pattern of midbrain dopaminergic neurons and its specific role in reinforcement learning have been revealed, the role of the other neuromodulators remains rather elusive. Here, we review our recent studies using extracellular recording from neurons in the pedunculopontine tegmental nucleus, where many cholinergic neurons exist, and the dorsal raphe nucleus, where many serotonergic neurons exist, while monkeys performed eye movement tasks to obtain different reward values. The firing patterns of these neurons are often tonic throughout the task period, while dopaminergic neurons exhibited a phasic activity pattern to the task event. The different modulation patterns, together with the activity of dopaminergic neurons, reveal dynamic information processing between these different neuromodulator systems. PMID:22013541

  1. Excitotoxic median raphe lesions aggravate working memory storage performance deficits caused by scopolamine infusion into the dentate gyrus of the hippocampus in the inhibitory avoidance task in rats

    Directory of Open Access Journals (Sweden)

    Babar E.

    2002-01-01

    Full Text Available The interactions between the median raphe nucleus (MRN serotonergic system and the septohippocampal muscarinic cholinergic system in the modulation of immediate working memory storage performance were investigated. Rats with sham or ibotenic acid lesions of the MRN were bilaterally implanted with cannulae in the dentate gyrus of the hippocampus and tested in a light/dark step-through inhibitory avoidance task in which response latency to enter the dark compartment immediately after the shock served as a measure of immediate working memory storage. MRN lesion per se did not alter response latency. Post-training intrahippocampal scopolamine infusion (2 and 4 µg/side produced a more marked reduction in response latencies in the lesioned animals compared to the sham-lesioned rats. Results suggest that the immediate working memory storage performance is modulated by synergistic interactions between serotonergic projections of the MRN and the muscarinic cholinergic system of the hippocampus.

  2. Creating more effective graphs

    CERN Document Server

    Robbins, Naomi B

    2012-01-01

    A succinct and highly readable guide to creating effective graphs The right graph can be a powerful tool for communicating information, improving a presentation, or conveying your point in print. If your professional endeavors call for you to present data graphically, here's a book that can help you do it more effectively. Creating More Effective Graphs gives you the basic knowledge and techniques required to choose and create appropriate graphs for a broad range of applications. Using real-world examples everyone can relate to, the author draws on her years of experience in gr

  3. Interaction between estradiol and 5-HT1A receptors in the median raphe nucleus on acquisition of aversive information and association to the context in ovariectomized rats

    Directory of Open Access Journals (Sweden)

    Telma Gonçalves Carneiro Spera de Andrade

    2017-12-01

    Full Text Available The median raphe nucleus (MRN is related to stress resistance and defensive responses, a crucial source of serotonergic neurons that project to prosencephalic structures related to stress and anxiety. Estrogen receptors were identified in this mesencephalic structure. It is possible that the estrogen action is related to serotonin effect on somatodendritic 5-HT1A receptors, inhibiting the function of serotonergic neurons and thus preventing of the stress effect and inducing anxiolysis. So, in order to evaluate these aspects, female Wistar rats were ovariectomized and 21 days later were given a direct microinjection of estradiol benzoate (EB (1200 ng into the MRN, preceded by microinjections of saline or WAY100.635 (100 ng, a 5-HT1A receptor antagonist. Immediately after the two microinjections, the ovariectomized rats were conditioned with an aversive event (foot shock session in a Skinner box. Twenty-four hours later, they were exposed to the same context in a test session for 5 min for behavioral assessment: freezing, rearing, locomotion, grooming, and autonomic responses (fecal boluses and micturition. EB microinjection in the MRN prior to the exposure of animals to the foot shocks in the conditioning session did not alter their behavior in this session, but neutralized the association of the aversive experience to the context: there was a decrease in the expression of freezing and an increased rearing activity in the test session. This effect was reversed by prior microinjection of WAY100.635. In conclusion, EB acted on serotonergic neurons in the MRN of the ovariectomized rats, impairing the association of the aversive experience to the context, by co-modulating the functionality of somatodendritic 5-HT1A. Keywords: Contextual conditioning, Median raphe nucleus, Estradiol benzoate, 5-HT1A receptors, WAY100.635, Ovariectomized rats, Anxiety

  4. Agonist-dependent modulation of G-protein coupling and transduction of 5-HT1A receptors in rat dorsal raphe nucleus.

    Science.gov (United States)

    Valdizán, Elsa Maria; Castro, Elena; Pazos, Angel

    2010-08-01

    5-HT1A receptors couple to different Go/Gi proteins in order to mediate a wide range of physiological actions. While activation of post-synaptic 5-HT1A receptors is mainly related to inhibition of adenylyl cyclase activity, functionality of autoreceptors located in raphe nuclei has been classically ascribed to modifications of the activity of potassium and calcium channels. In order to evaluate the possible existence of agonist-directed trafficking for 5-HT1A autoreceptors in the rat dorsal raphe nucleus, we studied their activation by two agonists with a different profile of efficacy [(+)8-OH-DPAT and buspirone], addressing simultaneously the identification of the specific Galpha subtypes ([35S]GTPgammaS labelling and immunoprecipitation) involved and the subsequent changes in cAMP formation. A significant increase (32%, plabelling of immunoprecipitates was obtained with anti-Galphai3 antibodies but not with anti-Galphao, anti-Galphai1, anti-Galphai2, anti-Galphaz or anti-Galphas antibodies. In contrast, in the presence of buspirone, significant [35S]GTPgammaS labelling of immunoprecipitates was obtained with anti-Galphai3 (50%, plabelling with anti-Galphai1, anti-Galphaz or anti-Galphas. The selective 5-HT1A antagonist WAY 100635 blocked the labelling induced by both agonists. Furthermore, (+)8-OH-DPAT failed to modify forskolin-stimulated cAMP accumulation, while buspirone induced a dose-dependent, WAY 100635-sensitive, inhibition of this response (Imax 30.8+/-4.9, pIC50 5.95+/-0.46). These results demonstrate the existence of an agonist-dependency pattern of G-protein coupling and transduction for 5-HT1A autoreceptors in native brain tissue. These data also open new perspectives for the understanding of the differential profiles of agonist efficacy in pre- vs. post-synaptic 5-HT1A receptor-associated responses.

  5. Cy3 and Cy5 dyes attached to oligonucleotide terminus stabilize DNA duplexes: predictive thermodynamic model.

    Science.gov (United States)

    Moreira, Bernardo G; You, Yong; Owczarzy, Richard

    2015-03-01

    Cyanine dyes are important chemical modifications of oligonucleotides exhibiting intensive and stable fluorescence at visible light wavelengths. When Cy3 or Cy5 dye is attached to 5' end of a DNA duplex, the dye stacks on the terminal base pair and stabilizes the duplex. Using optical melting experiments, we have determined thermodynamic parameters that can predict the effects of the dyes on duplex stability quantitatively (ΔG°, Tm). Both Cy dyes enhance duplex formation by 1.2 kcal/mol on average, however, this Gibbs energy contribution is sequence-dependent. If the Cy5 is attached to a pyrimidine nucleotide of pyrimidine-purine base pair, the stabilization is larger compared to the attachment to a purine nucleotide. This is likely due to increased stacking interactions of the dye to the purine of the complementary strand. Dangling (unpaired) nucleotides at duplex terminus are also known to enhance duplex stability. Stabilization originated from the Cy dyes is significantly larger than the stabilization due to the presence of dangling nucleotides. If both the dangling base and Cy3 are present, their thermodynamic contributions are approximately additive. New thermodynamic parameters improve predictions of duplex folding, which will help design oligonucleotide sequences for biophysical, biological, engineering, and nanotechnology applications. Copyright © 2015. Published by Elsevier B.V.

  6. The N-terminus of Bunyamwera orthobunyavirus NSs protein is essential for interferon antagonism.

    Science.gov (United States)

    van Knippenberg, Ingeborg; Carlton-Smith, Charlie; Elliott, Richard M

    2010-08-01

    Bunyamwera virus NSs protein is involved in the inhibition of cellular transcription and the interferon (IFN) response, and it interacts with the Med8 component of Mediator. A spontaneous mutant of a recombinant NSs-deleted Bunyamwera virus (rBUNdelNSs2) was identified and characterized. This mutant virus, termed mBUNNSs22, expresses a 21 aa N-terminally truncated form of NSs. Like rBUNdelNSs2, mBUNNSs22 is attenuated in IFN-deficient cells, and to a greater extent in IFN-competent cells. Both rBUNdelNSs2 and mBUNNSs22 are potent IFN inducers and their growth can be rescued by depleting cellular IRF3. Strikingly, despite encoding an NSs protein that contains the Med8 interaction domain, mBUNNSs22 fails to block RNA polymerase II activity during infection. Overall, our data suggest that both the interaction of NSs with Med8 and a novel unidentified function of the NSs N-terminus, seem necessary for Bunyamwera virus to counteract host antiviral responses.

  7. Hpa1 harpin needs nitroxyl terminus to promote vegetative growth and leaf photosynthesis in Arabidopsis.

    Science.gov (United States)

    Li, Xiaojie; Han, Liping; Zhao, Yanying; You, Zhenzhen; Dong, Hansong; Zhang, Chunling

    2014-03-01

    Hpa1 is a harpin protein produced by Xanthomonas oryzae, an important bacterial pathogen of rice, and has the growth-promoting activity in plants. To understand the molecular basis for the function of Hpa1, we generated an inactive variant protein, Hpa1 delta NT, by deleting the nitroxyl-terminal region of the Hpa1 sequence and compared Hpa1 delta NT with the full-length protein in terms of the effects on vegetative growth and related physiological responses in Arabidopsis. When Hpa1 was applied to plants, it acted to enhance the vegetative growth but did not affect the floral development. Enhanced plant growth was accompanied by induced expression of growth-promoting genes in plant leaves. The growth-promoting activity of Hpa1 was further correlated with a physiological consequence shown as promoted leaf photosynthesis as a result of facilitated CO2 conduction through leaf stomata and mesophyll cells. On the contrary, plant growth, growth-promoting gene expression, and the physiological consequence changed little in response to the Hpa1 delta NT treatment. These analyses suggest that Hpa1 requires the nitroxyl-terminus to facilitate CO2 transport inside leaf cells and promote leaf photosynthesis and vegetative growth of the plant.

  8. Truncated C-terminus of fibrillin-1 induces Marfanoid-progeroid-lipodystrophy (MPL) syndrome in rabbit.

    Science.gov (United States)

    Chen, Mao; Yao, Bing; Yang, Qiangbing; Deng, Jichao; Song, Yuning; Sui, Tingting; Zhou, Lina; Yao, HaoBing; Xu, Yuanyuan; Ouyang, Hongsheng; Pang, Daxin; Li, Zhanjun; Lai, Liangxue

    2018-04-09

    Various clinical differences have been observed between patients with the FBN1 gene mutation and those with the classical Marfan phenotype. Although FBN1 knockout (KO) or dominant-negative mutant mice are widely used as an animal model for Marfan syndrome (MFS), these mice cannot recapitulate the genotype/phenotype relationship of Marfanoid-progeroid-lipodystrophy (MPL) syndrome, which is caused by a mutation in the C-terminus of fibrillin-1, the penultimate exon of the FBN1 gene. Here, we describe the generation of a rabbit MPL model with C-terminal truncation of fibrillin-1 using a CRISPR/Cas9 system. FBN1 heterozygous ( FBN1 Het) rabbits faithfully recapitulated the phenotypes of MFS, including muscle wasting and impaired connective tissue, ocular syndrome and aortic dilation. Moreover, skin symptoms, lipodystrophy, growth retardation and dysglycemia were also seen in these FBN1 Het rabbits, and have not been reported in other animal models. In conclusion, this novel rabbit model mimics the histopathological changes and functional defects of MPL syndrome, and could become a valuable model for studies of pathogenesis and drug screening for MPL syndrome. © 2018. Published by The Company of Biologists Ltd.

  9. Oxidative stress induces nuclear translocation of C-terminus of α-synuclein in dopaminergic cells

    International Nuclear Information System (INIS)

    Xu Shengli; Zhou Ming; Yu Shun; Cai Yanning; Zhang Alex; Ueda, Kenji; Chan Piu

    2006-01-01

    Growing evidence suggests that oxidative stress is involved in the neuronal degeneration and can promote the aggregation of α-synuclein. However, the role of α-synuclein under physiological and pathological conditions remains poorly understood. In the present study, we examined the possible interaction between the α-synuclein and oxidative stress. In a dopaminergic cell line MES23.5, we have found that the 200 μM H 2 O 2 treatment induced the translocation of α-synuclein from cytoplasm to nuclei at 30 min post-treatment. The immunoactivity of α-synuclein became highly intensive in the nuclei after 2 h treatment. The protein translocated to nucleus was a 10 kDa fragment of C-terminus region of α-synuclein, while full-length α-synuclein remained in cytoplasm. Thioflavine-S staining suggested that the C-terminal fragment in the nuclei has no β-sheet structures. Our present results indicated that 200 μM H 2 O 2 treatment induces the intranuclear accumulation of the C-terminal fragment of α-synuclein in dopaminergic neurons, whose role remains to be investigated

  10. Create Your Plate

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    Full Text Available ... Food Planning Meals Diabetes Meal Plans and a Healthy Diet Create Your Plate Meal Planning for Vegetarian Diets Gluten Free Diets Holiday Meal Planning Cook with Heart-Healthy Foods donate en -- A Future Without Diabetes - a- ...

  11. Create Your Plate

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    Full Text Available ... Risk Test Lower Your Risk Healthy Eating Overweight Smoking High Blood Pressure Physical Activity High Blood Glucose ... Diabetes Meal Plans Create Your Plate Gluten Free Diets Meal Planning for Vegetarian Diets Cook with Heart- ...

  12. Create Your Plate

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    Full Text Available ... Risk Test Lower Your Risk Healthy Eating Overweight Smoking High Blood Pressure Physical Activity High Blood Glucose ... Planning Meals Diabetes Meal Plans and a Healthy Diet Create Your Plate Meal Planning for Vegetarian Diets ...

  13. Create Your Plate

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    Full Text Available ... Recipes Association Cookbook Recipes Planning Meals Diabetes Meal Plans Create Your Plate Gluten Free Diets Meal Planning ... serving of dairy or both as your meal plan allows. Choose healthy fats in small amounts. For ...

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    Full Text Available ... Food MyFoodAdvisor Recipes Association Cookbook Recipes Planning Meals Diabetes Meal Plans Create Your Plate Gluten Free Diets Meal Planning for Vegetarian Diets Cook with Heart-Healthy Foods Holiday Meal ...

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    Full Text Available ... Index Low-Calorie Sweeteners Sugar and Desserts Fitness Exercise & Type 1 Diabetes Get Started Safely Get And ... Plate Create Your Plate is a simple and effective way to manage your blood glucose levels and ...

  16. Create Your Plate

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    Full Text Available ... Us in the Fight for a Cure Your tax-deductible gift today can fund critical diabetes research ... Close www.diabetes.org > Food and Fitness > Food > Planning Meals > Create Your Plate Share: Print Page Text ...

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    Full Text Available ... critical diabetes research and support vital diabetes education services that improve the lives of those with diabetes. $50 $100 $250 $500 Other Other Ways ... Meals > Create Your Plate ...

  18. Create Your Plate

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  19. Create Your Plate

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    Full Text Available ... Your Plate Gluten Free Diets Meal Planning for Vegetarian Diets Cook with Heart-Healthy Foods Holiday Meal ... Healthy Diet Create Your Plate Meal Planning for Vegetarian Diets Gluten Free Diets Holiday Meal Planning Cook ...

  20. Der terminus προνοητησ in der Byzantinischen verwaltung

    Directory of Open Access Journals (Sweden)

    Wassiliou-Seibt Alexandra-Kyriaki

    2013-01-01

    Full Text Available The article examines on the basis of the small number of essential literary and sigillographic evidence the scope and duties of the προνοηταí (as a terminus technicus in the Byzantine administration. The earliest evidence comes from the 6th century, the latest from the 12th. Already in the Early Byzantine period the scope of their activities is fixed: administration of domains and municipal and fiscal administration. Acting in the capacity of stewards of private and official real estate these people were responsible for collecting and transmitting taxes to the responsible department, minus an allowance for themselves. Their colleagues in the municipal and provincial administration were revenue officers with special authority. In the provincial administration of the 11th the pronoetai were sometimes also άναγραφείς in their region. Till now we know only one example of a commander of a thema who was at the same time also a προνοητής (Eustathios Charsianites. The προνοηταί τού δημοσίου (scil. fiscus were authorized to exact outstanding taxes. A special case is the προνοητής of the alms-house of Michael Attaleiates in Rhaidestos, because he was the highest administrator of all its estates and dependencies. Normally the προνοηταί were administrators of a lower rank in the domains, subject to the οίκονόμος or the κουράτωρ.

  1. The NH2 terminus regulates voltage-dependent gating of CALHM ion channels.

    Science.gov (United States)

    Tanis, Jessica E; Ma, Zhongming; Foskett, J Kevin

    2017-08-01

    Calcium homeostasis modulator protein-1 (CALHM1) and its Caenorhabditis elegans (ce) homolog, CLHM-1, belong to a new family of physiologically important ion channels that are regulated by voltage and extracellular Ca 2+ (Ca 2+ o ) but lack a canonical voltage-sensing domain. Consequently, the intrinsic voltage-dependent gating mechanisms for CALHM channels are unknown. Here, we performed voltage-clamp experiments on ceCLHM-1 chimeric, deletion, insertion, and point mutants to assess the role of the NH 2 terminus (NT) in CALHM channel gating. Analyses of chimeric channels in which the ceCLHM-1 and human (h)CALHM1 NH 2 termini were interchanged showed that the hCALHM1 NT destabilized channel-closed states, whereas the ceCLHM-1 NT had a stabilizing effect. In the absence of Ca 2+ o , deletion of up to eight amino acids from the ceCLHM-1 NT caused a hyperpolarizing shift in the conductance-voltage relationship with little effect on voltage-dependent slope. However, deletion of nine or more amino acids decreased voltage dependence and induced a residual conductance at hyperpolarized voltages. Insertion of amino acids into the NH 2 -terminal helix also decreased voltage dependence but did not prevent channel closure. Mutation of ceCLHM-1 valine 9 and glutamine 13 altered half-maximal activation and voltage dependence, respectively, in 0 Ca 2+ In 2 mM Ca 2+ o , ceCLHM-1 NH 2 -terminal deletion and point mutant channels closed completely at hyperpolarized voltages with apparent affinity for Ca 2+ o indistinguishable from wild-type ceCLHM-1, although the ceCLHM-1 valine 9 mutant exhibited an altered conductance-voltage relationship and kinetics. We conclude that the NT plays critical roles modulating voltage dependence and stabilizing the closed states of CALHM channels. Copyright © 2017 the American Physiological Society.

  2. Creating organizational cultures

    DEFF Research Database (Denmark)

    Mouton, Nicolaas T.O.; Just, Sine Nørholm; Gabrielsen, Jonas

    2012-01-01

    Purpose – The purpose of this paper is to re-conceptualize the relations between rhetorical strategies and material practices in the processes whereby leaders create or change organizational cultures. Design/methodology/approach – The authors compare and contrast two broad perspectives on cultural...... insights. The authors propose an integrated perspective in which material practices and rhetorical strategies are seen as two analytical sides of the same ontological coin. This enables a fuller and more detailed explanation of how organizational cultures are created or changed. A brief illustration...

  3. Creating Web Pages Simplified

    CERN Document Server

    Wooldridge, Mike

    2011-01-01

    The easiest way to learn how to create a Web page for your family or organization Do you want to share photos and family lore with relatives far away? Have you been put in charge of communication for your neighborhood group or nonprofit organization? A Web page is the way to get the word out, and Creating Web Pages Simplified offers an easy, visual way to learn how to build one. Full-color illustrations and concise instructions take you through all phases of Web publishing, from laying out and formatting text to enlivening pages with graphics and animation. This easy-to-follow visual guide sho

  4. Bovine adenovirus type 3 containing heterologous protein in the C-terminus of minor capsid protein IX

    International Nuclear Information System (INIS)

    Zakhartchouk, Alexander; Connors, Wayne; Van Kessel, Andrew; Tikoo, Suresh Kumar

    2004-01-01

    Earlier, we detected pIX of BAdV-3 as a 14-kDa protein in purified virions. Analysis of BAdV-3 pIX using different region antibodies revealed that the N-terminus and central domain of the pIX contain immunogenic sites and are not exposed on the surface of BAdV-3 virion. This suggested that the C-terminus of BAdV-3 pIX (125 amino acid) may be exposed on the virion and may be used as a site for incorporation of heterologous peptides or proteins. We constructed recombinant BAV950 containing a small peptide (21 amino acid), including the RGD motif or recombinant BAV951 containing enhanced yellow-green fluorescent protein (EYFP) fused to the C-terminus of pIX. Western blot analysis demonstrated that the chimeric pIX-RGD was incorporated into virion capsids. Incorporation of the RGD motif into the pIX resulted in significant augmentation of BAdV-3 fiber knob-independent infection of the integrin-positive cells, suggesting that RGD motifs are displayed on the surface of virion capsids and are accessible for binding to integrins. Analysis of BAV951 revealed that the chimeric pIX is incorporated into virion capsids and EYFP containing the C-terminus of pIX is exposed on the surface of the virion. Moreover, insertion of chimeric pIXs was maintained without change through successive rounds of viral replication. These results suggested that in contrast to major capsid proteins (hexon, penton, fiber), the minor capsid protein IX can be use for the incorporation of targeting ligands based on either small peptides or longer polypeptides

  5. Distinct regions in the C-Terminus required for GLP-1R cell surface expression, activity and internalisation.

    Science.gov (United States)

    Thompson, Aiysha; Kanamarlapudi, Venkateswarlu

    2015-09-15

    The glucagon-like peptide-1 (GLP-1) receptor (GLP-1R), an important drug target in the treatment of type 2 diabetes, is a G-protein coupled receptor (GPCR) that mediates insulin secretion by GLP-1. The N-terminus controls GLP-1R biosynthetic trafficking to the cell surface but the C-terminus involvement in that trafficking is unknown. The aim of this study was to identify distinct regions within the C-terminal domain required for human GLP-1R (hGLP-1R) cell surface expression, activity and internalisation using a number of C-terminal deletions and site-directed mutations. The results of this study revealed that the residues 411-418 within the C-terminal domain of the hGLP-1R are critical in targeting the newly synthesised receptor to the plasma membrane. The residues 419-430 are important for cAMP producing activity of the receptor, most likely by coupling to Gαs. However, the residues 431-450 within the C-terminus are essential for agonist-induced hGLP-1R internalisation. In conclusion, these findings demonstrate the hGLP-1R has distinct regions within the C-terminal domain required for its cell surface expression, activity and agonist-induced internalisation. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

  6. Create Your Plate

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    Full Text Available ... meal-planning, . In this section Food Planning Meals Diabetes Meal Plans and a Healthy Diet Create Your Plate Meal Planning for Vegetarian Diets Gluten Free Diets Holiday Meal Planning Cook with Heart-Healthy Foods donate en -- A Future Without Diabetes - a-future-without-diabetes-2.html A Future ...

  7. Creating Customer Delight.

    Science.gov (United States)

    Black, Jim

    1995-01-01

    This article proposes that college admissions officers interested in improving service should focus on creating customer delight rather than simply satisfaction, studying the system when things go wrong rather than placing blame, establishing employee well-being as the highest priority of the organization, providing necessary tools and training…

  8. Create Your Plate

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    Full Text Available ... tool is not to scale because of the differences in types of vegetables. When creating your plate ... function (data) { $('#survey-errors').remove(); $('.survey-form .form-group .survey-alert-wrap').remove(); if (data.submitSurveyResponse.success == ' ...

  9. Creating Pupils' Internet Magazine

    Science.gov (United States)

    Bognar, Branko; Šimic, Vesna

    2014-01-01

    This article presents an action research, which aimed to improve pupils' literary creativity and enable them to use computers connected to the internet. The study was conducted in a small district village school in Croatia. Creating a pupils' internet magazine appeared to be an excellent way for achieving the educational aims of almost all…

  10. Creating a Classroom Makerspace

    Science.gov (United States)

    Rivas, Luz

    2014-01-01

    What is a makerspace? Makerspaces are community-operated physical spaces where people (makers) create do-it-yourself projects together. These membership spaces serve as community labs where people learn together and collaborate on projects. Makerspaces often have tools and equipment like 3-D printers, laser cutters, and soldering irons.…

  11. Create Your Plate

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    Full Text Available ... 1 Type 2 About Us Online Community Meal Planning Sign In Search: Search More Sites Search ≡ Are ... Fitness Home Food MyFoodAdvisor Recipes Association Cookbook Recipes Planning Meals Diabetes Meal Plans Create Your Plate Gluten ...

  12. Creating White Australia

    DEFF Research Database (Denmark)

    McLisky, Claire Louise; Carey, Jane

    Vedtagelsen af White Australien som regeringens politik i 1901 viser, at hvidheden var afgørende for den måde, hvorpå den nye nation i Australien blev konstitueret. Og alligevel har historikere i vid udstrækning overset hvidhed i deres studier af Australiens race fortid. 'Creating White Australia...

  13. Creating Innovative Opportunities

    DEFF Research Database (Denmark)

    Ljungberg, Daniel; McKelvey, Maureen; Lassen, Astrid Heidemann

    2012-01-01

    This paper develops lessons about how and why the founders and ventures involved in knowledge intensive entrepreneurship (KIE) manage the process of venture creation. The meta-analysis of the 86 case studies is based upon as conceptual model (from a systemic literature review), linked to illustra...... of knowledge networks to create innovative opportunities....

  14. Create Your State

    Science.gov (United States)

    Dunham, Kris; Melvin, Samantha

    2011-01-01

    Students are often encouraged to work together with their classmates, sometimes with other classes, occasionally with kids at other schools, but rarely with kids across the country. In this article the authors describe the Create Your State project, a collaborative nationwide project inspired by the Texas Chair Project wherein the artist, Damien…

  15. Creating a Third Culture

    Science.gov (United States)

    Weisbuch, Robert A.

    2008-01-01

    In this article, the author laments higher education's lack of concern towards the development of teaching in the public schools over the last half of the 20th century. Most of academe's work on the topic of teacher training has been done at the branches of state universities that needed to make money and create a niche. The author observes that…

  16. Wann ist ein terminus technicus ein terminus technicus? – Das Beispiel àτρεμής im Corpus Hippocraticum

    Directory of Open Access Journals (Sweden)

    Eva Wöckener-Gade

    2017-07-01

    Full Text Available In this paper, I present some results of my research done in the project eXChange. My main goal was to investigate to what extent and how the terminology of the Corpus Hippocraticum has been influenced by the language of the preceding (esp. lyric literature. A common view in the linguistics concerned with languages for special purposes (the German ‘Fachsprachen’ holds that termini technici are often created by implementing a word from the common language into a technical context and narrowing its former meaning down to a specialized and strictly defined one. While this mechanism could be traced for several terms in the Corpus Hippocraticum, some terms could be found which denote special processes or methods and have been taken over into the medical context without a significant change of meaning. The question is raised if these terms can be regarded as termini technici even if their meaning has not been coined in a specialized context.

  17. Creating resilient SMEs

    DEFF Research Database (Denmark)

    Dahlberg, Rasmus; Guay, Fanny

    2015-01-01

    According to the EU, during the past five years, small and medium enterprises (SMEs) have created 85% of new jobs and two-thirds of private sector employment in the region. SMEs are considered the backbone of the economy in Europe and represent more than 95% of enterprises in USA and Australia....... They are considered more vulnerable to disasters because of their size. This paper argues, on the contrary, that SMEs also can be less vulnerable to sudden change than large corporations, drawing upon the ideas of Hayek and Taleb, and that networks of SMEs may contribute to the overall resilience of society...... if certain criteria are met. With this in mind, this paper will be examining how to create resilient SMEs. A well-known concept in the field is business continuity management. BCM is defined as “a holistic management process that identifies potential threats to an organization and the impacts to business...

  18. Creating flat design websites

    CERN Document Server

    Pratas, Antonio

    2014-01-01

    This book contains practical, step-by-step tutorials along with plenty of explanation about designing your flat website. Each section is introduced sequentially, building up your web design skills and completing your website.Creating Flat Design Websites is ideal for you if you are starting on your web development journey, but this book will also benefit seasoned developers wanting to start developing in flat.

  19. The N terminus of cGAS de-oligomerizes the cGAS:DNA complex and lifts the DNA size restriction of core-cGAS activity.

    Science.gov (United States)

    Lee, Arum; Park, Eun-Byeol; Lee, Janghyun; Choi, Byong-Seok; Kang, Suk-Jo

    2017-03-01

    Cyclic GMP-AMP synthase (cGAS) is a DNA-sensing enzyme in the innate immune system. Recent studies using core-cGAS lacking the N terminus investigated the mechanism for binding of double-stranded (ds) DNA and synthesis of 2',3'-cyclic GMP-AMP (cGAMP), a secondary messenger that ultimately induces type I interferons. However, the function of the N terminus of cGAS remains largely unknown. Here, we found that the N terminus enhanced the activity of core-cGAS in vivo. Importantly, the catalytic activity of core-cGAS decreased as the length of double-stranded DNA (dsDNA) increased, but the diminished activity was restored by addition of the N terminus. Furthermore, the N terminus de-oligomerized the 2 : 2 complex of core-cGAS and dsDNA into a 1 : 1 complex, suggesting that the N terminus enhanced the activity of core-cGAS by facilitating formation of a monomeric complex of cGAS and DNA. © 2017 Federation of European Biochemical Societies.

  20. Creating a practice website.

    Science.gov (United States)

    Downes, P K

    2007-05-26

    A website is a window to the outside world. For a dental practice, it may be the first point of contact for a prospective new patient and will therefore provide them with their 'first impression'; this may be days or weeks before actually visiting the practice. This section considers the different ways of creating a dental practice website and lists some of the main dental website design companies. It also describes what factors make a successful website and offers advice on how to ensure that it complies with current regulations and recommendations.

  1. A morphometric, immunohistochemical, and in situ hybridization study of the dorsal raphe nucleus in major depression, bipolar disorder, schizophrenia, and suicide.

    Science.gov (United States)

    Matthews, Paul R; Harrison, Paul J

    2012-03-01

    Several lines of evidence implicate 5-hydroxytryptamine (5-HT, serotonin) in the pathophysiology of mood disorders and suicide. However, it is unclear whether these conditions include morphological involvement of the dorsal raphe nucleus (DRN), the origin of most forebrain 5-HT innervation. We used morphometric, immunohistochemical, and molecular methods to compare the DRN in post-mortem tissue of 50 subjects (13 controls, 14 major depressive disorder [MDD], 13 bipolar disorder, 10 schizophrenia; 17 of the cases died by suicide). NeuN and PH8 antibodies were used to assess all neurons and serotonergic neurons respectively; 5-HT(1A) autoreceptor expression was investigated by regional and cellular in situ hybridization. Measurements were made at three rostrocaudal levels of the DRN. In MDD, the area of the DRN was decreased. In bipolar disorder, serotonergic neuronal size was decreased. Suicide was associated with an increased DRN area, and with a higher density but decreased size of serotonergic neurons. Total neuronal density and 5-HT(1A) receptor mRNA abundance were unaffected by diagnosis or suicide. No changes were seen in schizophrenia. The results show that mood disorders and suicide are associated with differential, limited morphological alterations of the DRN. The contrasting influences of MDD and suicide may explain some of the discrepancies between previous studies, since their design precluded detection of the effect. Copyright © 2011 Elsevier B.V. All rights reserved.

  2. How does early maternal separation and chronic stress in adult rats affect the immunoreactivity of serotonergic neurons within the dorsal raphe nucleus?

    Science.gov (United States)

    Pollano, Antonella; Trujillo, Verónica; Suárez, Marta M

    2018-01-01

    Vulnerability to emotional disorders like depression derives from interactions between early and late environments, including stressful conditions. The serotonin (5HT) system is strongly affected by stress and chronic unpredictable stress can alter the 5HT system. We evaluated the distribution of active serotonergic neurons in the dorsal raphe nucleus (DR) through immunohistochemistry in maternally separated and chronically stressed rats treated with an antidepressant, tianeptine, whose mechanism of action is still under review. Male Wistar rats were subjected to daily maternal separation (MS) for 4.5 h between postnatal days (PND) 1-21, or to animal facility rearing (AFR). Between (PND) days 50-74, rats were exposed to chronic unpredictable stress and were treated daily with tianeptine (10 mg/kg) or vehicle. We found an interaction between the effects of MS and chronic unpredictable stress on Fos-5HT immunoreactive cells at mid-caudal level of the DR. MS-chronically stressed rats showed an increase of Fos-5HT immunoreactive cells compared with AFR-chronically stressed rats. The ventrolateral (DRL/VLPAG) and dorsal (DRD) subdivisions of the DR were significantly more active than the ventral part (DRV). At the rostral level of the DR, tianeptine decreased the number of Fos-5HT cells in DR in the AFR groups, both unstressed and stressed. Overall, our results support the idea of a match in phenotype exhibited when the early and the adult environment correspond.

  3. Effect of morphine-induced antinociception is altered by AF64A-induced lesions on cholinergic neurons in rat nucleus raphe magnus.

    Science.gov (United States)

    Abe, Kenji; Ishida, Kota; Kato, Masatoshi; Shigenaga, Toshiro; Taguchi, Kyoji; Miyatake, Tadashi

    2002-11-01

    To examine the role of cholinergic neurons in the nucleus raphe magnus (NRM) in noxious heat stimulation and in the effects of morphine-induced antinociception by rats. After the cholinergic neuron selective toxin, AF64A, was microinjected into the NRM, we examined changes in the antinociceptive threshold and effects of morphine (5 mg/kg, ip) using the hot-plate (HP) and tail-flick (TF) tests. Systemic administration of morphine inhibited HP and TF responses in control rats. Microinjection of AF64A (2 nmol/site) into the NRM significantly decreased the threshold of HP response after 14 d, whereas the TF response was not affected. Morphine-induced antinociception was significantly attenuated in rats administered AF64A. Extracellular acetylcholine was attenuated after 14 d to below detectable levels in rats given AF64A. Naloxone (1 microg/site) microinjected into control rat NRM also antagonized the antinociceptive effect of systemic morphine. These findings suggest that cholinergic neuron activation in the NRM modulates the antinociceptive effect of morphine simultaneously with the opiate system.

  4. [The relationships among raphe magnus nucleus, locus coeruleus and dorsal motor nucleus of vagus in the descending regulation of gastric motility].

    Science.gov (United States)

    Qiao, Hui; An, Shu-Cheng; Xu, Chang

    2011-02-01

    To explore the interrelationship among dorsal motor nucleus of the vagus (DMV), locus coeruleus (LC) and raphe magnus nucleus (NRM) in the mechanism of the descending regulation on gastric motility, which may constitute a parasympathetic local circuit, work as a neural center of gastric modulation in brainstem. Using nucleus location, electric stimulation and lesion, together with microinjection, and recording the inter-gastric pressure. (1) LC stimulation could inhibit the gastric motility significantly (P effect, while blocking the a receptor on DMV could reverse the effect. (2) NRM stimulation reduced the amplitude of gastric constriction (P effect, but blocking the 5-HT2A receptor on DMV depressed the gastric motility heavily (P effect of NRM stimulation, and microinjection of ritanserin into LC could likewise abolish it. (1) LC inhibit the gastric motility via a receptor in DMV, and meanwhile may excite it through 5-HT2A receptor in DMV, these two ways work together to keeping the gastric motility amplitude normally. (2) NRM inhibit the gastric motility via 5-HT2A receptor in LC.

  5. Anti-nociceptive effects of calcitonin gene-related peptide in nucleus raphe magnus of rats: an effect attenuated by naloxone.

    Science.gov (United States)

    Huang, Y; Brodda-Jansen, G; Lundeberg, T; Yu, L C

    2000-08-04

    The present study investigated the role of calcitonin gene-related peptide (CGRP) on nociception in nucleus raphe magnus (NRM) and the interaction between CGRP and opioid peptides in NRM of rats. CGRP-like immunoreactivity was found at a concentration of 6.0+/-0. 77 pmol/g in NRM tissue of ten samples of rats, suggesting that it may contribute to physiological responses orchestrated by the NRM. The hindpaw withdrawal latency (HWL) to thermal and mechanical stimulation increased significantly after intra-NRM administration of 0.5 or 1 nmol of CGRP in rats, but not 0.25 nmol. The anti-nociceptive effect induced by CGRP was antagonized by following intra-NRM injection of 1 nmol of the CGRP receptor antagonist CGRP8-37. Furthermore, the CGRP-induced anti-nociceptive effect was attenuated by following intra-NRM administration of 6 nmol of naloxone. The results indicate that CGRP and its receptors play an important role in anti-nociception, and there is a possible interaction between CGRP and opioid peptides in NRM of rats.

  6. Electrolytic lesion of the nucleus raphe magnus reduced the antinociceptive effects of bilateral morphine microinjected into the nucleus cuneiformis in rats.

    Science.gov (United States)

    Haghparast, Abbas; Ordikhani-Seyedlar, Mehdi; Ziaei, Maryam

    2008-06-27

    Several lines of investigation show that the rostral ventromedial medulla is a critical relay for midbrain regions, including the nucleus cuneiformis (CnF), which control nociception at the spinal cord. There is some evidence that local stimulation or morphine administration into the CnF produces the effective analgesia through the nucleus raphe magnus (NRM). The present study tries to determine the effect of morphine-induced analgesia following microinjection into the CnF in the absence of NRM. Seven days after the cannulae implantation, morphine was microinjected bilaterally into the CnF at the doses of 0.25, 1, 2.5, 5, 7.5 and 10 microg/0.3 microl saline per side. The morphine-induced antinociceptive effect measured by tail-flick test at 30, 60, 90 and 120 min after microinjection. The results showed that bilateral microinjection of morphine into the CnF dose-dependently causes increase in tail-flick latency (TFL). The 50% effective dose of morphine was determined and microinjected into the CnF (2.5 microg/0.3 microl saline per side) in rats after NRM electrolytic lesion (1 mA, 30 s). Lesion of the NRM significantly decreased TFLs, 30 (Peffects through the opioid receptors in the CnF. It is also appeared that morphine-induced antinociception decreases following the NRM lesion but it seems that there are some other descending pain modulatory pathways that activate in the absence of NRM.

  7. Reversible inactivation and excitation of nucleus raphe magnus can modulate tail blood flow of male Wistar rats in response to hypothermia.

    Science.gov (United States)

    Malakouti, Seyed Mansour; Kourosh Arami, Masoomeh; Sarihi, Abdorahman; Hajizadeh, Sohrab; Behzadi, Gila; Shahidi, Siamak; Komaki, Alireza; Heshmatian, Behnam; Vahabian, Mehrangiz

    2008-10-01

    The nucleus raphe magnus (NRM) is involved in thermoregulatory processing. There is a correlation between changes in the firing rates of the cells in the NRM and the application of the peripheral thermal stimulus. we examined the effect of reversible inactivation and excitation of NRM on mechanisms involved in tail blood flow (TBF) regulation in hypothermia. Hypothermia was induced in Male Wistar rats and cannula was implanted above the NRM. To evaluate the effect of nucleus inactivation on TBF, the amount of TBF was measured by Laser Doppler in hypothermic rats, before and after lidocaine microinjection into NRM. TBF was also measured after glutamate microinjection to assess the effect of nucleus excitation in hypothermic rats. Results indicated that after dropping TBF by hypothermia, microinjection of lidocaine into NRM significantly decreased TBF from 54.43 +- 5.7 to 46.81 +- 3.4, whereas glutamate microinjection caused a significant increase from 44.194 +- 0.6 to 98 +- 10.0 CONCLUSION: These data suggest that NRM have thermoregulatory effect in response to hypothermia.

  8. Brief pup exposure induces Fos expression in the lateral habenula and serotonergic caudal dorsal raphe nucleus of paternally experienced male California mice (Peromyscus californicus).

    Science.gov (United States)

    de Jong, T R; Measor, K R; Chauke, M; Harris, B N; Saltzman, W

    2010-09-01

    Fathers play a substantial role in infant care in a small but significant number of mammalian species, including humans. However, the neural circuitry controlling paternal behavior is much less understood than its female counterpart. In order to characterize brain areas activated by paternal care, male California mice were separated from their female mate and litter for 3 h and then exposed to a pup or a control object (a glass pebble with the approximate size and oblong shape of a newborn pup) for 10 min. All males receiving a pup showed a strong paternal response towards it, whereas males receiving a pebble interacted with it only occasionally. Despite the clear behavioral differences, exposure to a pup did not increase Fos-like immunoreactivity (Fos-LIR) compared to a pebble in brain areas previously found to be associated with parental care, including the medial preoptic nucleus and medial bed nucleus of the stria terminalis. Pup exposure did, however, significantly increase Fos-LIR in the lateral habenula (LHb) and in predominantly serotonergic neurons in the caudal dorsal raphe nucleus (DRC), as compared to pebble exposure. Both the LHb and DRC are known to be involved in the behavioral responses to strong emotional stimuli; therefore, these areas might play a role in controlling parental behavior in male California mice. Copyright (c) 2010 IBRO. Published by Elsevier Ltd. All rights reserved.

  9. Creating corporate advantage.

    Science.gov (United States)

    Collis, D J; Montgomery, C A

    1998-01-01

    What differentiates truly great corporate strategies from the merely adequate? How can executives at the corporate level create tangible advantage for their businesses that makes the whole more than the sum of the parts? This article presents a comprehensive framework for value creation in the multibusiness company. It addresses the most fundamental questions of corporate strategy: What businesses should a company be in? How should it coordinate activities across businesses? What role should the corporate office play? How should the corporation measure and control performance? Through detailed case studies of Tyco International, Sharp, the Newell Company, and Saatchi and Saatchi, the authors demonstrate that the answers to all those questions are driven largely by the nature of a company's special resources--its assets, skills, and capabilities. These range along a continuum from the highly specialized at one end to the very general at the other. A corporation's location on the continuum constrains the set of businesses it should compete in and limits its choices about the design of its organization. Applying the framework, the authors point out the common mistakes that result from misaligned corporate strategies. Companies mistakenly enter businesses based on similarities in products rather than the resources that contribute to competitive advantage in each business. Instead of tailoring organizational structures and systems to the needs of a particular strategy, they create plain-vanilla corporate offices and infrastructures. The company examples demonstrate that one size does not fit all. One can find great corporate strategies all along the continuum.

  10. Creating sustainable performance.

    Science.gov (United States)

    Spreitzer, Gretchen; Porath, Christine

    2012-01-01

    What makes for sustainable individual and organizational performance? Employees who are thriving-not just satisfied and productive but also engaged in creating the future. The authors found that people who fit this description demonstrated 16% better overall performance, 125% less burnout, 32% more commitment to the organization, and 46% more job satisfaction than their peers. Thriving has two components: vitality, or the sense of being alive and excited, and learning, or the growth that comes from gaining knowledge and skills. Some people naturally build vitality and learning into their jobs, but most employees are influenced by their environment. Four mechanisms, none of which requires heroic effort or major resources, create the conditions for thriving: providing decision-making discretion, sharing information about the organization and its strategy, minimizing incivility, and offering performance feedback. Organizations such as Alaska Airlines, Zingerman's, Quicken Loans, and Caiman Consulting have found that helping people grow and remain energized at work is valiant on its own merits-but it can also boost performance in a sustainable way.

  11. Identification of a Novel EF-Loop in the N-terminus of TRPM2 Channel Involved in Calcium Sensitivity

    Directory of Open Access Journals (Sweden)

    Yuhuan Luo

    2018-06-01

    Full Text Available As an oxidative stress sensor, transient receptor potential melastatin 2 (TRPM2 channel is involved in many physiological and pathological processes including warmth sensing, ischemia injury, inflammatory diseases and diabetes. Intracellular calcium is critical for TRPM2 channel activation and the IQ-like motif in the N-terminus has been shown to be important by mediating calmodulin binding. Sequence analysis predicted two potential EF-loops in the N-terminus of TRPM2. Site-directed mutagenesis combining with functional assay showed that substitution with alanine of several residues, most of which are conserved in the typical EF-loop, including D267, D278, D288, and E298 dramatically reduced TRPM2 channel currents. By further changing the charges or side chain length of these conserved residues, our results indicate that the negative charge of D267 and the side chain length of D278 are critical for calcium-induced TRPM2 channel activation. G272I mutation also dramatically reduced the channel currents, suggesting that this site is critical for calcium-induced TRPM2 channel activation. Furthermore, D267A mutant dramatically reduced the currents induced by calcium alone compared with that by ADPR, indicating that D267 residue in D267–D278 motif is the most important site for calcium sensitivity of TRPM2. In addition, inside-out recordings showed that mutations at D267, G272, D278, and E298 had no effect on single-channel conductance. Taken together, our data indicate that D267–D278 motif in the N-terminus as a novel EF-loop is critical for calcium-induced TRPM2 channel activation.

  12. Creating With Carbon

    Science.gov (United States)

    2003-01-01

    A subsidiary of SI Diamond Technology, Inc., Applied Nanotech, of Austin, Texas, is creating a buzz among various technology firms and venture capital groups interested in the company s progressive research on carbon-related field emission devices, including carbon nanotubes, filaments of pure carbon less than one ten-thousandth the width of human hair. Since their discovery in 1991, carbon nanotubes have gained considerable attention due to their unique physical properties. For example, a single perfect carbon nanotube can range from 10 to 100 times stronger than steel, per unit weight. Recent studies also indicate that the nanotubes may be the best heat-conducting material in existence. These properties, combined with the ease of growing thin films or nanotubes by a variety of deposition techniques, make the carbon-based material one of the most desirable for cold field emission cathodes.

  13. A Reduced Risk of Infection with Plasmodium vivax and Clinical Protection against Malaria Are Associated with Antibodies against the N Terminus but Not the C Terminus of Merozoite Surface Protein 1†

    Science.gov (United States)

    Nogueira, Paulo Afonso; Piovesan Alves, Fabiana; Fernandez-Becerra, Carmen; Pein, Oliver; Rodrigues Santos, Neida; Pereira da Silva, Luiz Hildebrando; Plessman Camargo, Erney; del Portillo, Hernando A.

    2006-01-01

    Progress towards the development of a malaria vaccine against Plasmodium vivax, the most widely distributed human malaria parasite, will require a better understanding of the immune responses that confer clinical protection to patients in regions where malaria is endemic. The occurrence of clinical protection in P. vivax malaria in Brazil was first reported among residents of the riverine community of Portuchuelo, in Rondônia, western Amazon. We thus analyzed immune sera from this same human population to determine if naturally acquired humoral immune responses against the merozoite surface protein 1 of P. vivax, PvMSP1, could be associated with reduced risk of infection and/or clinical protection. Our results demonstrated that this association could be established with anti-PvMSP1 antibodies predominantly of the immunoglobulin G3 subclass directed against the N terminus but not against the C terminus, in spite of the latter being more immunogenic and capable of natural boosting. This is the first report of a prospective study of P. vivax malaria demonstrating an association of reduced risk of infection and clinical protection with antibodies against an antigen of this parasite. PMID:16622209

  14. Creating the living brand.

    Science.gov (United States)

    Bendapudi, Neeli; Bendapudi, Venkat

    2005-05-01

    It's easy to conclude from the literature and the lore that top-notch customer service is the province of a few luxury companies and that any retailer outside that rarefied atmosphere is condemned to offer mediocre service at best. But even companies that position themselves for the mass market can provide outstanding customer-employee interactions and profit from them, if they train employees to reflect the brand's core values. The authors studied the convenience store industry in depth and focused on two that have developed a devoted following: QuikTrip (QT) and Wawa. Turnover rates at QT and Wawa are 14% and 22% respectively, much lower than the typical rate in retail. The authors found six principles that both firms embrace to create a strong culture of customer service. Know what you're looking for: A focus on candidates' intrinsic traits allows the companies to hire people who will naturally bring the right qualities to the job. Make the most of talent: In mass-market retail, talent is generally viewed as a commodity, but that outlook becomes a self-fulfilling prophesy. Create pride in the brand: Service quality depends directly on employees' attachment to the brand. Build community: Wawa and QT have made concerted efforts to build customer loyalty through a sense of community. Share the business context: Employees need a clear understanding of how their company operates and how it defines success. Satisfy the soul: To win an employee's passionate engagement, a company must meet his or her needs for security, esteem, and justice.

  15. NMR spectroscopic and bioinformatic analyses of the LTBP1 C-terminus reveal a highly dynamic domain organisation.

    Directory of Open Access Journals (Sweden)

    Ian B Robertson

    Full Text Available Proteins from the LTBP/fibrillin family perform key structural and functional roles in connective tissues. LTBP1 forms the large latent complex with TGFβ and its propeptide LAP, and sequesters the latent growth factor to the extracellular matrix. Bioinformatics studies suggest the main structural features of the LTBP1 C-terminus are conserved through evolution. NMR studies were carried out on three overlapping C-terminal fragments of LTBP1, comprising four domains with characterised homologues, cbEGF14, TB3, EGF3 and cbEGF15, and three regions with no homology to known structures. The NMR data reveal that the four domains adopt canonical folds, but largely lack the interdomain interactions observed with homologous fibrillin domains; the exception is the EGF3-cbEGF15 domain pair which has a well-defined interdomain interface. (15N relaxation studies further demonstrate that the three interdomain regions act as flexible linkers, allowing a wide range of motion between the well-structured domains. This work is consistent with the LTBP1 C-terminus adopting a flexible "knotted rope" structure, which may facilitate cell matrix interactions, and the accessibility to proteases or other factors that could contribute to TGFβ activation.

  16. Physical interaction of junctophilin and the CaV1.1 C terminus is crucial for skeletal muscle contraction.

    Science.gov (United States)

    Nakada, Tsutomu; Kashihara, Toshihide; Komatsu, Masatoshi; Kojima, Katsuhiko; Takeshita, Toshikazu; Yamada, Mitsuhiko

    2018-04-24

    Close physical association of Ca V 1.1 L-type calcium channels (LTCCs) at the sarcolemmal junctional membrane (JM) with ryanodine receptors (RyRs) of the sarcoplasmic reticulum (SR) is crucial for excitation-contraction coupling (ECC) in skeletal muscle. However, the molecular mechanism underlying the JM targeting of LTCCs is unexplored. Junctophilin 1 (JP1) and JP2 stabilize the JM by bridging the sarcolemmal and SR membranes. Here, we examined the roles of JPs in localization and function of LTCCs. Knockdown of JP1 or JP2 in cultured myotubes inhibited LTCC clustering at the JM and suppressed evoked Ca 2+ transients without disrupting JM structure. Coimmunoprecipitation and GST pull-down assays demonstrated that JPs physically interacted with 12-aa residues in the proximal C terminus of the Ca V 1.1. A JP1 mutant lacking the C terminus including the transmembrane domain (JP1ΔCT) interacted with the sarcolemmal/T-tubule membrane but not the SR membrane. Expression of this mutant in adult mouse muscles in vivo exerted a dominant-negative effect on endogenous JPs, impairing LTCC-RyR coupling at triads without disrupting JM morphology, and substantially reducing Ca 2+ transients without affecting SR Ca 2+ content. Moreover, the contractile force of the JP1ΔCT-expressed muscle was dramatically reduced compared with the control. Taken together, JPs recruit LTCCs to the JM through physical interaction and ensure robust ECC at triads in skeletal muscle.

  17. Identification of a tetrameric assembly domain in the C terminus of heat-activated TRPV1 channels.

    Science.gov (United States)

    Zhang, Feng; Liu, Shuang; Yang, Fan; Zheng, Jie; Wang, KeWei

    2011-04-29

    Transient receptor potential (TRP) channels as cellular sensors are thought to function as tetramers. Yet, the molecular determinants governing channel multimerization remain largely elusive. Here we report the identification of a segment comprising 21 amino acids (residues 752-772 of mouse TRPV1) after the known TRP-like domain in the channel C terminus that functions as a tetrameric assembly domain (TAD). Purified recombinant C-terminal proteins of TRPV1-4, but not the N terminus, mediated the protein-protein interaction in an in vitro pulldown assay. Western blot analysis combined with electrophysiology and calcium imaging demonstrated that TAD exerted a robust dominant-negative effect on wild-type TRPV1. When fused with the membrane-tethered peptide Gap43, the TAD blocked the formation of stable homomultimers. Calcium imaging and current recordings showed that deletion of the TAD in a poreless TRPV1 mutant subunit suppressed its dominant-negative phenotype, confirming the involvement of the TAD in assembly of functional channels. Our findings suggest that the C-terminal TAD in TRPV1 channels functions as a domain that is conserved among TRPV1-4 and mediates a direct subunit-subunit interaction for tetrameric assembly.

  18. Creating an outcomes framework.

    Science.gov (United States)

    Doerge, J B

    2000-01-01

    Four constructs used to build a framework for outcomes management for a large midwestern tertiary hospital are described in this article. A system framework outlining a model of clinical integration and population management based in Steven Shortell's work is discussed. This framework includes key definitions of high-risk patients, target groups, populations and community. Roles for each level of population management and how they were implemented in the health care system are described. A point of service framework centered on seven dimensions of care is the next construct applied on each nursing unit. The third construct outlines the framework for role development. Three roles for nursing were created to implement strategies for target groups that are strategic disease categories; two of those roles are described in depth. The philosophy of nursing practice is centered on caring and existential advocacy. The final construct is the modification of the Dartmouth model as a common framework for outcomes. System applications of the scorecard and lessons learned in the 2-year process of implementation are shared

  19. Creating a disability mythology.

    Science.gov (United States)

    Brown, S E

    1992-01-01

    People with disabilities have, for the most part, failed to identify with each other as a group. This has been detrimental because it has built a sense of isolation when a camaraderie based upon existing commonalities could have been developed. During the past ten to twenty years, there has been a great deal of discussion about appropriate language to use when discussing disability issues. This discussion has been a part of a larger debate concerning the existence of a disability culture. I believe that there is indeed a disability culture and I am a proponent of identifying and passing on stories which contribute to that culture. I have chosen to use mythology to convey this message and have begun with a focus on heroes - people who do something out of the ordinary. It is contended that almost all people with disabilities have performed heroic activities because of the pervasive discrimination encountered by each individual with a disability. Creating a disability mythology is an attempt to recognize and promote heroes within the disabled community and to advocate the importance of telling other people how positive change has occurred through instances of individual heroism.

  20. Corticotropin-releasing Factor in the Rat Dorsal Raphe Nucleus Promotes Different Forms of Behavioral Flexibility Depending on Social Stress History.

    Science.gov (United States)

    Snyder, Kevin P; Hill-Smith, Tiffany E; Lucki, Irwin; Valentino, Rita J

    2015-10-01

    The stress-related neuropeptide, corticotropin-releasing factor (CRF) regulates the dorsal raphe nucleus-serotonin (DRN-5-HT) system during stress and this may underlie affective and cognitive dysfunctions that characterize stress-related psychiatric disorders. CRF acts on both CRF1 and CRF2 receptor subtypes in the DRN that exert opposing inhibitory and excitatory effects on DRN-5-HT neuronal activity and 5-HT forebrain release, respectively. The current study first assessed the cognitive effects of intra-DRN microinfusion of CRF or the selective CRF2 agonist, urocortin II in stress-naive rats on performance of an operant strategy set-shifting task that is mediated by the medial prefrontal cortex (mPFC). CRF (30 ng) facilitated strategy set-shifting performance, whereas higher doses of CRF and urocortin II that would interact with CRF2 were without effect, consistent with a CRF1-mediated action. This dose decreased 5-HT extracellular levels in the mPFC, further supporting a role for CRF1. The effects of CRF were then assessed in rats exposed to repeated social stress using the resident-intruder model. Repeated social stress shifted the CRF effect from facilitation of strategy set shifting to facilitation of reversal learning and this was most prominent in a subpopulation of rats that resist defeat. Notably, in this subpopulation of rats 5-HT neuronal responses to CRF have been demonstrated to shift from CRF1-mediated inhibition to CRF2-mediated excitation. Because 5-HT facilitates reversal learning, the present results suggest that stress-induced changes in the cellular effects of CRF in the DRN translate to changes in cognitive effects of CRF. Together, the results underscore the potential for stress history to shift cognitive processing through changes in CRF neurotransmission in the DRN and the association of this effect with coping strategy.

  1. Effects of cocaine history on postsynaptic GABA receptors on dorsal raphe serotonin neurons in a stress-induced relapse model in rats.

    Science.gov (United States)

    Li, Chen; Kirby, Lynn G

    2016-01-01

    The serotonin (5-hydroxytryptamine, 5-HT) system plays an important role in stress-related psychiatric disorders and substance abuse. Stressors and stress hormones can inhibit the dorsal raphe nucleus (DRN)-5-HT system, which composes the majority of forebrain-projecting 5-HT. This inhibition is mediated via stimulation of GABA synaptic activity at DRN-5-HT neurons. Using swim stress-induced reinstatement of morphine conditioned place-preference, recent data from our laboratory indicate that morphine history sensitizes DRN-5-HT neurons to GABAergic inhibitory effects of stress. Moreover, GABAA receptor-mediated inhibition of the serotonergic DRN is required for this reinstatement. In our current experiment, we tested the hypothesis that GABAergic sensitization of DRN-5-HT neurons is a neuroadaptation elicited by multiple classes of abused drugs across multiple models of stress-induced relapse by applying a chemical stressor (yohimbine) to induce reinstatement of previously extinguished cocaine self-administration in Sprague-Dawley rats. Whole-cell patch-clamp recordings of GABA synaptic activity in DRN-5-HT neurons were conducted after the reinstatement. Behavioral data indicate that yohimbine triggered reinstatement of cocaine self-administration. Electrophysiology data indicate that 5-HT neurons in the cocaine group exposed to yohimbine had increased amplitude of inhibitory postsynaptic currents compared to yoked-saline controls exposed to yohimbine or unstressed animals in both drug groups. These data, together with previous findings, indicate that interaction between psychostimulant or opioid history and chemical or physical stressors may increase postsynaptic GABA receptor density and/or sensitivity in DRN-5-HT neurons. Such mechanisms may result in serotonergic hypofunction and consequent dysphoric mood states which confer vulnerability to stress-induced drug reinstatement. Copyright © 2015 Elsevier B.V. and ECNP. All rights reserved.

  2. Excitatory amino acid receptor blockade within the caudal pressor area and rostral ventrolateral medulla alters cardiovascular responses to nucleus raphe obscurus stimulation in rats

    Directory of Open Access Journals (Sweden)

    Silva N.F.

    2002-01-01

    Full Text Available Pressor responses elicited by stimulation of the nucleus raphe obscurus (NRO depend on the integrity of the rostral ventrolateral medulla (RVLM. Therefore, to test the participation of excitatory amino acid (EAA receptors in the cardiovascular responses evoked by NRO stimulation (1 ms, 100 Hz, 40-70 µA, for 10 s, the EAA antagonist kynurenic acid (Kyn was microinjected at different sites in the ventrolateral medullar surface (2.7 nmol/200 nl of male Wistar rats (270-320 g, N = 39 and NRO stimulation was repeated. The effects of NRO stimulation were: hypertension (deltaMAP = +43 ± 1 mmHg, P<0.01, bradycardia (deltaHR = -30 ± 7 bpm, P<0.01 and apnea. Bilateral microinjection of Kyn into the RVLM, which did not change baseline parameters, almost abolished the bradycardia induced by NRO stimulation (deltaHR = -61 ± 3 before vs -2 ± 3 bpm after Kyn, P<0.01, N = 7. Unilateral microinjection of Kyn into the CVLM did not change baseline parameters or reduce the pressor response to NRO stimulation (deltaMAP = +46 ± 5 before vs +48 ± 5 mmHg after Kyn, N = 6. Kyn bilaterally microinjected into the caudal pressor area reduced blood pressure and heart rate and almost abolished the pressor response to NRO stimulation (deltaMAP = +46 ± 4 mmHg before vs +4 ± 2 mmHg after Kyn, P<0.01, N = 7. These results indicate that EAA receptors on the medullary ventrolateral surface play a role in the modulation of the cardiovascular responses induced by NRO stimulation, and also suggest that the RVLM participates in the modulation of heart rate responses and that the caudal pressor area modulates the pressor response following NRO stimulation.

  3. Role of glutamatergic receptors located in the nucleus raphe magnus on antinociceptive effect of morphine microinjected into the nucleus cuneiformis of rat.

    Science.gov (United States)

    Haghparast, Abbas; Soltani-Hekmat, Ava; Khani, Abbas; Komaki, Alireza

    2007-10-29

    Neurons in the nucleus cuneiformis (CnF), located just ventrolateral to the periaqueductal gray, project to medullary nucleus raphe magnus (NRM), which is a key medullary relay for descending pain modulation and is critically involved in opioid-induced analgesia. Previous studies have shown that antinociceptive response of CnF-microinjected morphine can be modulated by the specific subtypes of glutamatergic receptors within the CnF. In this study, we evaluated the role of NMDA and kainate/AMPA receptors that are widely distributed within the NRM on morphine-induced antinociception elicited from the CnF. Hundred and five male Wistar rats weighing 250-300 g were used. Morphine (10, 20 and 40 microg) and NMDA receptor antagonist, MK-801 (10 microg) or kainate/AMPA receptor antagonist, DNQX (0.5 microg) in 0.5 microl saline were stereotaxically microinjected into the CnF and NRM, respectively. The latency of tail-flick response was measured at set intervals (2, 7, 12, 17, 22, 27 min after microinjection) by using an automated tail-flick analgesiometer. The results showed that morphine microinjection into the CnF dose-dependently causes increase in tail-flick latency (TFL). MK-801 microinjected into the NRM, just 1 min before morphine injection into the CnF, significantly attenuated antinociceptive effects of morphine. On the other hand, DNQX microinjected into the NRM, significantly increased TFL after local application of morphine into the CnF. We suggest that morphine related antinociceptive effect elicited from the CnF is mediated, in part, by NMDA receptor at the level of the NRM whereas kainite/AMPA receptor has a net inhibitory influence at the same pathway.

  4. Creating a Social World

    Science.gov (United States)

    Kendler, Kenneth S.; Jacobson, Kristen C.; Gardner, Charles O.; Gillespie, Nathan; Aggen, Steven A.; Prescott, Carol A.

    2014-01-01

    Context Peer-group deviance is strongly associated with externalizing behaviors. We have limited knowledge of the sources of individual differences in peer-group deviance. Objective To clarify genetic and environmental contributions to peer-group deviance in twins from mid-childhood through early adulthood. Design Retrospective assessments using a life-history calendar. Analysis by biometric growth curves. Setting General community. Participants Members of male-male pairs from the population-based Virginia Twin Registry personally interviewed in 1998–2004 (n=1802). Main Outcome Measure Self-reported peer-group deviance at ages 8 to 11, 12 to 14, 15 to 17, 18 to 21, and 22 to 25 years. Results Mean and variance of peer-group deviance increased substantially with age. Genetic effects on peer-group deviance showed a strong and steady increase over time. Family environment generally declined in importance over time. Individual-specific environmental influences on peer-group deviance levels were stable in the first 3 age periods and then increased as most twins left home. When standardized, the heritability of peer-group deviance is approximately 30% at ages 8 to 11 years and rises to approximately 50% across the last 3 time periods. Both genes and shared environment contributed to individual differences in the developmental trajectory of peer-group deviance. However, while the correlation between childhood peer-group deviance levels and the subsequent slope of peer-group deviance over time resulting from genetic factors was positive, the same relationship resulting from shared environmental factors was negative. Conclusions As male twins mature and create their own social worlds, genetic factors play an increasingly important role in their choice of peers, while shared environment becomes less influential. The individual specific environment increases in importance when individuals leave home. Individuals who have deviant peers in childhood, as a result of genetic vs

  5. Creating a family health history

    Science.gov (United States)

    ... this page: //medlineplus.gov/ency/patientinstructions/000947.htm Creating a family health history To use the sharing ... Many health problems tend to run in families. Creating a family history can help you and your ...

  6. Brca2 C-terminus interacts with Rad51 and contributes to nuclear forcus formation in double-strand break repair of DNA

    International Nuclear Information System (INIS)

    Ochiai, Kazuhiko; Morimatsu, Masami; Yoshikawa, Yasunaga; Syuto, Bunei; Hashizume, Kazuyoshi

    2004-01-01

    In humans and mice, the interaction between the breast cancer susceptibility protein, Brca2, and Rad51 recombinase is essential for DNA repair by homologous recombination, the failure of this process can predispose to cancer. Cells with mutated Brca2 are hypersensitive to ionizing radiation (IR) and exhibit defective DNA repair. Using yeast and mammalian two-hybrid assays, we demonstrate that canine Rad51 protein interacts specifically with the C-terminus of canine Brca2. In support of the biological significance of this interaction, we found that radiation-induced focus formation of Rad51 in COS-7 cells was compromised by forced expression of the C-terminus of canine Brca2. A similar result was obtained for the murine C-terminus. These data suggest that the C-terminal domain of canine Brca2 functions to bind Rad51 and that this domain contributes to the IR-induced assembly of the Rad51 complex in vivo. (author)

  7. Simulating ice thickness and velocity evolution of Upernavik Isstrøm 1849-2012 by forcing prescribed terminus positions in ISSM

    Science.gov (United States)

    Haubner, Konstanze; Box, Jason E.; Schlegel, Nicole J.; Larour, Eric Y.; Morlighem, Mathieu; Solgaard, Anne M.; Kjeldsen, Kristian K.; Larsen, Signe H.; Rignot, Eric; Dupont, Todd K.; Kjær, Kurt H.

    2018-04-01

    Tidewater glacier velocity and mass balance are known to be highly responsive to terminus position change. Yet it remains challenging for ice flow models to reproduce observed ice margin changes. Here, using the Ice Sheet System Model (ISSM; Larour et al. 2012), we simulate the ice velocity and thickness changes of Upernavik Isstrøm (north-western Greenland) by prescribing a collection of 27 observed terminus positions spanning 164 years (1849-2012). The simulation shows increased ice velocity during the 1930s, the late 1970s and between 1995 and 2012 when terminus retreat was observed along with negative surface mass balance anomalies. Three distinct mass balance states are evident in the reconstruction: (1849-1932) with near zero mass balance, (1932-1992) with ice mass loss dominated by ice dynamical flow, and (1998-2012), when increased retreat and negative surface mass balance anomalies led to mass loss that was twice that of any earlier period. Over the multi-decadal simulation, mass loss was dominated by thinning and acceleration responsible for 70 % of the total mass loss induced by prescribed change in terminus position. The remaining 30 % of the total ice mass loss resulted directly from prescribed terminus retreat and decreasing surface mass balance. Although the method can not explain the cause of glacier retreat, it enables the reconstruction of ice flow and geometry during 1849-2012. Given annual or seasonal observed terminus front positions, this method could be a useful tool for evaluating simulations investigating the effect of calving laws.

  8. The C-terminus hot spot region helps in the fibril formation of bacteriophage-associated hyaluronate lyase (HylP2).

    Science.gov (United States)

    Shukla, Harish; Singh, Sudhir Kumar; Singh, Amit Kumar; Mitra, Kalyan; Akhtar, Md Sohail

    2015-09-23

    The bacteriophage encoded hyaluronate lyases (HylP and HylP2) degrade hyaluronan and other glycosaminoglycans. HylP2 forms a functional fibril under acidic conditions in which its N-terminus is proposed to form the fibrillar core, leading to nucleation and acceleration of fibril formation. Here we report the presence of a hot spot region (A144GVVVY149) towards the carboxy terminus of HylP2, essential for the acceleration of fibril formation. The 'hot spot' is observed to be inherently mutated for valines (A178AMVMY183) in case of HylP. The N- terminal swapped chimeras between these phage HLs ((N)HylP2(C)HylP and (N)HylP(C)HylP2) or HylP did not form fibrils at acidic pH. However, seeding of prefibrils of HylP2 recompensed nucleation and led to fibrillation in (N)HylP(C)HylP2. The V147A mutation in the 'hot spot' region abolished fibril formation in HylP2. The M179V and M181V double mutations in the 'hot spot' region of HylP led to fibrillation with the seeding of prefibrils. It appears that fibrillation in HylP2 even though is initiated by the N-terminus, is accelerated by the conserved 'hot spot' region in the C-terminus. A collagenous (Gly-X-Y)10 motif in the N-terminus and a mutated 'hot spot' region in the C-terminus of HylP affect fibrillar nucleation and acceleration respectively.

  9. Phosphorylation and cellular function of the human Rpa2 N-terminus in the budding yeast Saccharomyces cerevisiae.

    Science.gov (United States)

    Ghospurkar, Padmaja L; Wilson, Timothy M; Liu, Shengqin; Herauf, Anna; Steffes, Jenna; Mueller, Erica N; Oakley, Gregory G; Haring, Stuart J

    2015-02-01

    Maintenance of genome integrity is critical for proper cell growth. This occurs through accurate DNA replication and repair of DNA lesions. A key factor involved in both DNA replication and the DNA damage response is the heterotrimeric single-stranded DNA (ssDNA) binding complex Replication Protein A (RPA). Although the RPA complex appears to be structurally conserved throughout eukaryotes, the primary amino acid sequence of each subunit can vary considerably. Examination of sequence differences along with the functional interchangeability of orthologous RPA subunits or regions could provide insight into important regions and their functions. This might also allow for study in simpler systems. We determined that substitution of yeast Replication Factor A (RFA) with human RPA does not support yeast cell viability. Exchange of a single yeast RFA subunit with the corresponding human RPA subunit does not function due to lack of inter-species subunit interactions. Substitution of yeast Rfa2 with domains/regions of human Rpa2 important for Rpa2 function (i.e., the N-terminus and the loop 3-4 region) supports viability in yeast cells, and hybrid proteins containing human Rpa2 N-terminal phospho-mutations result in similar DNA damage phenotypes to analogous yeast Rfa2 N-terminal phospho-mutants. Finally, the human Rpa2 N-terminus (NT) fused to yeast Rfa2 is phosphorylated in a manner similar to human Rpa2 in human cells, indicating that conserved kinases recognize the human domain in yeast. The implication is that budding yeast represents a potential model system for studying not only human Rpa2 N-terminal phosphorylation, but also phosphorylation of Rpa2 N-termini from other eukaryotic organisms. Copyright © 2014 The Authors. Published by Elsevier Inc. All rights reserved.

  10. Regulating repression: roles for the sir4 N-terminus in linker DNA protection and stabilization of epigenetic states.

    Directory of Open Access Journals (Sweden)

    Stephanie Kueng

    Full Text Available Silent information regulator proteins Sir2, Sir3, and Sir4 form a heterotrimeric complex that represses transcription at subtelomeric regions and homothallic mating type (HM loci in budding yeast. We have performed a detailed biochemical and genetic analysis of the largest Sir protein, Sir4. The N-terminal half of Sir4 is dispensable for SIR-mediated repression of HM loci in vivo, except in strains that lack Yku70 or have weak silencer elements. For HM silencing in these cells, the C-terminal domain (Sir4C, residues 747-1,358 must be complemented with an N-terminal domain (Sir4N; residues 1-270, expressed either independently or as a fusion with Sir4C. Nonetheless, recombinant Sir4C can form a complex with Sir2 and Sir3 in vitro, is catalytically active, and has sedimentation properties similar to a full-length Sir4-containing SIR complex. Sir4C-containing SIR complexes bind nucleosomal arrays and protect linker DNA from nucleolytic digestion, but less effectively than wild-type SIR complexes. Consistently, full-length Sir4 is required for the complete repression of subtelomeric genes. Supporting the notion that the Sir4 N-terminus is a regulatory domain, we find it extensively phosphorylated on cyclin-dependent kinase consensus sites, some being hyperphosphorylated during mitosis. Mutation of two major phosphoacceptor sites (S63 and S84 derepresses natural subtelomeric genes when combined with a serendipitous mutation (P2A, which alone can enhance the stability of either the repressed or active state. The triple mutation confers resistance to rapamycin-induced stress and a loss of subtelomeric repression. We conclude that the Sir4 N-terminus plays two roles in SIR-mediated silencing: it contributes to epigenetic repression by stabilizing the SIR-mediated protection of linker DNA; and, as a target of phosphorylation, it can destabilize silencing in a regulated manner.

  11. Regulation of Hippocampal 5-HT Release by P2X7 Receptors in Response to Optogenetic Stimulation of Median Raphe Terminals of Mice

    Directory of Open Access Journals (Sweden)

    Flóra Gölöncsér

    2017-10-01

    Full Text Available Serotonergic and glutamatergic neurons of median raphe region (MRR play a pivotal role in the modulation of affective and cognitive functions. These neurons synapse both onto themselves and remote cortical areas. P2X7 receptors (P2rx7 are ligand gated ion channels expressed by central presynaptic excitatory nerve terminals and involved in the regulation of neurotransmitter release. P2rx7s are implicated in various neuropsychiatric conditions such as schizophrenia and depression. Here we investigated whether 5-HT release released from the hippocampal terminals of MRR is subject to modulation by P2rx7s. To achieve this goal, an optogenetic approach was used to selectively activate subpopulation of serotonergic terminals derived from the MRR locally, and one of its target area, the hippocampus. Optogenetic activation of neurons in the MRR with 20 Hz was correlated with freezing and enhanced locomotor activity of freely moving mice and elevated extracellular levels of 5-HT, glutamate but not GABA in vivo. Similar optical stimulation (OS significantly increased [3H]5-HT and [3H]glutamate release in acute MRR and hippocampal slices. We examined spatial and temporal patterns of [3H]5-HT release and the interaction between the serotonin and glutamate systems. Whilst [3H]5-HT release from MRR neurons was [Ca2+]o-dependent and sensitive to TTX, CNQX and DL-AP-5, release from hippocampal terminals was not affected by the latter drugs. Hippocampal [3H]5-HT released by electrical but not OS was subject to modulation by 5- HT1B/D receptors agonist sumatriptan (1 μM, whereas the selective 5-HT1A agonist buspirone (0.1 μM was without effect. [3H]5-HT released by electrical and optical stimulation was decreased in mice genetically deficient in P2rx7s, and after perfusion with selective P2rx7 antagonists, JNJ-47965567 (0.1 μM, and AZ-10606120 (0.1 μM. Optical and electrical stimulation elevated the extracellular level of ATP. Our results demonstrate for the

  12. A Stretch of 17 Amino Acids in the Prosaposin C Terminus Is Critical for Its Binding to Sortilin and Targeting to Lysosomes

    Science.gov (United States)

    Yuan, Libin; Morales, Carlos R.

    2010-01-01

    Prosaposin, the precursor of four lysosomal cofactors required for the hydrolysis of sphingolipids, is transported to the lysosomes via the alternative receptor, sortilin. In this study, we identified a specific domain of 17 amino acids within the C terminus of prosaposin involved in binding to this sorting receptor. We generated six prosaposin deletion constructs and examined the effect of truncation by coimmunoprecipitation and confocal microscopy. The experiments revealed that the first half of the prosaposin C terminus (aa 524–540), containing a saposin-like motif, was required and necessary to bind sortilin and to transport it to the lysosomes. Based on this result, we introduced twelve site-directed point mutations within the first half of the C terminus. Although the interaction of prosaposin with sortilin was pH dependent, the mutation of hydrophilic amino acids that usually modulate pH-dependent protein interactions did not affect the binding of prosaposin to sortilin. Conversely, a tryptophan (W530) and two cysteines (C528 and C536) were essential for its interaction with sortilin and for its transport to the lysosomes. In conclusion, our investigation demonstrates that a saposin-like motif within the first half of the prosaposin C terminus contains the sortilin recognition site. (J Histochem Cytochem 58:287–300, 2010) PMID:19934382

  13. Creating

    DEFF Research Database (Denmark)

    Andersen, Kristina Vaarst; Lorenzen, Mark; Laursen, Stine

    2012-01-01

    This unique book reveals the procedural aspects of knowledge-based urban planning, development and assessment. Concentrating on major knowledge city building processes, and providing state-of-the-art experiences and perspectives, this important compendium explores innovative models, approaches an...

  14. The C-terminus of the B-chain of human insulin-like peptide 5 is critical for cognate RXFP4 receptor activity.

    Science.gov (United States)

    Patil, Nitin A; Bathgate, Ross A D; Kocan, Martina; Ang, Sheng Yu; Tailhades, Julien; Separovic, Frances; Summers, Roger; Grosse, Johannes; Hughes, Richard A; Wade, John D; Hossain, Mohammed Akhter

    2016-04-01

    Insulin-like peptide 5 (INSL5) is an orexigenic peptide hormone belonging to the relaxin family of peptides. It is expressed primarily in the L-cells of the colon and has a postulated key role in regulating food intake. Its G protein-coupled receptor, RXFP4, is a potential drug target for treating obesity and anorexia. We studied the effect of modification of the C-terminus of the A and B-chains of human INSL5 on RXFP4 binding and activation. Three variants of human INSL5 were prepared using solid phase peptide synthesis and subsequent sequential regioselective disulfide bond formation. The peptides were synthesized as C-terminal acids (both A- and B-chains with free C-termini, i.e., the native form), amides (both chains as the C-terminal amide) and one analog with the C-terminus of its A-chain as the amide and the C-terminus of the B-chain as the acid. The results showed that C-terminus of the B-chain is more important than that of the A-chain for RXFP4 binding and activity. Amidation of the A-chain C-terminus does not have any effect on the INSL5 activity. The difference in RXFP4 binding and activation between the three peptides is believed to be due to electrostatic interaction of the free carboxylate of INSL5 with a positively charged residue (s), either situated within the INSL5 molecule itself or in the receptor extracellular loops.

  15. The N-Terminus of the Floral Arabidopsis TGA Transcription Factor PERIANTHIA Mediates Redox-Sensitive DNA-Binding.

    Directory of Open Access Journals (Sweden)

    Nora Gutsche

    Full Text Available The Arabidopsis TGA transcription factor (TF PERIANTHIA (PAN regulates the formation of the floral organ primordia as revealed by the pan mutant forming an abnormal pentamerous arrangement of the outer three floral whorls. The Arabidopsis TGA bZIP TF family comprises 10 members, of which PAN and TGA9/10 control flower developmental processes and TGA1/2/5/6 participate in stress-responses. For the TGA1 protein it was shown that several cysteines can be redox-dependently modified. TGA proteins interact in the nucleus with land plant-specific glutaredoxins, which may alter their activities posttranslationally. Here, we investigated the DNA-binding of PAN to the AAGAAT motif under different redox-conditions. The AAGAAT motif is localized in the second intron of the floral homeotic regulator AGAMOUS (AG, which controls stamen and carpel development as well as floral determinacy. Whereas PAN protein binds to this regulatory cis-element under reducing conditions, the interaction is strongly reduced under oxidizing conditions in EMSA studies. The redox-sensitive DNA-binding is mediated via a special PAN N-terminus, which is not present in other Arabidopsis TGA TFs and comprises five cysteines. Two N-terminal PAN cysteines, Cys68 and Cys87, were shown to form a disulfide bridge and Cys340, localized in a C-terminal putative transactivation domain, can be S-glutathionylated. Comparative land plant analyses revealed that the AAGAAT motif exists in asterid and rosid plant species. TGA TFs with N-terminal extensions of variable length were identified in all analyzed seed plants. However, a PAN-like N-terminus exists only in the rosids and exclusively Brassicaceae homologs comprise four to five of the PAN N-terminal cysteines. Redox-dependent modifications of TGA cysteines are known to regulate the activity of stress-related TGA TFs. Here, we show that the N-terminal PAN cysteines participate in a redox-dependent control of the PAN interaction with a highly

  16. Novel P2 promoter-derived HNF4α isoforms with different N-terminus generated by alternate exon insertion

    International Nuclear Information System (INIS)

    Huang, Jianmin; Levitsky, Lynne L.; Rhoads, David B.

    2009-01-01

    Hepatocyte nuclear factor 4α (HNF4α) is a critical transcription factor for pancreas and liver development and functions in islet β cells to maintain glucose homeostasis. Mutations in the human HNF4A gene lead to maturity onset diabetes of the young (MODY1) and polymorphisms are associated with increased risk for type 2 diabetes mellitus (T2DM). Expression of six HNF4α variants, three each from two developmentally regulated promoters, has been firmly established. We have now detected a new set of HNF4α variants designated HNF4α10-12 expressed from distal promoter P2. These variants, generated by inclusion of previously undetected exon 1E (human = 222 nt, rodent = 136 nt) following exon 1D have an altered N-terminus but identical remaining reading frame. HNF4α10-α12 are expressed in pancreatic islets (and liver) and exhibit transactivation potentials similar to the corresponding α7-α9 isoforms. DNA-binding analyses implied much higher protein levels of HNF4α10-α12 in liver than expected from the RT-PCR data. Our results provide evidence for a more complex expression pattern of HNF4α than previously appreciated. We recommend inclusion of exon 1E and nearby DNA sequences in screening for HNF4α mutations and polymorphisms in genetic analyses of MODY1 and T2DM.

  17. The human ACC2 CT-domain C-terminus is required for full functionality and has a novel twist

    Energy Technology Data Exchange (ETDEWEB)

    Madauss, Kevin P. [Department of Computational and Structural Chemistry, GlaxoSmithKline Inc., Five Moore Drive, Research Triangle Park, NC 27709 (United States); Burkhart, William A.; Consler, Thomas G. [Department of Biochemical Reagents and Assay Development, GlaxoSmithKline Inc., Five Moore Drive, Research Triangle Park, NC 27709 (United States); Cowan, David J. [Department of Chemistry in the Center for Excellence in Metabolic Pathways Drug Discovery, GlaxoSmithKline Inc., Five Moore Drive, Research Triangle Park, NC 27709 (United States); Gottschalk, William K. [Institute for Genome Sciences and Policy and Department of Medicine, Division of Neurology, Duke University, Durham, NC 27708 (United States); Miller, Aaron B. [Department of Computational and Structural Chemistry, GlaxoSmithKline Inc., Five Moore Drive, Research Triangle Park, NC 27709 (United States); Short, Steven A. [Department of Biochemical Reagents and Assay Development, GlaxoSmithKline Inc., Five Moore Drive, Research Triangle Park, NC 27709 (United States); Tran, Thuy B. [Department of Physiology, UNC School of Medicine, University of North Carolina, Chapel Hill, NC 27515 (United States); Williams, Shawn P., E-mail: shawn.p.williams@gsk.com [Department of Computational and Structural Chemistry, GlaxoSmithKline Inc., Five Moore Drive, Research Triangle Park, NC 27709 (United States)

    2009-05-01

    The use of biophysical assays permitted the identification of a specific human ACC2 carboxyl transferase (CT) domain mutant that binds inhibitors and crystallizes in their presence. This mutant led to determination of the human ACC2 CT domain–CP-640186 complex crystal structure, which revealed differences in the inhibitor conformation from the yeast protein complex that are caused by differing residues in the binding pocket. Inhibition of acetyl-CoA carboxylase (ACC) may prevent lipid-induced insulin resistance and type 2 diabetes, making the enzyme an attractive pharmaceutical target. Although the enzyme is highly conserved amongst animals, only the yeast enzyme structure is available for rational drug design. The use of biophysical assays has permitted the identification of a specific C-terminal truncation of the 826-residue human ACC2 carboxyl transferase (CT) domain that is both functionally competent to bind inhibitors and crystallizes in their presence. This C-terminal truncation led to the determination of the human ACC2 CT domain–CP-640186 complex crystal structure, which revealed distinctions from the yeast-enzyme complex. The human ACC2 CT-domain C-terminus is comprised of three intertwined α-helices that extend outwards from the enzyme on the opposite side to the ligand-binding site. Differences in the observed inhibitor conformation between the yeast and human structures are caused by differing residues in the binding pocket.

  18. Novel P2 promoter-derived HNF4{alpha} isoforms with different N-terminus generated by alternate exon insertion

    Energy Technology Data Exchange (ETDEWEB)

    Huang, Jianmin, E-mail: jmhuang@partners.org [Pediatric Endocrine Unit, MassGeneral Hospital for Children and Harvard Medical School, Boston, Massachusetts, 02114-2696 (United States); Levitsky, Lynne L. [Pediatric Endocrine Unit, MassGeneral Hospital for Children and Harvard Medical School, Boston, Massachusetts, 02114-2696 (United States); Rhoads, David B., E-mail: rhoads@helix.mgh.harvard.edu [Pediatric Endocrine Unit, MassGeneral Hospital for Children and Harvard Medical School, Boston, Massachusetts, 02114-2696 (United States)

    2009-04-15

    Hepatocyte nuclear factor 4{alpha} (HNF4{alpha}) is a critical transcription factor for pancreas and liver development and functions in islet {beta} cells to maintain glucose homeostasis. Mutations in the human HNF4A gene lead to maturity onset diabetes of the young (MODY1) and polymorphisms are associated with increased risk for type 2 diabetes mellitus (T2DM). Expression of six HNF4{alpha} variants, three each from two developmentally regulated promoters, has been firmly established. We have now detected a new set of HNF4{alpha} variants designated HNF4{alpha}10-12 expressed from distal promoter P2. These variants, generated by inclusion of previously undetected exon 1E (human = 222 nt, rodent = 136 nt) following exon 1D have an altered N-terminus but identical remaining reading frame. HNF4{alpha}10-{alpha}12 are expressed in pancreatic islets (and liver) and exhibit transactivation potentials similar to the corresponding {alpha}7-{alpha}9 isoforms. DNA-binding analyses implied much higher protein levels of HNF4{alpha}10-{alpha}12 in liver than expected from the RT-PCR data. Our results provide evidence for a more complex expression pattern of HNF4{alpha} than previously appreciated. We recommend inclusion of exon 1E and nearby DNA sequences in screening for HNF4{alpha} mutations and polymorphisms in genetic analyses of MODY1 and T2DM.

  19. GAIP interacting protein C-terminus regulates autophagy and exosome biogenesis of pancreatic cancer through metabolic pathways.

    Directory of Open Access Journals (Sweden)

    Santanu Bhattacharya

    Full Text Available GAIP interacting protein C terminus (GIPC is known to play an important role in a variety of physiological and disease states. In the present study, we have identified a novel role for GIPC as a master regulator of autophagy and the exocytotic pathways in cancer. We show that depletion of GIPC-induced autophagy in pancreatic cancer cells, as evident from the upregulation of the autophagy marker LC3II. We further report that GIPC regulates cellular trafficking pathways by modulating the secretion, biogenesis, and molecular composition of exosomes. We also identified the involvement of GIPC on metabolic stress pathways regulating autophagy and microvesicular shedding, and observed that GIPC status determines the loading of cellular cargo in the exosome. Furthermore, we have shown the overexpression of the drug resistance gene ABCG2 in exosomes from GIPC-depleted pancreatic cancer cells. We also demonstrated that depletion of GIPC from cancer cells sensitized them to gemcitabine treatment, an avenue that can be explored as a potential therapeutic strategy to overcome drug resistance in cancer.

  20. Purification method for recombinant proteins based on a fusion between the target protein and the C-terminus of calmodulin

    Science.gov (United States)

    Schauer-Vukasinovic, Vesna; Deo, Sapna K.; Daunert, Sylvia

    2002-01-01

    Calmodulin (CaM) was used as an affinity tail to facilitate the purification of the green fluorescent protein (GFP), which was used as a model target protein. The protein GFP was fused to the C-terminus of CaM, and a factor Xa cleavage site was introduced between the two proteins. A CaM-GFP fusion protein was expressed in E. coli and purified on a phenothiazine-derivatized silica column. CaM binds to the phenothiazine on the column in a Ca(2+)-dependent fashion and it was, therefore, used as an affinity tail for the purification of GFP. The fusion protein bound to the affinity column was then subjected to a proteolytic digestion with factor Xa. Pure GFP was eluted with a Ca(2+)-containing buffer, while CaM was eluted later with a buffer containing the Ca(2+)-chelating agent EGTA. The purity of the isolated GFP was verified by SDS-PAGE, and the fluorescence properties of the purified GFP were characterized.

  1. The Disordered C-Terminus of Yeast Hsf1 Contains a Cryptic Low-Complexity Amyloidogenic Region

    Directory of Open Access Journals (Sweden)

    Jordi Pujols

    2018-05-01

    Full Text Available Response mechanisms to external stress rely on networks of proteins able to activate specific signaling pathways to ensure the maintenance of cell proteostasis. Many of the proteins mediating this kind of response contain intrinsically disordered regions, which lack a defined structure, but still are able to interact with a wide range of clients that modulate the protein function. Some of these interactions are mediated by specific short sequences embedded in the longer disordered regions. Because the physicochemical properties that promote functional and abnormal interactions are similar, it has been shown that, in globular proteins, aggregation-prone and binding regions tend to overlap. It could be that the same principle applies for disordered protein regions. In this context, we show here that a predicted low-complexity interacting region in the disordered C-terminus of the stress response master regulator heat shock factor 1 (Hsf1 protein corresponds to a cryptic amyloid region able to self-assemble into fibrillary structures resembling those found in neurodegenerative disorders.

  2. The Disordered C-Terminus of Yeast Hsf1 Contains a Cryptic Low-Complexity Amyloidogenic Region.

    Science.gov (United States)

    Pujols, Jordi; Santos, Jaime; Pallarès, Irantzu; Ventura, Salvador

    2018-05-06

    Response mechanisms to external stress rely on networks of proteins able to activate specific signaling pathways to ensure the maintenance of cell proteostasis. Many of the proteins mediating this kind of response contain intrinsically disordered regions, which lack a defined structure, but still are able to interact with a wide range of clients that modulate the protein function. Some of these interactions are mediated by specific short sequences embedded in the longer disordered regions. Because the physicochemical properties that promote functional and abnormal interactions are similar, it has been shown that, in globular proteins, aggregation-prone and binding regions tend to overlap. It could be that the same principle applies for disordered protein regions. In this context, we show here that a predicted low-complexity interacting region in the disordered C-terminus of the stress response master regulator heat shock factor 1 (Hsf1) protein corresponds to a cryptic amyloid region able to self-assemble into fibrillary structures resembling those found in neurodegenerative disorders.

  3. Cloning and characterization of carboxyl terminus of heat shock cognate 70-interacting protein gene from the silkworm, Bombyx mori.

    Science.gov (United States)

    Ohsawa, Takeshi; Fujimoto, Shota; Tsunakawa, Akane; Shibano, Yuka; Kawasaki, Hideki; Iwanaga, Masashi

    2016-11-01

    Carboxyl terminus of heat shock cognate 70-interacting protein (CHIP) is an evolutionarily conserved E3 ubiquitin ligase across different eukaryotic species and is known to play a key role in protein quality control. CHIP has two distinct functional domains, an N-terminal tetratricopeptide repeat (TPR) and a C-terminal U-box domain, which are required for the ubiquitination of numerous labile client proteins that are chaperoned by heat shock proteins (HSPs) and heat shock cognate proteins (HSCs). During our screen for CHIP-like proteins in the Bombyx mori databases, we found a novel silkworm gene, Bombyx mori CHIP. Phylogenetic analysis showed that BmCHIP belongs to Lepidopteran lineages. Quantitative reverse transcription-PCR analysis indicated that BmCHIP was relatively highly expressed in the gonad and fat body. A pull-down experiment and auto-ubiquitination assay showed that BmCHIP interacted with BmHSC70 and had E3 ligase activity. Additionally, immunohistochemical analysis revealed that BmCHIP was partially co-localized with ubiquitin in BmN4 cells. These data support that BmCHIP plays an important role in the ubiquitin proteasome system as an E3 ubiquitin ligase in B. mori. Copyright © 2016 Elsevier Inc. All rights reserved.

  4. Conserved region at the COOH terminus of human immunodeficiency virus gp120 envelope protein contains an immunodominant epitope

    International Nuclear Information System (INIS)

    Palker, T.J.; Matthews, T.J.; Clark, M.E.

    1987-01-01

    A highly immunogenic epitope from a conserved COOH-terminal region of the human immunodeficiency virus (HIV) gp120 envelope protein has been identified with antisera from HIV-seropositive subjects and a synthetic peptide (SP-22) containing 15 amino acids from this region (Ala-Pro-Thr-Lys-Ala-Lys-Arg-Arg-Val-Val-Gln-Arg-Glu-Lys-Arg). Peptide SP-22 absorbed up to 100% of anti-gp120 antibody reactivity from select HIV + patient sera in immunoblot assays and up to 79% of serum anti-gp120 antibody reactivity in competition RIA. In RIA, 45% of HIV-seropositive subjects had antibodies that bound to peptide SP-22. Human anti-SP-22 antibodies that bound to and were eluted from an SP-22 affinity column reacted with gp120 in RIA and immunoblot assays but did not neutralize HIV or inhibit HIV-induced syncytium formation in vitro, even though these antibodies comprised 70% of all anti-gp120 antibodies in the test serum. In contrast, the remaining 30% of SP-22 nonreactive anti-gp120 antibodies did not react with gp120 in immunoblot assays but did react in RIA and neutralized HIV in vitro. Thus, ≅ 50% of HIV-seropositive patients make high titers of nonneutralizing antibodies to an immunodominant antigen on gp120 defined by SP-22. Moreover, the COOH terminus of gp120 contains the major antigen or antigens identified by human anti-gp120 antibodies in immunoblot assays

  5. The carboxyl terminus of human cytomegalovirus-encoded 7 transmembrane receptor US28 camouflages agonism by mediating constitutive endocytosis

    DEFF Research Database (Denmark)

    Waldhoer, Maria; Casarosa, Paola; Rosenkilde, Mette M

    2003-01-01

    are separable entities in this viral chemokine receptor. We generated chimeric and mutant US28 proteins that were altered in either their constitutive endocytic (US28 Delta 300, US28 Delta 317, US28-NK1-ctail, and US28-ORF74-ctail) or signaling properties (US28R129A). By using this series of mutants, we show...... further show that the constitutive endocytic property of US28 affects the action of its chemokine ligand fractalkine/CX3CL1 and show that in the absence of the US28 C terminus, fractalkine/CX3CL1 acts as an agonist on US28. This demonstrates for the first time that the endocytic properties of a 7TM......US28 is one of four 7 transmembrane (7TM) chemokine receptors encoded by human cytomegalovirus and has been shown to both signal and endocytose in a ligand-independent, constitutively active manner. Here we show that the constitutive activity and constitutive endocytosis properties of US28...

  6. Creating Interdisciplinarity within Monodisciplinary Structures

    DEFF Research Database (Denmark)

    Lindvig, Katrine

    discrepancies in the use of the term interdisciplinarity, which have repercussions for the practices and incentives of creating interdisciplinary education, research and collaboration. Overall, the thesis shows that interdisciplinary teaching and learning practices have to engage in a continuous balancing...

  7. Positive effects of creating mandalas

    OpenAIRE

    Toroš, Maja

    2016-01-01

    The purpose of the present master’s thesis is to examine the psychological effects of creating mandalas and to give recommendations for a better use of the mandalas in the practice of art assistance in order to help in this way others and oneself. Mandalas are sacred symbols, used in Hinduism and Buddhist religion practices as tools for meditation and concentration. They are geometrical representations of the creation of the cosmos and schemas of all that was created. Jung sat the foundations...

  8. Creating visual explanations improves learning.

    Science.gov (United States)

    Bobek, Eliza; Tversky, Barbara

    2016-01-01

    Many topics in science are notoriously difficult for students to learn. Mechanisms and processes outside student experience present particular challenges. While instruction typically involves visualizations, students usually explain in words. Because visual explanations can show parts and processes of complex systems directly, creating them should have benefits beyond creating verbal explanations. We compared learning from creating visual or verbal explanations for two STEM domains, a mechanical system (bicycle pump) and a chemical system (bonding). Both kinds of explanations were analyzed for content and learning assess by a post-test. For the mechanical system, creating a visual explanation increased understanding particularly for participants of low spatial ability. For the chemical system, creating both visual and verbal explanations improved learning without new teaching. Creating a visual explanation was superior and benefitted participants of both high and low spatial ability. Visual explanations often included crucial yet invisible features. The greater effectiveness of visual explanations appears attributable to the checks they provide for completeness and coherence as well as to their roles as platforms for inference. The benefits should generalize to other domains like the social sciences, history, and archeology where important information can be visualized. Together, the findings provide support for the use of learner-generated visual explanations as a powerful learning tool.

  9. Mutations in the catalytic core or the C-terminus of murine leukemia virus (MLV) integrase disrupt virion infectivity and exert diverse effects on reverse transcription

    International Nuclear Information System (INIS)

    Steinrigl, Adolf; Nosek, Dagmara; Ertl, Reinhard; Guenzburg, Walter H.; Salmons, Brian; Klein, Dieter

    2007-01-01

    Understanding of the structures and functions of the retroviral integrase (IN), a key enzyme in the viral replication cycle, is essential for developing antiretroviral treatments and facilitating the development of safer gene therapy vehicles. Thus, four MLV IN-mutants were constructed in the context of a retroviral vector system, harbouring either a substitution in the catalytic centre, deletions in the C-terminus, or combinations of both modifications. IN-mutants were tested for their performance in different stages of the viral replication cycle: RNA-packaging; RT-activity; transient and stable infection efficiency; dynamics of reverse transcription and nuclear entry. All mutant vectors packaged viral RNA with wild-type efficiencies and displayed only slight reductions in RT-activity. Deletion of either the IN C-terminus alone, or in addition to part of the catalytic domain exerted contrasting effects on intracellular viral DNA levels, implying that IN influences reverse transcription in more than one direction

  10. Role of the N-terminus for the stability of an amyloid-β fibril with three-fold symmetry.

    Directory of Open Access Journals (Sweden)

    Christian A Söldner

    Full Text Available A key player in Alzheimer's disease is the peptide amyloid-beta (Aβ, whose aggregation into small soluble oligomers, protofilaments, and fibrils finally leads to plaque deposits in human brains. The aggregation behavior of Aβ is strongly modulated by the nature and composition of the peptide's environment and by its primary sequence properties. The N-terminal residues of Aβ play an important role, because they are known to change the peptide's aggregation propensity. Since these residues are for the first time completely resolved at the molecular level in a three-fold symmetric fibril structure derived from a patient, we chose that system as template for a systematic investigation of the influence of the N-terminus upon structural stability. Using atomistic molecular dynamics simulations, we examined several fibrillar systems comprising three, six, twelve and an infinite number of layers, both with and without the first eight residues. First, we found that three layers are not sufficient to stabilize the respective Aβ topology. Second, we observed a clear stabilizing effect of the N-terminal residues upon the overall fibril fold: truncated Aβ systems were less stable than their full-length counterparts. The N-terminal residues Arg5, Asp7, and Ser8 were found to form important interfilament contacts stabilizing the overall fibril structure of three-fold symmetry. Finally, similar structural rearrangements of the truncated Aβ species in different simulations prompted us to suggest a potential mechanism involved in the formation of amyloid fibrils with three-fold symmetry.

  11. The functional domain of GCS1-based gamete fusion resides in the amino terminus in plant and parasite species.

    Directory of Open Access Journals (Sweden)

    Toshiyuki Mori

    Full Text Available Fertilization is one of the most important processes in all organisms utilizing sexual reproduction. In a previous study, we succeeded in identifying a novel male gametic transmembrane protein GCS1 (GENERATIVE CELL SPECIFIC 1, also called HAP2 (HAPLESS 2 in the male-sterile Arabidopsis thaliana mutants, as a factor critical to gamete fusion in flowering plants. Interestingly, GCS1 is highly conserved among various eukaryotes covering plants, protists and invertebrates. Of these organisms, Chlamydomonas (green alga and Plasmodium (malaria parasite GCS1s similarly show male gametic expression and gamete fusion function. Since it is generally believed that protein factors controlling gamete fusion have rapidly evolved and different organisms utilize species-specific gamete fusion factors, GCS1 may be an ancient fertilization factor derived from the common ancestor of those organisms above. And therefore, its molecular structure and function are important to understanding the common molecular mechanics of eukaryotic fertilization. In this study, we tried to detect the central functional domain(s of GCS1, using complementation assay of Arabidopsis GCS1 mutant lines expressing modified GCS1. As a result, the positively-charged C-terminal sequence of this protein is dispensable for gamete fusion, while the highly conserved N-terminal domain is critical to GCS1 function. In addition, in vitro fertilization assay of Plasmodium berghei (mouse malaria parasite knock-in lines expressing partly truncated GCS1 showed similar results. Those findings above indicate that the extracellular N-terminus alone is sufficient for GCS1-based gamete fusion.

  12. Identification of a heterologous cellulase and its N-terminus that can guide recombinant proteins out of Escherichia coli.

    Science.gov (United States)

    Gao, Dongfang; Wang, Shengjun; Li, Haoran; Yu, Huili; Qi, Qingsheng

    2015-04-10

    The Gram-negative bacterium Escherichia coli has been widely used as a cell factory for the production of proteins and specialty chemicals because it is the best characterized host with many available expression and regulation systems. However, recombinant proteins produced in Escherichia coli are generally intracellular and often found in the form of inclusion bodies. Extracellular production of proteins is advantageous compared with intracellular production because extracellular proteins can be purified more easily and can avoid protease attack, which results in higher product quality. In this study, we found a catalytic domain of a cellulase (Cel-CD) and its N-terminus can be employed as carriers for extracellular production of recombinant proteins. In this report, we identified the catalytic domain of a cellulase (Cel-CD) from Bacillus sp. that can be secreted into the medium from recombinant E. coli BL21 (DE3) in large quantities without its native signal peptide. By subcellular location analysis, we proved that the secretion was a two-step process and the N-terminal sequence of the full length Cel-CD played a crucial function in secretion. Both the Cel-CD and its N-terminal sequence can serve as carriers for efficient extracellular production of select target proteins. Fusion of heterologous proteins with N20 from Cel-CD can carry the target proteins out of the cells with a concentration from 101 to 691 mg/L in flask cultivation. The extracellular recombinant proteins with a relative high purity. The results suggested that this system has a potential application in plant biomass conversion and industrial production of enzymes and therapeutic proteins.

  13. Atomic Structure and Biochemical Characterization of an RNA Endonuclease in the N Terminus of Andes Virus L Protein.

    Directory of Open Access Journals (Sweden)

    Yaiza Fernández-García

    2016-06-01

    Full Text Available Andes virus (ANDV is a human-pathogenic hantavirus. Hantaviruses presumably initiate their mRNA synthesis by using cap structures derived from host cell mRNAs, a mechanism called cap-snatching. A signature for a cap-snatching endonuclease is present in the N terminus of hantavirus L proteins. In this study, we aimed to solve the atomic structure of the ANDV endonuclease and characterize its biochemical features. However, the wild-type protein was refractory to expression in Escherichia coli, presumably due to toxic enzyme activity. To circumvent this problem, we introduced attenuating mutations in the domain that were previously shown to enhance L protein expression in mammalian cells. Using this approach, 13 mutant proteins encompassing ANDV L protein residues 1-200 were successfully expressed and purified. Protein stability and nuclease activity of the mutants was analyzed and the crystal structure of one mutant was solved to a resolution of 2.4 Å. Shape in solution was determined by small angle X-ray scattering. The ANDV endonuclease showed structural similarities to related enzymes of orthobunya-, arena-, and orthomyxoviruses, but also differences such as elongated shape and positively charged patches surrounding the active site. The enzyme was dependent on manganese, which is bound to the active site, most efficiently cleaved single-stranded RNA substrates, did not cleave DNA, and could be inhibited by known endonuclease inhibitors. The atomic structure in conjunction with stability and activity data for the 13 mutant enzymes facilitated inference of structure-function relationships in the protein. In conclusion, we solved the structure of a hantavirus cap-snatching endonuclease, elucidated its catalytic properties, and present a highly active mutant form, which allows for inhibitor screening.

  14. The nature of the Syntaxin4 C-terminus affects Munc18c-supported SNARE assembly.

    Directory of Open Access Journals (Sweden)

    Asma Rehman

    Full Text Available Vesicular transport of cellular cargo requires targeted membrane fusion and formation of a SNARE protein complex that draws the two apposing fusing membranes together. Insulin-regulated delivery and fusion of glucose transporter-4 storage vesicles at the cell surface is dependent on two key proteins: the SNARE integral membrane protein Syntaxin4 (Sx4 and the soluble regulatory protein Munc18c. Many reported in vitro studies of Munc18c:Sx4 interactions and of SNARE complex formation have used soluble Sx4 constructs lacking the native transmembrane domain. As a consequence, the importance of the Sx4 C-terminal anchor remains poorly understood. Here we show that soluble C-terminally truncated Sx4 dissociates more rapidly from Munc18c than Sx4 where the C-terminal transmembrane domain is replaced with a T4-lysozyme fusion. We also show that Munc18c appears to inhibit SNARE complex formation when soluble C-terminally truncated Sx4 is used but does not inhibit SNARE complex formation when Sx4 is C-terminally anchored (by a C-terminal His-tag bound to resin, by a C-terminal T4L fusion or by the native C-terminal transmembrane domain in detergent micelles. We conclude that the C-terminus of Sx4 is critical for its interaction with Munc18c, and that the reported inhibitory role of Munc18c may be an artifact of experimental design. These results support the notion that a primary role of Munc18c is to support SNARE complex formation and membrane fusion.

  15. Insight into the Structural and Biological Relevance of the T/R Transition of the N-Terminus of the B-Chain in Human Insulin

    Czech Academy of Sciences Publication Activity Database

    Kosinová, Lucie; Veverka, Václav; Novotná, P.; Collinsová, Michaela; Urbanová, M.; Moody, N. R.; Turkenburg, J. P.; Jiráček, Jiří; Brzozowski, A. M.; Žáková, Lenka

    2014-01-01

    Roč. 53, č. 21 (2014), s. 3392-3402 ISSN 0006-2960 R&D Projects: GA ČR GPP207/11/P430; GA ČR GAP208/11/0105; GA MŠk(CZ) LK11205 Institutional support: RVO:61388963 Keywords : insulin * structure * N-terminus * B-chain * T/R transition Subject RIV: CE - Biochemistry Impact factor: 3.015, year: 2014

  16. C-terminus of the P4-ATPase ATP8A2 functions in protein folding and regulation of phospholipid flippase activity.

    Science.gov (United States)

    Chalat, Madhavan; Moleschi, Kody; Molday, Robert S

    2017-02-01

    ATP8A2 is a P4-ATPase that flips phosphatidylserine and phosphatidylethanolamine across cell membranes. This generates membrane phospholipid asymmetry, a property important in many cellular processes, including vesicle trafficking. ATP8A2 deficiency causes severe neurodegenerative diseases. We investigated the role of the C-terminus of ATP8A2 in its expression, subcellular localization, interaction with its subunit CDC50A, and function as a phosphatidylserine flippase. C-terminal deletion mutants exhibited a reduced tendency to solubilize in mild detergent and exit the endoplasmic reticulum. The solubilized protein, however, assembled with CDC50A and displayed phosphatidylserine flippase activity. Deletion of the C-terminal 33 residues resulted in reduced phosphatidylserine-dependent ATPase activity, phosphatidylserine flippase activity, and neurite extension in PC12 cells. These reduced activities were reversed with 60- and 80-residue C-terminal deletions. Unlike the yeast P4-ATPase Drs2, ATP8A2 is not regulated by phosphoinositides but undergoes phosphorylation on the serine residue within a CaMKII target motif. We propose a model in which the C-terminus of ATP8A2 consists of an autoinhibitor domain upstream of the C-terminal 33 residues and an anti-autoinhibitor domain at the extreme C-terminus. The latter blocks the inhibitory activity of the autoinhibitor domain. We conclude that the C-terminus plays an important role in the efficient folding and regulation of ATP8A2. © 2017 Chalat et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

  17. Surface targeting of the dopamine transporter involves discrete epitopes in the distal C terminus but does not require canonical PDZ domain interactions.

    Science.gov (United States)

    Bjerggaard, Christian; Fog, Jacob U; Hastrup, Hanne; Madsen, Kenneth; Loland, Claus J; Javitch, Jonathan A; Gether, Ulrik

    2004-08-04

    The human dopamine transporter (hDAT) contains a C-terminal type 2 PDZ (postsynaptic density 95/Discs large/zona occludens 1) domain-binding motif (LKV) known to interact with PDZ domain proteins such as PICK1 (protein interacting with C-kinase 1). As reported previously, we found that, after deletion of this motif, hDAT was retained in the endoplasmic reticulum (ER) of human embryonic kidney (HEK) 293 and Neuro2A cells, suggesting that PDZ domain interactions might be critical for hDAT targeting. Nonetheless, substitution of LKV with SLL, the type 1 PDZ-binding sequence from the beta2-adrenergic receptor, did not disrupt plasma membrane targeting. Moreover, the addition of an alanine to the hDAT C terminus (+Ala), resulting in an LKVA termination sequence, or substitution of LKV with alanines (3xAla_618-620) prevented neither plasma membrane targeting nor targeting into sprouting neurites of differentiated N2A cells. The inability of +Ala and 3xAla_618-620 to bind PDZ domains was confirmed by lack of colocalization with PICK1 in cotransfected HEK293 cells and by the inability of corresponding C-terminal fusion proteins to pull down purified PICK1. Thus, although residues in the hDAT C terminus are indispensable for proper targeting, PDZ domain interactions are not required. By progressive substitutions with beta2-adrenergic receptor sequence, and by triple-alanine substitutions in the hDAT C terminus, we examined the importance of epitopes preceding the LKV motif. Substitution of RHW(615-617) with alanines caused retention of the transporter in the ER despite preserved ability of this mutant to bind PICK1. We propose dual roles of the hDAT C terminus: a role independent of PDZ interactions for ER export and surface targeting, and a not fully clarified role involving PDZ interactions with proteins such as PICK1.

  18. Interaction of a peptide derived from C-terminus of human TRPA1 channel with model membranes mimicking the inner leaflet of the plasma membrane.

    Science.gov (United States)

    Witschas, Katja; Jobin, Marie-Lise; Korkut, Dursun Nizam; Vladan, Maria Magdalena; Salgado, Gilmar; Lecomte, Sophie; Vlachova, Viktorie; Alves, Isabel D

    2015-05-01

    The transient receptor potential ankyrin 1 channel (TRPA1) belongs to the TRP cation channel superfamily that responds to a panoply of stimuli such as changes in temperature, calcium levels, reactive oxygen and nitrogen species and lipid mediators among others. The TRP superfamily has been implicated in diverse pathological states including neurodegenerative disorders, kidney diseases, inflammation, pain and cancer. The intracellular C-terminus is an important regulator of TRP channel activity. Studies with this and other TRP superfamily members have shown that the C-terminus association with lipid bilayer alters channel sensitivity and activation, especially interactions occurring through basic residues. Nevertheless, it is not yet clear how this process takes place and which regions in the C-terminus would be responsible for such membrane recognition. With that in mind, herein the first putative membrane interacting region of the C-terminus of human TRPA1, (corresponding to a 29 residue peptide, IAEVQKHASLKRIAMQVELHTSLEKKLPL) named H1 due to its potential helical character was chosen for studies of membrane interaction. The affinity of H1 to lipid membranes, H1 structural changes occurring upon this interaction as well as effects of this interaction in lipid organization and integrity were investigated using a biophysical approach. Lipid models systems composed of zwitterionic and anionic lipids, namely those present in the lipid membrane inner leaflet, where H1 is prone to interact, where used. The study reveals a strong interaction and affinity of H1 as well as peptide structuration especially with membranes containing anionic lipids. Moreover, the interactions and peptide structure adoption are headgroup specific. Copyright © 2015 Elsevier B.V. All rights reserved.

  19. Cathepsin C and plasma glutamate carboxypeptidase secreted from Fischer rat thyroid cells liberate thyroxin from the N-terminus of thyroglobulin.

    Science.gov (United States)

    Suban, Dejan; Zajc, Tajana; Renko, Miha; Turk, Boris; Turk, Vito; Dolenc, Iztok

    2012-03-01

    The release of a thyroid hormone from thyroglobulin is controlled by a complex regulatory system. We focused on the extracellular action of two lysosomal enzymes, cathepsin C (catC, dipeptidyl peptidase I) and PGCP (lysosomal dipeptidase), on thyroglobulin, and their ability to liberate the hormone thyroxin. Cathepsin C, an exopeptidase, removes dipeptides from the N-terminus of substrates, and PGCP hydrolyses dipeptides to amino acids. In vitro experiments proved that cathepsin C removes up to 12 amino acids from the N-terminus of porcine thyroglobulin, including a dipeptide with thyroxin on position 5. The newly formed N-terminus, Arg-Pro-, was not hydrolysed further by cathepsin C. Cell culture experiments with FRTL-5 cell line showed localization of cathepsin C and PGCP and their secretion into the medium. Secretion of the active cathepsin C from FRTL-5 cells is stimulated by TSH, insulin, and/or somatostatin. The released enzymes liberate thyroxin from porcine thyroglobulin added to media. The hormone liberation can be reduced by synthetic inhibitors of cysteine proteinases and metalloproteinases. Additionally, we show that TSH, insulin, and/or somatostatin induce up-regulation of N-acetylglucosaminyltransferase 1, the enzyme responsible for the initiation of biosynthesis of hybrid and complex N-glycosylation of proteins. Copyright © 2011 Elsevier Masson SAS. All rights reserved.

  20. Conserved amino acids within the N-terminus of the West Nile virus NS4A protein contribute to virus replication, protein stability and membrane proliferation

    International Nuclear Information System (INIS)

    Ambrose, R.L.; Mackenzie, J.M.

    2015-01-01

    The West Nile virus strain Kunjin virus (WNV KUN ) NS4A protein is a multifunctional protein involved in many aspects of the virus life-cycle and is a major component of the WNV KUN replication complex (RC). Previously we identified a conserved region in the C-terminus of NS4A regulating proteolytic processing and RC assembly, and now investigate key conserved residues in the N-terminus of NS4A and their contribution to WNV KUN replication. Mutation of P13 completely ablated replication, whereas, mutation of P48 and D49, near the first transmembrane helix, and G66 within the helix, showed variable defects in replication, virion secretion and membrane proliferation. Intriguingly, the P48 and G66 NS4A mutants resulted in specific proteasome depletion of NS4A that could in part be rescued with a proteasome inhibitor. Our results suggest that the N-terminus of NS4A contributes to correct folding and stability, essential for facilitating the essential roles of NS4A during replication. - Highlights: • Mutation of Proline13 of the WNV NS4A protein is lethal to replication. • 1st TMB helix of NS4A contributes to protein stability and membrane remodelling. • Unstable mutants of NS4A can be rescued with a proteasome inhibitor. • This study (and of others) contributes to a functional mapping of the NS4A protein

  1. On Creating and Sustaining Alternatives

    DEFF Research Database (Denmark)

    Kyng, Morten

    2015-01-01

    This paper presents and discusses an initiative aimed at creating direct and long lasting influence on the use and development of telemedicine and telehealth by healthcare professionals, patients and citizens. The initiative draws on ideas, insights, and lessons learned from Participatory Design...... (PD) as well as from innovation theory and software ecosystems. Last, but not least, the ongoing debate on public finances/economy versus tax evasion by major private companies has been an important element in shaping the vision and creating support for the initiative. This vision is about democratic...

  2. Journaling: creating space for "I".

    Science.gov (United States)

    Charles, Jennell P

    2010-01-01

    As nurses engaged in a caring profession, it is critical that we learn not only to care for others but also to care for ourselves. To care effectively for ourselves, we must create the space and time in which to do this. Journaling is one tool that scholars offer as a way to create this space. Although there is no clear consensus about the best techniques for journaling, there is evidence that journaling, as a reflective, meditative activity, can promote creativity, self-awareness, and personal development.

  3. Intentionality, consciousness, and creating community.

    Science.gov (United States)

    Malinski, Violet M

    2009-01-01

    Intentionality is briefly explored from the perspective of seminal written works on therapeutic touch and recorded conversations with Martha E. Rogers. This overview hints at possible interrelationships among intentionality, consciousness, and creating community, along with conceptual ambiguities, which are explored in detail by Zahourek and Larkin in this column.

  4. We create our own reality

    CERN Multimedia

    2003-01-01

    " Yes, we create our own reality. This is one of the most fundamental tenets of the ancient oriental religions, such as Buddhism. And during the last century, modern particle physics or quantum mechanics has discovered exactly the same thing" (1 page).

  5. Creating Space for Children's Literature

    Science.gov (United States)

    Serafini, Frank

    2011-01-01

    As teachers struggle to balance the needs of their students with the requirements of commercial reading materials, educators need to consider how teachers will create space for children's literature in today's classrooms. In this article, 10 practical recommendations for incorporating children's literature in the reading instructional framework…

  6. Creating an Innovative Learning Organization

    Science.gov (United States)

    Salisbury, Mark

    2010-01-01

    This article describes how to create an innovative learning (iLearning) organization. It begins by discussing the life cycle of knowledge in an organization, followed by a description of the theoretical foundation for iLearning. Next, the article presents an example of iLearning, followed by a description of the distributed nature of work, the…

  7. Creating a New Teaching Profession

    Science.gov (United States)

    Goldhaber, Dan, Ed.; Hannaway, Jane, Ed.

    2009-01-01

    Considering that having a quality teacher is the foremost in-school predictor of students' success, ensuring teacher excellence is vital to the nation's educational system. In "Creating a New Teaching Profession," diverse scholars assess the state of human capital development in the teaching profession today and how to progress.

  8. Gαq protein carboxyl terminus imitation polypeptide GCIP-27 improves cardiac function in chronic heart failure rats.

    Directory of Open Access Journals (Sweden)

    Xiao Lan Lu

    Full Text Available Gαq protein carboxyl terminus imitation polypeptide (GCIP-27 has been shown to alleviate pathological cardiomyocyte hypertrophy induced by various factors. Pathological cardiac hypertrophy increases the morbidity and mortality of cardiovascular diseases while it compensates for poor heart function. This study was designed to investigate the effects of GCIP-27 on heart function in rats with heart failure induced by doxorubicin.Forty-eight rats were randomly divided into the following six groups receiving vehicle (control, doxorubicin (Dox, losartan (6 mg/kg, i.g. and three doses of GCIP-27 (10, 30, 90 μg/kg; i.p., bid, respectively. Heart failure was induced by Dox, which was administered at a 20 mg/kg cumulative dose. After 10 weeks of treatment, we observed that GCIP-27 (30, 90 μg/kg significantly increased ejection fraction, fraction shortening, stroke volume and sarcoplasmic reticulum Ca2+ ATPase activity of Dox-treated hearts. Additionally, GCIP-27 decreased myocardial injury, heart weight index and left ventricular weight index, fibrosis and serum cardiac troponin-I concentration in Dox-treated mice. Immunohistochemistry, western blotting and real-time PCR experiments indicated that GCIP-27 (10-90 μg/kg could markedly upregulate the protein expression of myocardial α-myosin heavy chain (MHC, Bcl-2, protein kinase C (PKC ε and phosphorylated extracellular signal-regulated kinase (p-ERK 1/2 as well as the mRNA expression of α-MHC, but downregulated the expression of β-MHC, Bax and PKC βII, and the mRNA expression levels of β-MHC in Dox-treated mice. It was also found that GCIP-27 (30, 90 μg/L decreased cell size and protein content of cardiomyocytes significantly in vitro by comparison of Dox group.GCIP-27 could effectively ameliorate heart failure development induced by Dox. PKC-ERK1/2 signaling might represent the underlying mechanism of the beneficial effects of GCIP-27.

  9. Prognostic implications of carboxyl-terminus of Hsc70 interacting protein and lysyl-oxidase expression in human breast cancer

    Directory of Open Access Journals (Sweden)

    Patani Neill

    2010-01-01

    Full Text Available Background: Ubiquitin modification of proteins influences cellular processes relevant to carcinogenesis. CHIP (carboxyl-terminus of Hsc70-interacting protein is a chaperone-dependent E3 ubiquitin ligase, regulating the stability of heat shock protein 90 (HSP90 interacting proteins. CHIP is implicated in the modulation of estrogen receptor (ESR1 and Her-2/neu (ERBB2 stability. LOX (lysyl-oxidase serves intracellular roles and catalyses the cross-linking of extracellular matrix (ECM collagens and elastin. LOX expression is altered in human malignancies and their peri-tumoral stroma. However, paradoxical roles are reported. In this study, the level of mRNA expression of CHIP and LOX were assessed in normal and malignant breast tissue and correlated with clinico-pathological parameters. Materials and Methods: Breast cancer (BC tissues (n = 127 and normal tissues (n = 33 underwent RNA extraction and reverse transcription; transcript levels were determined using real-time quantitative PCR and normalized against CK-19. Transcript levels were analyzed against TNM stage, nodal involvement, tumor grade and clinical outcome over a ten-year follow-up period. Results: CHIP expression decreased with increasing Nottingham Prognostic Index (NPI: NPI-1 vs. NPI-3 (12.2 vs. 0.2, P = 0.0264, NPI-2 vs. NPI-3 (3 vs. 0.2, P = 0.0275. CHIP expression decreased with increasing TNM stage: TNM-1 vs. TNM-2 (12 vs. 0, P = 0.0639, TNM-1 vs. TNM-2-4 (12 vs. 0, P = 0.0434. Lower transcript levels were associated with increasing tumor grade: grade 1 vs. grade 3 (17.7 vs. 0.3, P = 0.0266, grade 2 vs. grade 3 (5 vs. 0.3, P = 0.0454. The overall survival (OS for tumors classified as ′low-level expression′, was poorer than those with ′high-level expression′ (118.1 vs. 152.3 months, P = 0.039. LOX expression decreased with increasing NPI: NPI-1 vs. NPI-2 (3 vs. 0, P = 0.0301 and TNM stage: TNM-1 = 3854639, TNM-2 = 908900, TNM-3 = 329, TNM-4 = 1.232 (P = NS. Conclusion: CHIP

  10. Leadership Networking Connect, Collaborate, Create

    CERN Document Server

    (CCL), Center for Creative Leadership; Baldwin, David

    2011-01-01

    Networking is essential to effective leadership in today's organizations. Leaders who are skilled networkers have access to people, information, and resources to help solve problems and create opportunities. Leaders who neglect their networks are missing out on a critical component of their role as leaders. This book will help leaders take a new view of networking and provide insight into how to enhance their networks and become effective at leadership networking.

  11. Creating a digital medical illustration.

    Science.gov (United States)

    Culley, Joanna

    2016-01-01

    This paper covers the steps required to complete a medical illustration in a digital format using Adobe Illustrator and Photoshop. The project example is the surgical procedure for the release of the glenohumeral joint for the condition known as 'frozen shoulder'. The purpose is to demonstrate one method which an artist can use within digital media to create a colour illustration such as the release of the glenohumeral joint. Included is a general overview as how to deal with the administration of a medical illustration commission through the experience of a professional freelance artist.

  12. Physical interaction between replication protein A (RPA) and MRN: involvement of RPA2 phosphorylation and the N-terminus of RPA1.

    Science.gov (United States)

    Oakley, Greg G; Tillison, Kristin; Opiyo, Stephen A; Glanzer, Jason G; Horn, Jeffrey M; Patrick, Steve M

    2009-08-11

    Replication protein A (RPA) is a heterotrimeric protein consisting of RPA1, RPA2, and RPA3 subunits that binds to single-stranded DNA (ssDNA) with high affinity. The response to replication stress requires the recruitment of RPA and the MRE11-RAD50-NBS1 (MRN) complex. RPA bound to ssDNA stabilizes stalled replication forks by recruiting checkpoint proteins involved in fork stabilization. MRN can bind DNA structures encountered at stalled or collapsed replication forks, such as ssDNA-double-stranded DNA (dsDNA) junctions or breaks, and promote the restart of DNA replication. Here, we demonstrate that RPA2 phosphorylation regulates the assembly of DNA damage-induced RPA and MRN foci. Using purified proteins, we observe a direct interaction between RPA with both NBS1 and MRE11. By utilizing RPA bound to ssDNA, we demonstrate that substituting RPA with phosphorylated RPA or a phosphomimetic weakens the interaction with the MRN complex. Also, the N-terminus of RPA1 is a critical component of the RPA-MRN protein-protein interaction. Deletion of the N-terminal oligonucleotide-oligosaccharide binding fold (OB-fold) of RPA1 abrogates interactions of RPA with MRN and individual proteins of the MRN complex. Further identification of residues critical for MRN binding in the N-terminus of RPA1 shows that substitution of Arg31 and Arg41 with alanines disrupts the RPA-MRN interaction and alters cell cycle progression in response to DNA damage. Thus, the N-terminus of RPA1 and phosphorylation of RPA2 regulate RPA-MRN interactions and are important in the response to DNA damage.

  13. c-Jun binds the N terminus of human TAF(II)250 to derepress RNA polymerase II transcription in vitro.

    Science.gov (United States)

    Lively, T N; Ferguson, H A; Galasinski, S K; Seto, A G; Goodrich, J A

    2001-07-06

    c-Jun is an oncoprotein that activates transcription of many genes involved in cell growth and proliferation. We studied the mechanism of transcriptional activation by human c-Jun in a human RNA polymerase II transcription system composed of highly purified recombinant and native transcription factors. Transcriptional activation by c-Jun depends on the TATA-binding protein (TBP)-associated factor (TAF) subunits of transcription factor IID (TFIID). Protein-protein interaction assays revealed that c-Jun binds with high specificity to the largest subunit of human TFIID, TAF(II)250. The region of TAF(II)250 bound by c-Jun lies in the N-terminal 163 amino acids. This same region of TAF(II)250 binds to TBP and represses its interaction with TATA boxes, thereby decreasing DNA binding by TFIID. We hypothesized that c-Jun is capable of derepressing the effect of the TAF(II)250 N terminus on TFIID-driven transcription. In support of this hypothesis, we found that c-Jun increased levels of TFIID-driven transcription in vitro when added at high concentrations to a DNA template lacking activator protein 1 (AP-1) sites. Moreover, c-Jun blocked the repression of TBP DNA binding caused by the N terminus of TAF(II)250. In addition to revealing a mechanism by which c-Jun activates transcription, our studies provide the first evidence that an activator can bind directly to the N terminus of TAF(II)250 to derepress RNA polymerase II transcription in vitro.

  14. The long N-terminus of the human monocarboxylate transporter 8 is a target of ubiquitin-dependent proteasomal degradation which regulates protein expression and oligomerization capacity.

    Science.gov (United States)

    Zwanziger, Denise; Schmidt, Mathias; Fischer, Jana; Kleinau, Gunnar; Braun, Doreen; Schweizer, Ulrich; Moeller, Lars Christian; Biebermann, Heike; Fuehrer, Dagmar

    2016-10-15

    Monocarboxylate transporter 8 (MCT8) equilibrates thyroid hormones between the extra- and the intracellular sides. MCT8 exists either with a short or a long N-terminus, but potential functional differences between both variants are yet not known. We, therefore, generated MCT8 constructs which are different in N-terminal length: MCT8(1-613), MCT8(25-613), MCT8(49-613) and MCT8(75-613). The M75G substitution prevents translation of MCT8(75-613) and ensures expression of full-length MCT8 protein. The K56G substitution was made to prevent ubiquitinylation. Cell-surface expression, localization and proteasomal degradation were investigated using C-terminally GFP-tagged MCT8 constructs (HEK293 and MDCK1 cells) and oligomerization capacity was determined using N-terminally HA- and C-terminally FLAG-tagged MCT8 constructs (COS7 cells). MCT8(1-613)-GFP showed a lower protein expression than the shorter MCT8(75-613)-GFP protein. The proteasome inhibitor lactacystin increased MCT8(1-613)-GFP protein amount, suggesting proteasomal degradation of MCT8 with the long N-terminus. Ubiquitin conjugation of MCT8(1-613)-GFP was found by immuno-precipitation. A diminished ubiquitin conjugation caused by K56G substitution resulted in increased MCT8(1-613)-GFP protein expression. Sandwich ELISA was performed to investigate if the bands at higher molecular weight observed in Western blot analysis are due to MCT8 oligomerization, which was indeed shown. Our data imply a role of the long N-terminus of MCT8 as target of ubiquitin-dependent proteasomal degradation affecting MCT8 amount and subsequently oligomerization capacity. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

  15. Molecular characterization of partial fusion gene and C-terminus extension length of haemagglutinin-neuraminidase gene of recently isolated Newcastle disease virus isolates in Malaysia

    Directory of Open Access Journals (Sweden)

    Berhanu Ayalew

    2010-08-01

    Full Text Available Abstract Background Newcastle disease (ND, caused by Newcastle disease virus (NDV, is a highly contagious disease of birds and has been one of the major causes of economic losses in the poultry industry. Despite routine vaccination programs, sporadic cases have occasionally occurred in the country and remain a constant threat to commercial poultry. Hence, the present study was aimed to characterize NDV isolates obtained from clinical cases in various locations of Malaysia between 2004 and 2007 based on sequence and phylogenetic analysis of partial F gene and C-terminus extension length of HN gene. Results The coding region of eleven NDV isolates fusion (F gene and carboxyl terminal region of haemagglutinin-neuraminidase (HN gene including extensions were amplified by reverse transcriptase PCR and directly sequenced. All the isolates have shown to have non-synonymous to synonymous base substitution rate ranging between 0.081 - 0.264 demonstrating presence of negative selection. Analysis based on F gene showed the characterized isolates possess three different types of protease cleavage site motifs; namely 112RRQKRF117, 112RRRKRF117 and 112GRQGRL117 and appear to show maximum identities with isolates in the region such as cockatoo/14698/90 (Indonesia, Ch/2000 (China, local isolate AF2240 indicating the high similarity of isolates circulating in the South East Asian countries. Meanwhile, one of the isolates resembles commonly used lentogenic vaccine strains. On further characterization of the HN gene, Malaysian isolates had C-terminus extensions of 0, 6 and 11 amino acids. Analysis of the phylogenetic tree revealed that the existence of three genetic groups; namely, genotype II, VII and VIII. Conclusions The study concluded that the occurrence of three types of NDV genotypes and presence of varied carboxyl terminus extension lengths among Malaysian isolates incriminated for sporadic cases.

  16. Molecular characterization of partial fusion gene and C-terminus extension length of haemagglutinin-neuraminidase gene of recently isolated Newcastle disease virus isolates in Malaysia.

    Science.gov (United States)

    Berhanu, Ayalew; Ideris, Aini; Omar, Abdul R; Bejo, Mohd Hair

    2010-08-08

    Newcastle disease (ND), caused by Newcastle disease virus (NDV), is a highly contagious disease of birds and has been one of the major causes of economic losses in the poultry industry. Despite routine vaccination programs, sporadic cases have occasionally occurred in the country and remain a constant threat to commercial poultry. Hence, the present study was aimed to characterize NDV isolates obtained from clinical cases in various locations of Malaysia between 2004 and 2007 based on sequence and phylogenetic analysis of partial F gene and C-terminus extension length of HN gene. The coding region of eleven NDV isolates fusion (F) gene and carboxyl terminal region of haemagglutinin-neuraminidase (HN) gene including extensions were amplified by reverse transcriptase PCR and directly sequenced. All the isolates have shown to have non-synonymous to synonymous base substitution rate ranging between 0.081 - 0.264 demonstrating presence of negative selection. Analysis based on F gene showed the characterized isolates possess three different types of protease cleavage site motifs; namely 112RRQKRF117, 112RRRKRF117 and 112GRQGRL117 and appear to show maximum identities with isolates in the region such as cockatoo/14698/90 (Indonesia), Ch/2000 (China), local isolate AF2240 indicating the high similarity of isolates circulating in the South East Asian countries. Meanwhile, one of the isolates resembles commonly used lentogenic vaccine strains. On further characterization of the HN gene, Malaysian isolates had C-terminus extensions of 0, 6 and 11 amino acids. Analysis of the phylogenetic tree revealed that the existence of three genetic groups; namely, genotype II, VII and VIII. The study concluded that the occurrence of three types of NDV genotypes and presence of varied carboxyl terminus extension lengths among Malaysian isolates incriminated for sporadic cases.

  17. Creating Cross-disciplinary Courses.

    Science.gov (United States)

    Reynolds, Elaine R

    2012-01-01

    Because of its focus on the biological underpinnings of action and behavior, neuroscience intersects with many fields of human endeavor. Some of these cross-disciplinary intersections have been long standing, while others, such as neurotheology or neuroeconomics, are more recently formed fields. Many undergraduate institutions have sought to include cross-disciplinary courses in their curriculum because this style of pedagogy is often seen as applicable to real world problems. However, it can be difficult for faculty with specialized training within their discipline to expand beyond their own fields to offer cross-disciplinary courses. I have been creating a series of multi- or cross-disciplinary courses and have found some strategies that have helped me successfully teach these classes. I will discuss general strategies and tools in developing these types of courses including: 1) creating mixed experience classrooms of students and contributing faculty 2) finding the right tools that will allow you to teach to a mixed population without prerequisites 3) examining the topic using multiple disciplinary perspectives 4) feeding off student experience and interest 5) assessing the impact of these courses on student outcomes and your neuroscience program. This last tool in particular is important in establishing the validity of this type of teaching for neuroscience students and the general student population.

  18. Creating your own leadership brand.

    Science.gov (United States)

    Kerfoot, Karlene

    2002-01-01

    Building equity in a brand happens through many encounters. The initial attraction must be followed by the meeting of expectations. This creates a loyalty that is part of an emotional connection to that brand. This is the same process people go through when they first meet a leader and decide if this is a person they want to buy into. People will examine your style, your competence, and your standards. If you fail on any of these fronts, your ability to lead will be severely compromised. People expect more of leaders now, because they know and recognize good leaders. And, predictably, people are now more cynical of leaders because of the well-publicized excess of a few leaders who advanced their own causes at the expense of their people and their financial future. This will turn out to be a good thing, because it will create a higher standard of leadership that all must aspire to achieve. When the bar is raised for us, our standards of performance are also raised.

  19. Creating nanoscale emulsions using condensation.

    Science.gov (United States)

    Guha, Ingrid F; Anand, Sushant; Varanasi, Kripa K

    2017-11-08

    Nanoscale emulsions are essential components in numerous products, ranging from processed foods to novel drug delivery systems. Existing emulsification methods rely either on the breakup of larger droplets or solvent exchange/inversion. Here we report a simple, scalable method of creating nanoscale water-in-oil emulsions by condensing water vapor onto a subcooled oil-surfactant solution. Our technique enables a bottom-up approach to forming small-scale emulsions. Nanoscale water droplets nucleate at the oil/air interface and spontaneously disperse within the oil, due to the spreading dynamics of oil on water. Oil-soluble surfactants stabilize the resulting emulsions. We find that the oil-surfactant concentration controls the spreading behavior of oil on water, as well as the peak size, polydispersity, and stability of the resulting emulsions. Using condensation, we form emulsions with peak radii around 100 nm and polydispersities around 10%. This emulsion formation technique may open different routes to creating emulsions, colloidal systems, and emulsion-based materials.

  20. OPPOSITIONS CREATING HOMOUR IN JOKES

    Directory of Open Access Journals (Sweden)

    Umral Deveci

    2016-12-01

    Full Text Available Human beings, who perceive the reality of death however who do not know when it will happen, begin their life with this deficiency. Therefore, throughout their lives, they struggle to consummate and make up for the things that they perceive as deficiency or shortcomings through different ways. Humor is one of these means. The fact that deficiencies are eliminated results in superiority and relaxation. The sense of humor and relaxation simultaneously provide laughter. When theories of humor such as superiority, incongruous and relief are taken into consideration, it seems that these theories are related and support each other. Each text is whole with its form and content, which should be evaluated as a whole as much as possible. Hence this study dwells on shortcomings in jokes and in the lights of these shortcomings and theories of humor, it is intended tomake humor in stories, in terms of structural and semantic context, more concrete. Five stories/jokes randomly selected through samples are analyzed in this article. There are two basic types of opposition. The firstone is opposition that creates situation, the second one is thatcreates laughter. The first opposition depicts the shortcomings of knowledge, skill, patience arrogance and jealousyand prepares the second opposition. The opposition that creates laughter make up for shortcomings through cause and effect relationship and laughter comes out.

  1. Creating an effective poster presentation.

    Science.gov (United States)

    Taggart, H M; Arslanian, C

    2000-01-01

    One way to build knowledge in nursing is to share research findings or clinical program outcomes. The dissemination of these findings is often a difficult final step in a project that has taken months or years to complete. One method of sharing findings in a relaxed and informal setting is a poster presentation. This method is an effective form for presenting findings using an interactive approach. The milieu of a poster presentation enables the presenters to interact and dialogue with colleagues. Guidelines for size and format require that the poster is clear and informative. Application of design helps to create visually appealing posters. This article summarizes elements of designing and conducting a poster presentation.

  2. Supporting SMEs in creating jobs

    Directory of Open Access Journals (Sweden)

    Daniela PASNICU

    2018-03-01

    Full Text Available SMEs are the most dynamic sector of the economy, representing a key factor in employment growth. Although SMEs have been severely affected by the economic crisis from 2008, they continue to be a more and more important source of generating jobs, regardless of size class. Given the differentiated contribution of SMEs to job creation as a result of their heterogeneity, the article analyses the dynamics of staff in enterprises by size classes and activities of the national economy. The statistical analysis is followed by a SWOT analysis of policies and tools to support SMEs in creating jobs both with direct action - management, recruitment, training and indirectly action by addressing general business issues. Measures that support the economic growth of SMEs through internationalization, innovation and improved access to finance foster competitiveness and, implicitly, the creation of new jobs.

  3. Creating breakthroughs at 3M.

    Science.gov (United States)

    von Hippel, E; Thomke, S; Sonnack, M

    1999-01-01

    Most senior managers want their product development teams to create break-throughs--new products that will allow their companies to grow rapidly and maintain high margins. But more often they get incremental improvements to existing products. That's partly because companies must compete in the short term. Searching for breakthroughs is expensive and time consuming; line extensions can help the bottom line immediately. In addition, developers simply don't know how to achieve breakthroughs, and there is usually no system in place to guide them. By the mid-1990s, the lack of such a system was a problem even for an innovative company like 3M. Then a project team in 3M's Medical-Surgical Markets Division became acquainted with a method for developing breakthrough products: the lead user process. The process is based on the fact that many commercially important products are initially thought of and even prototyped by "lead users"--companies, organizations, or individuals that are well ahead of market trends. Their needs are so far beyond those of the average user that lead users create innovations on their own that may later contribute to commercially attractive breakthroughs. The lead user process transforms the job of inventing breakthroughs into a systematic task of identifying lead users and learning from them. The authors explain the process and how the 3M project team successfully navigated through it. In the end, the team proposed three major new product lines and a change in the division's strategy that has led to the development of breakthrough products. And now several more divisions are using the process to break away from incrementalism.

  4. A potyvirus vector efficiently targets recombinant proteins to chloroplasts, mitochondria and nuclei in plant cells when expressed at the amino terminus of the polyprotein.

    Science.gov (United States)

    Majer, Eszter; Navarro, José-Antonio; Daròs, José-Antonio

    2015-09-01

    Plant virus-based expression systems allow quick and efficient production of recombinant proteins in plant biofactories. Among them, a system derived from tobacco etch virus (TEV; genus potyvirus) permits coexpression of equimolar amounts of several recombinant proteins. This work analyzed how to target recombinant proteins to different subcellular localizations in the plant cell using this system. We constructed TEV clones in which green fluorescent protein (GFP), with a chloroplast transit peptide (cTP), a nuclear localization signal (NLS) or a mitochondrial targeting peptide (mTP) was expressed either as the most amino-terminal product or embedded in the viral polyprotein. Results showed that cTP and mTP mediated efficient translocation of GFP to the corresponding organelle only when present at the amino terminus of the viral polyprotein. In contrast, the NLS worked efficiently at both positions. Viruses expressing GFP in the amino terminus of the viral polyprotein produced milder symptoms. Untagged GFPs and cTP and NLS tagged amino-terminal GFPs accumulated to higher amounts in infected tissues. Finally, viral progeny from clones with internal GFPs maintained the extra gene better. These observations will help in the design of potyvirus-based vectors able to coexpress several proteins while targeting different subcellular localizations, as required in plant metabolic engineering. Copyright © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  5. The importance of subfragment 2 and C-terminus of myosin heavy chain for thick filament assembly in skeletal muscle cells.

    Science.gov (United States)

    Ojima, Koichi; Oe, Mika; Nakajima, Ikuyo; Shibata, Masahiro; Muroya, Susumu; Chikuni, Koichi; Hattori, Akihito; Nishimura, Takanori

    2015-04-01

    In skeletal muscle cells, myofibrillar proteins are highly organized into sarcomeres in which thick filaments interdigitate with thin filaments to generate contractile force. The size of thick filaments, which consist mainly of myosin molecules, is strictly controlled. However, little is known about the mechanisms by which myosin molecules assemble into thick filaments. Here, we assessed the ability of each domain of myosin heavy chain (Myh) to form thick filaments. We showed that exogenously expressed subfragment 2 (S2) + light meromyosin (LMM) of Myh was efficiently incorporated into thick filaments in muscle cells, although neither solely expressed S2 nor LMM targeted to thick filaments properly. In nonmuscle COS7 cells, S2+LMM formed more enlarged filaments/speckles than LMM. These results suggest that Myh filament formation is induced by S2 accompanying LMM. We further examined the effects of Myh C-terminus on thick filament assembly. C-terminal deletion mutants were incorporated not into entire thick filaments but rather into restricted regions of thick filaments. Our findings suggest that the elongation of myosin filaments to form thick filaments is regulated by S2 as well as C-terminus of LMM. © 2014 Japanese Society of Animal Science.

  6. Bioenergetic Consequences of FLAG Tag Addition to the C-Terminus of Subunit 8 of Yeast Saccharomyces cerevisiae Mitochondrial ATP Synthase

    Directory of Open Access Journals (Sweden)

    I MADE ARTIKA

    2010-09-01

    Full Text Available The yeast mitochondrial F1F0-ATP synthase is a multisubunit complex that contains at least 17 different subunits. Subunit 8 of yeast mitochondrial ATP synthase is a hydrophobic protein of 48 amino acids encoded by the mitochondrial ATP8 gene. Subunit 8 has three distinct domains; an N-terminal domain, a central hydrophobic domain and a C-terminal domain. FLAG tag addition to subunit 8 protein potentially facilitate elucidation of its topology, structure, and function. It has been shown that following incorporation of FLAG tag to its C-terminus, subunit 8 still assemble into functional ATP synthase complex. In order to analyze bioenergetic consequences of the FLAG tag addition, a yeast strain expressing FLAG tagged-subunit 8 was subjected to cellular respiration assays. Results obtained showed that addition of FLAG tag to the C-terminus of subunit 8 does not impair its proper functioning. The FLAG tag system, therefore, can be employed to study subunit 8′s detailed structure, topology, and function.

  7. The carboxyl terminus of FANCE recruits FANCD2 to the Fanconi Anemia (FA) E3 ligase complex to promote the FA DNA repair pathway.

    Science.gov (United States)

    Polito, David; Cukras, Scott; Wang, Xiaozhe; Spence, Paige; Moreau, Lisa; D'Andrea, Alan D; Kee, Younghoon

    2014-03-07

    Fanconi anemia (FA) is a genome instability syndrome characterized by bone marrow failure and cellular hypersensitivity to DNA cross-linking agents. In response to DNA damage, the FA pathway is activated through the cooperation of 16 FA proteins. A central player in the pathway is a multisubunit E3 ubiquitin ligase complex or the FA core complex, which monoubiquitinates its substrates FANCD2 and FANCI. FANCE, a subunit of the FA core complex, plays an essential role by promoting the integrity of the complex and by directly recognizing FANCD2. To delineate its role in substrate ubiquitination from the core complex assembly, we analyzed a series of mutations within FANCE. We report that a phenylalanine located at the highly conserved extreme C terminus, referred to as Phe-522, is a critical residue for mediating the monoubiquitination of the FANCD2-FANCI complex. Using the FANCE mutant that specifically disrupts the FANCE-FANCD2 interaction as a tool, we found that the interaction-deficient mutant conferred cellular sensitivity in reconstituted FANCE-deficient cells to a similar degree as FANCE null cells, suggesting the significance of the FANCE-FANCD2 interaction in promoting cisplatin resistance. Intriguingly, ectopic expression of the FANCE C terminus fragment alone in FA normal cells disrupts DNA repair, consolidating the importance of the FANCE-FANCD2 interaction in the DNA cross-link repair.

  8. The Carboxyl Terminus of FANCE Recruits FANCD2 to the Fanconi Anemia (FA) E3 Ligase Complex to Promote the FA DNA Repair Pathway*

    Science.gov (United States)

    Polito, David; Cukras, Scott; Wang, Xiaozhe; Spence, Paige; Moreau, Lisa; D'Andrea, Alan D.; Kee, Younghoon

    2014-01-01

    Fanconi anemia (FA) is a genome instability syndrome characterized by bone marrow failure and cellular hypersensitivity to DNA cross-linking agents. In response to DNA damage, the FA pathway is activated through the cooperation of 16 FA proteins. A central player in the pathway is a multisubunit E3 ubiquitin ligase complex or the FA core complex, which monoubiquitinates its substrates FANCD2 and FANCI. FANCE, a subunit of the FA core complex, plays an essential role by promoting the integrity of the complex and by directly recognizing FANCD2. To delineate its role in substrate ubiquitination from the core complex assembly, we analyzed a series of mutations within FANCE. We report that a phenylalanine located at the highly conserved extreme C terminus, referred to as Phe-522, is a critical residue for mediating the monoubiquitination of the FANCD2-FANCI complex. Using the FANCE mutant that specifically disrupts the FANCE-FANCD2 interaction as a tool, we found that the interaction-deficient mutant conferred cellular sensitivity in reconstituted FANCE-deficient cells to a similar degree as FANCE null cells, suggesting the significance of the FANCE-FANCD2 interaction in promoting cisplatin resistance. Intriguingly, ectopic expression of the FANCE C terminus fragment alone in FA normal cells disrupts DNA repair, consolidating the importance of the FANCE-FANCD2 interaction in the DNA cross-link repair. PMID:24451376

  9. Creating responsible partnerships in tourism

    Directory of Open Access Journals (Sweden)

    Ana Spitzer

    2012-06-01

    Full Text Available RQ: Organisations do not provide sufficient time and effort to seek out companies for partners that would, with the assistance of responsible cooperation, contribute to better quality offers and consequently to increased income and the good reputation of both companies. Responsibilities and ethics is where organizations on bothsides would take on and accept their own norms, tasks, obligations and be aware that in a relationship there is a need to give explanations and justify one’s actions, such partnerships will be long and prosperous. This requires a great deal of knowledge and maturity together with a very important personal characteristic that is care. This study examines whether the creation of long term partnerships through responsible and more personal (friendlyrelations brings the organization to greater success.Purpose: The purpose of this research is to determine how important it is for organizations in the tourism industry to build long term relationships, what it should be based on and whether companies are willing to change the current methods of operations.Method: The method of research was an interview with individuals that had a certain position within a tourism company and had contacts with partners and were obligated to see out new ones. A paradigm model was built and the responses analysed.Results: The survey results are encouraging. The interviews showed that respondents were aware that it is necessary to have long term and responsible partnerships. They recognized that in today’s world there is a lack of collaboration that is based on understanding andthat there should be more relations on a personal level. It isrequired that this changes in the future. The participants specifically highlight financial irresponsibility in many companies that destroys collaboration.Organization: With the help of this study, the author attempts to contribute ideas to organizations on how to create solid collaboration with partners, as

  10. [Effect of manual acupuncture stimulation of "Zusanli" (ST 36) on gastric motility, and SP and motilin activities in gastric antrum and nucleus raphe magnus in gastric hyperactivity and hypoactivity rats].

    Science.gov (United States)

    Yan, Chun-Chuan; Peng, Yan; Lin, Ya-Ping; Yi, Shou-Xiang; Chen, Ping; Hou, Yan-Ling; Shi, Dong-Mei

    2013-10-01

    To observe the changes of gastric motility and levels of substance P (SP) and motilin (MTL) in the gastric antrum and Nucleus Raphe Magnus (NRM) after manual acupuncture stimulation of "Zusanli" (ST 36) in gastric hyperactivity and hypoactivity rats, so as to analyze the role of NRM in acupuncture mediated adjustment of gastric motility. Fifty SD rats were randomly and equally divided into control, gastric hyperactivity (G-Hypera) model, gastric hypoactivity (G-Hypoa) model, acupuncture + G-Hypera and acupuncture + G-Hypoa groups (10 rats/group). G-Hypera model was established by intravenous (tail vein) injection of Maxolon (0.5 mL/200 g) and G-Hypoa model established by intravenous injection of Atropin (0.5 mL/200 g), respectively. After insertion of acupuncture needles into bilateral "Zusanli" (ST 36), the needles were repeatedly manipulated at a frequency of about 2 Hz for 5 min. The intragastric pressure was recorded and analyzed using a physiological signal analysis system. The SP and MTL contents of gastric antrum were measured by ELISA, and SP and MTL immunoactivity of NRM was determined by immunohistochemistry. In gastric hyperactivity rats, compared with the control group, the intragastric pressure (not systolic frequency), SP and MTL contents in the gastric antrum and MTL immunoactivity of NRM were significantly increased (P effect on gastric motility, which is closely associated with its functions in regulating gastric SP and MTL level and the expression of MTL and SP in the NRM of brainstem.

  11. Creating experimental color harmony map

    Science.gov (United States)

    Chamaret, Christel; Urban, Fabrice; Lepinel, Josselin

    2014-02-01

    Starting in the 17th century with Newton, color harmony is a topic that did not reach a consensus on definition, representation or modeling so far. Previous work highlighted specific characteristics for color harmony on com- bination of color doublets or triplets by means of a human rating on a harmony scale. However, there were no investigation involving complex stimuli or pointing out how harmony is spatially located within a picture. The modeling of such concept as well as a reliable ground-truth would be of high value for the community, since the applications are wide and concern several communities: from psychology to computer graphics. We propose a protocol for creating color harmony maps from a controlled experiment. Through an eye-tracking protocol, we focus on the identification of disharmonious colors in pictures. The experiment was composed of a free viewing pass in order to let the observer be familiar with the content before a second pass where we asked "to search for the most disharmonious areas in the picture". Twenty-seven observers participated to the experiments that was composed of a total of 30 different stimuli. The high inter-observer agreement as well as a cross-validation confirm the validity of the proposed ground-truth.

  12. Creating a winning organizational culture.

    Science.gov (United States)

    Campbell, Robert James

    2009-01-01

    This article explores the idea of how to create a winning organizational culture. By definition, a winning organizational culture is one that is able to make current innovations stick, while continuously changing based on the demands of the marketplace. More importantly, the article explores the notion that a winning organizational culture can have a profound impact on the conscious of the workforce, helping each individual to become a better, more productive person, who provides important services and products to the community. To form a basis toward defining the structure of what a winning organization culture looks like, 4 experts were asked 12 questions related to the development of an organizational culture. Three of the experts have worked intimately within the health care industry, while a fourth has been charged with turning around an organization that has had a losing culture for 17 years. The article provides insight into the role that values, norms, goals, leadership style, familiarity, and hiring practices play in developing a winning organizational culture. The article also emphasizes the important role that leaders perform in developing an organizational culture.

  13. Role of cysteine residues in the carboxyl-terminus of the follicle-stimulating hormone receptor in intracellular traffic and postendocytic processing

    Directory of Open Access Journals (Sweden)

    Brenda Melo-Nava

    2016-07-01

    Full Text Available Posttranslational modifications occurring during the biosynthesis of G protein-coupled receptors include glycosylation and palmitoylation at conserved cysteine residues located in the carboxyl-terminus (Ctail of the receptor. In a number of these receptors, these modifications play an important role in receptor function and particularly, in intracellular trafficking. In the present study, the three cysteine residues present in the carboxyl-terminus of the human FSHR were replaced with glycine by site-directed mutagenesis. Wild-type and mutant (Cys627/629/655Gly FSHRs were then transiently expressed in HEK-293 cells and analyzed for cell-surface plasma membrane expression, agonist-stimulated signaling and internalization, and postendocytic processing in the abscence and presence of lysosome and/or proteasome inhibitors. Compared with the wild-type FSHR, the triple mutant FSHR exhibited ~70% reduction in plasma membrane expression as well as a profound attenuation in agonist-stimulated cAMP production and ERK1/2 phosphorylation. Incubation of HEK-293 cells expressing the wild-type FSHR with 2-bromopalmitate (palmitoylation inhibitor for 6 h, decreased plasma membrane expression of the receptor by ~30%. The internalization kinetics and β-arrestin 1 and 2 recruitment were similar between the wild-type and triple mutant FSHR as disclosed by assays performed in non-equilibrium binding conditions and by confocal microscopy. Cells expressing the mutant FSHR recycled the internalized FSHR back to the plasma membrane less efficiently than those expressing the wild-type FSHR, an effect that was counteracted by proteasome but not by lysosome inhibition. These results indicate that replacement of the cysteine residues present in the carboxyl-terminus of the FSHR, impairs receptor trafficking from the endoplasmic reticulum/Golgi apparatus to the plasma membrane and its recycling from endosomes back to the cell surface following agonist

  14. Implication of the C terminus of the Prunus necrotic ringspot virus movement protein in cell-to-cell transport and in its interaction with the coat protein.

    Science.gov (United States)

    Aparicio, Frederic; Pallás, Vicente; Sánchez-Navarro, Jesús

    2010-07-01

    The movement protein (MP) of Prunus necrotic ringspot virus (PNRSV) is required for viral transport. Previous analysis with MPs of other members of the family Bromoviridae has shown that the C-terminal part of these MPs plays a critical role in the interaction with the cognate coat protein (CP) and in cell-to-cell transport. Bimolecular fluorescence complementation and overlay analysis confirm an interaction between the C-terminal 38 aa of PNRSV MP and its cognate CP. Mutational analysis of the C-terminal region of the PNRSV MP revealed that its C-terminal 38 aa are dispensable for virus transport, however, the 4 aa preceding the dispensable C terminus are necessary to target the MP to the plasmodesmata and for the functionality of the protein. The capacity of the PNRSV MP to use either a CP-dependent or a CP-independent cell-to-cell transport is discussed.

  15. Structural Characterization of the Loop at the Alpha-Subunit C-Terminus of the Mixed Lineage Leukemia Protein Activating Protease Taspase1.

    Directory of Open Access Journals (Sweden)

    Johannes van den Boom

    Full Text Available Type 2 asparaginases, a subfamily of N-terminal nucleophile (Ntn hydrolases, are activated by limited proteolysis. This activation yields a heterodimer and a loop region at the C-terminus of the α-subunit is released. Since this region is unresolved in all type 2 asparaginase crystal structures but is close to the active site residues, we explored this loop region in six members of the type 2 asparaginase family using homology modeling. As the loop model for the childhood cancer-relevant protease Taspase1 differed from the other members, Taspase1 activation as well as the conformation and dynamics of the 56 amino acids loop were investigated by CD and NMR spectroscopy. We propose a helix-turn-helix motif, which can be exploited as novel anticancer target to inhibit Taspase1 proteolytic activity.

  16. The Energetics of Streptococcal Enolase Octamer Formation: The Quantitative Contributions of the Last Eight Amino Acids at the Carboxy-Terminus.

    Directory of Open Access Journals (Sweden)

    Jack A Kornblatt

    Full Text Available The enolase produced by Streptococcus pyogenes is a homo-octamer whose overall shape resembles that of a donut. The octamer is best described as a tetramer of dimers. As such, it contains two types of interfaces. The first is common to almost all enolases as most enolases that have been studied are dimers. The second is unique to the octamers and includes residues near the carboxy-terminus. The primary sequence of the enolase contains 435 residues with an added 19 as an N-terminal hexahistine tag. We have systematically truncated the carboxy-terminus, individually removing the first 8 residues. This gave rise to a series of eight structures containing respectively, 435, 434, 433, 432, 431, 430, 429 and 427 residues. The truncations cause the protein to gradually dissociate from octamers to enzymatically inactive monomers with very small amounts of intermediate tetramers and dimers. We have evaluated the contributions of the missing residues to the monomer/octamer equilibrium using a combination of analytical ultracentrifugation and activity assays. For the dissociation reaction, octamer 8 monomer truncation of all eight C-terminal residues resulted in a diminution in the standard Gibbs energy of dissociation of about 59 kJ/mole of octamer relative to the full length protein. Considering that this change is spread over eight subunits, this translates to a change in standard Gibbs interaction energy of less than 8 kJ/mole of monomer distributed over the eight monomers. The resulting proteins, containing 434, 433, 432, 431, 430, 429 and 427 residues per monomer, showed intermediate free energies of dissociation. Finally, three other mutations were introduced into our reference protein to establish how they influenced the equilibrium. The main importance of this work is it shows that for homo-multimeric proteins a small change in the standard Gibbs interaction energy between subunits can have major physiological effects.

  17. Crystal Structure of Acivicin-Inhibited [gamma]-Glutamyltranspeptidase Reveals Critical Roles for Its C-Terminus in Autoprocessing and Catalysis

    Energy Technology Data Exchange (ETDEWEB)

    Williams, Kristin; Cullati, Sierra; Sand, Aaron; Biterova, Ekaterina I.; Barycki, Joseph J.; (UNL)

    2009-03-27

    Helicobacter pylori {gamma}-glutamyltranspeptidase (HpGT) is a general {gamma}-glutamyl hydrolase and a demonstrated virulence factor. The enzyme confers a growth advantage to the bacterium, providing essential amino acid precursors by initiating the degradation of extracellular glutathione and glutamine. HpGT is a member of the N-terminal nucleophile (Ntn) hydrolase superfamily and undergoes autoprocessing to generate the active form of the enzyme. Acivicin is a widely used {gamma}-glutamyltranspeptidase inhibitor that covalently modifies the enzyme, but its precise mechanism of action remains unclear. The time-dependent inactivation of HpGT exhibits a hyperbolic dependence on acivicin concentration with k{sub max} = 0.033 {+-} 0.006 s{sup -1} and K{sub I} = 19.7 {+-} 7.2 {micro}M. Structure determination of acivicin-modified HpGT (1.7 {angstrom}; R{sub factor} = 17.9%; R{sub free} = 20.8%) demonstrates that acivicin is accommodated within the {gamma}-glutamyl binding pocket of the enzyme. The hydroxyl group of Thr 380, the catalytic nucleophile in the autoprocessing and enzymatic reactions, displaces chloride from the acivicin ring to form the covalently linked complex. Within the acivicin-modified HpGT structure, the C-terminus of the protein becomes ordered with Phe 567 positioned over the active site. Substitution or deletion of Phe 567 leads to a >10-fold reduction in enzymatic activity, underscoring its importance in catalysis. The mobile C-terminus is positioned by several electrostatic interactions within the C-terminal region, most notably a salt bridge between Arg 475 and Glu 566. Mutational analysis reveals that Arg 475 is critical for the proper placement of the C-terminal region, the Tyr 433 containing loop, and the proposed oxyanion hole.

  18. A Listeria monocytogenes RNA helicase essential for growth and ribosomal maturation at low temperatures uses its C terminus for appropriate interaction with the ribosome.

    Science.gov (United States)

    Netterling, Sakura; Vaitkevicius, Karolis; Nord, Stefan; Johansson, Jörgen

    2012-08-01

    Listeria monocytogenes, a Gram-positive food-borne human pathogen, is able to grow at temperatures close to 0°C and is thus of great concern for the food industry. In this work, we investigated the physiological role of one DExD-box RNA helicase in Listeria monocytogenes. The RNA helicase Lmo1722 was required for optimal growth at low temperatures, whereas it was dispensable at 37°C. A Δlmo1722 strain was less motile due to downregulation of the major subunit of the flagellum, FlaA, caused by decreased flaA expression. By ribosomal fractionation experiments, it was observed that Lmo1722 was mainly associated with the 50S subunit of the ribosome. Absence of Lmo1722 decreased the fraction of 50S ribosomal subunits and mature 70S ribosomes and affected the processing of the 23S precursor rRNA. The ribosomal profile could be restored to wild-type levels in a Δlmo1722 strain expressing Lmo1722. Interestingly, the C-terminal part of Lmo1722 was redundant for low-temperature growth, motility, 23S rRNA processing, and appropriate ribosomal maturation. However, Lmo1722 lacking the C terminus showed a reduced affinity for the 50S and 70S fractions, suggesting that the C terminus is important for proper guidance of Lmo1722 to the 50S subunit. Taken together, our results show that the Listeria RNA helicase Lmo1722 is essential for growth at low temperatures, motility, and rRNA processing and is important for ribosomal maturation, being associated mainly with the 50S subunit of the ribosome.

  19. Mass balance, meteorology, area altitude distribution, glacier-surface altitude, ice motion, terminus position, and runoff at Gulkana Glacier, Alaska, 1996 balance year

    Science.gov (United States)

    March, Rod S.

    2003-01-01

    The 1996 measured winter snow, maximum winter snow, net, and annual balances in the Gulkana Glacier Basin were evaluated on the basis of meteorological, hydrological, and glaciological data. Averaged over the glacier, the measured winter snow balance was 0.87 meter on April 18, 1996, 1.1 standard deviation below the long-term average; the maximum winter snow balance, 1.06 meters, was reached on May 28, 1996; and the net balance (from August 30, 1995, to August 24, 1996) was -0.53 meter, 0.53 standard deviation below the long-term average. The annual balance (October 1, 1995, to September 30, 1996) was -0.37 meter. Area-averaged balances were reported using both the 1967 and 1993 area altitude distributions (the numbers previously given in this abstract use the 1993 area altitude distribution). Net balance was about 25 percent less negative using the 1993 area altitude distribution than the 1967 distribution. Annual average air temperature was 0.9 degree Celsius warmer than that recorded with the analog sensor used since 1966. Total precipitation catch for the year was 0.78 meter, 0.8 standard deviations below normal. The annual average wind speed was 3.5 meters per second in the first year of measuring wind speed. Annual runoff averaged 1.50 meters over the basin, 1.0 standard deviation below the long-term average. Glacier-surface altitude and ice-motion changes measured at three index sites document seasonal ice-speed and glacier-thickness changes. Both showed a continuation of a slowing and thinning trend present in the 1990s. The glacier terminus and lower ablation area were defined for 1996 with a handheld Global Positioning System survey of 126 locations spread out over about 4 kilometers on the lower glacier margin. From 1949 to 1996, the terminus retreated about 1,650 meters for an average retreat rate of 35 meters per year.

  20. Ipomoelin, a Jacalin-Related Lectin with a Compact Tetrameric Association and Versatile Carbohydrate Binding Properties Regulated by Its N Terminus

    Science.gov (United States)

    Chang, Wei-Chieh; Liu, Kai-Lun; Hsu, Fang-Ciao; Jeng, Shih-Tong; Cheng, Yi-Sheng

    2012-01-01

    Many proteins are induced in the plant defense response to biotic stress or mechanical wounding. One group is lectins. Ipomoelin (IPO) is one of the wound-inducible proteins of sweet potato (Ipomoea batatas cv. Tainung 57) and is a Jacalin-related lectin (JRL). In this study, we resolved the crystal structures of IPO in its apo form and in complex with carbohydrates such as methyl α-D-mannopyranoside (Me-Man), methyl α-D-glucopyranoside (Me-Glc), and methyl α-D-galactopyranoside (Me-Gal) in different space groups. The packing diagrams indicated that IPO might represent a compact tetrameric association in the JRL family. The protomer of IPO showed a canonical β-prism fold with 12 strands of β-sheets but with 2 additional short β-strands at the N terminus. A truncated IPO (ΔN10IPO) by removing the 2 short β-strands of the N terminus was used to reveal its role in a tetrameric association. Gel filtration chromatography confirmed IPO as a tetrameric form in solution. Isothermal titration calorimetry determined the binding constants (KA) of IPO and ΔN10IPO against various carbohydrates. IPO could bind to Me-Man, Me-Glc, and Me-Gal with similar binding constants. In contrast, ΔN10IPO showed high binding ability to Me-Man and Me-Glc but could not bind to Me-Gal. Our structural and functional analysis of IPO revealed that its compact tetrameric association and carbohydrate binding polyspecificity could be regulated by the 2 additional N-terminal β-strands. The versatile carbohydrate binding properties of IPO might play a role in plant defense. PMID:22808208

  1. Gelsolin functions as a metastasis suppressor in B16-BL6 mouse melanoma cells and requirement of the carboxyl-terminus for its effect.

    Science.gov (United States)

    Fujita, H; Okada, F; Hamada , J; Hosokawa, M; Moriuchi, T; Koya, R C; Kuzumaki, N

    2001-09-01

    Gelsolin, an actin-binding protein, is implicated as a critical regulator in cell motility. In addition, we have reported that cellular levels of gelsolin are decreased in various tumor cells, and overexpression of gelsolin by gene transfer suppresses tumorigenicity. We sought to assess the effects of gelsolin overexpression on metastasis and to determine the importance of a carboxyl-terminus that confers Ca(2+) dependency on gelsolin for effects of its overexpression. Expression vectors with cDNA encoding either full-length wild-type or His321 mutant form, isolated from a flat revertant of Ras-transformed cells and a carboxyl-terminal truncate, C-del of gelsolin, were transfected into a highly metastatic murine melanoma cell line, B16-BL6. Expression of introduced cDNA in transfectants was confirmed using Western blotting, 2-dimensional gel electrophoresis and reverse transcription-polymerase chain reaction (RT-PCR). We characterized phenotypes of transfectants, such as growth rate, colony formation in soft agar, cell motility and metastasis formation in vivo. Transfectants expressing the wild-type, His321 mutant and C-del gelsolin exhibited reduced growth ability in soft agar. Although expression of integrin beta1 or alpha4 on the cell surface of transfectants was not changed, wild-type and His321 mutant gelsolin, except for C-del gelsolin, exhibited retardation of cell spreading, reduced chemotatic migration to fibronectin and suppressed lung colonization in spontaneous metastasis assay. Gelsolin may function as a metastasis suppressor as well as a tumor suppressor gene. The carboxyl-terminus of gelsolin is important for retardation of cell spreading, reduced chemotasis and metastasis suppression. Copyright 2001 Wiley-Liss, Inc.

  2. Multisite tyrosine phosphorylation of the N-terminus of Mint1/X11α by Src kinase regulates the trafficking of amyloid precursor protein.

    Science.gov (United States)

    Dunning, Christopher J R; Black, Hannah L; Andrews, Katie L; Davenport, Elizabeth C; Conboy, Michael; Chawla, Sangeeta; Dowle, Adam A; Ashford, David; Thomas, Jerry R; Evans, Gareth J O

    2016-05-01

    Mint/X11 is one of the four neuronal trafficking adaptors that interact with amyloid precursor protein (APP) and are linked with its cleavage to generate β-amyloid peptide, a key player in the pathology of Alzheimer's disease. How APP switches between adaptors at different stages of the secretory pathway is poorly understood. Here, we show that tyrosine phosphorylation of Mint1 regulates the destination of APP. A canonical SH2-binding motif ((202) YEEI) was identified in the N-terminus of Mint1 that is phosphorylated on tyrosine by C-Src and recruits the active kinase for sequential phosphorylation of further tyrosines (Y191 and Y187). A single Y202F mutation in the Mint1 N-terminus inhibits C-Src binding and tyrosine phosphorylation. Previous studies observed that co-expression of wild-type Mint1 and APP causes accumulation of APP in the trans-Golgi. Unphosphorylatable Mint1 (Y202F) or pharmacological inhibition of Src reduced the accumulation of APP in the trans-Golgi of heterologous cells. A similar result was observed in cultured rat hippocampal neurons where Mint1(Y202F) permitted the trafficking of APP to more distal neurites than the wild-type protein. These data underline the importance of the tyrosine phosphorylation of Mint1 as a critical switch for determining the destination of APP. The regulation of amyloid precursor protein (APP) trafficking is poorly understood. We have discovered that the APP adapter, Mint1, is phosphorylated by C-Src kinase. Mint1 causes APP accumulation in the trans-Golgi network, whereas inhibition of Src or mutation of Mint1-Y202 permits APP recycling. The phosphorylation status of Mint1 could impact on the pathological trafficking of APP in Alzheimer's disease. © 2016 The Authors. Journal of Neurochemistry published by John Wiley & Sons Ltd on behalf of International Society for Neurochemistry.

  3. The dengue vector Aedes aegypti contains a functional high mobility group box 1 (HMGB1 protein with a unique regulatory C-terminus.

    Directory of Open Access Journals (Sweden)

    Fabio Schneider Ribeiro

    Full Text Available The mosquito Aedes aegypti can spread the dengue, chikungunya and yellow fever viruses. Thus, the search for key molecules involved in the mosquito survival represents today a promising vector control strategy. High Mobility Group Box (HMGB proteins are essential nuclear factors that maintain the high-order structure of chromatin, keeping eukaryotic cells viable. Outside the nucleus, secreted HMGB proteins could alert the innate immune system to foreign antigens and trigger the initiation of host defenses. In this work, we cloned and functionally characterized the HMGB1 protein from Aedes aegypti (AaHMGB1. The AaHMGB1 protein typically consists of two HMG-box DNA binding domains and an acidic C-terminus. Interestingly, AaHMGB1 contains a unique alanine/glutamine-rich (AQ-rich C-terminal region that seems to be exclusive of dipteran HMGB proteins. AaHMGB1 is localized to the cell nucleus, mainly associated with heterochromatin. Circular dichroism analyses of AaHMGB1 or the C-terminal truncated proteins revealed α-helical structures. We showed that AaHMGB1 can effectively bind and change the topology of DNA, and that the AQ-rich and the C-terminal acidic regions can modulate its ability to promote DNA supercoiling, as well as its preference to bind supercoiled DNA. AaHMGB1 is phosphorylated by PKA and PKC, but not by CK2. Importantly, phosphorylation of AaHMGB1 by PKA or PKC completely abolishes its DNA bending activity. Thus, our study shows that a functional HMGB1 protein occurs in Aedes aegypt and we provide the first description of a HMGB1 protein containing an AQ-rich regulatory C-terminus.

  4. Deletion of the distal COOH-terminus of the A2B adenosine receptor switches internalization to an arrestin- and clathrin-independent pathway and inhibits recycling.

    Science.gov (United States)

    Mundell, S J; Matharu, A-L; Nisar, S; Palmer, T M; Benovic, J L; Kelly, E

    2010-02-01

    We have investigated the effect of deletions of a postsynaptic density, disc large and zo-1 protein (PDZ) motif at the end of the COOH-terminus of the rat A(2B) adenosine receptor on intracellular trafficking following long-term exposure to the agonist 5'-(N-ethylcarboxamido)-adenosine. The trafficking of the wild type A(2B) adenosine receptor and deletion mutants expressed in Chinese hamster ovary cells was studied using an enzyme-linked immunosorbent assay in combination with immunofluorescence microscopy. The wild type A(2B) adenosine receptor and deletion mutants were all extensively internalized following prolonged treatment with NECA. The intracellular compartment through which the Gln(325)-stop receptor mutant, which lacks the Type II PDZ motif found in the wild type receptor initially trafficked was not the same as the wild type receptor. Expression of dominant negative mutants of arrestin-2, dynamin or Eps-15 inhibited internalization of wild type and Leu(330)-stop receptors, whereas only dominant negative mutant dynamin inhibited agonist-induced internalization of Gln(325)-stop, Ser(326)-stop and Phe(328)-stop receptors. Following internalization, the wild type A(2B) adenosine receptor recycled rapidly to the cell surface, whereas the Gln(325)-stop receptor did not recycle. Deletion of the COOH-terminus of the A(2B) adenosine receptor beyond Leu(330) switches internalization from an arrestin- and clathrin-dependent pathway to one that is dynamin dependent but arrestin and clathrin independent. The presence of a Type II PDZ motif appears to be essential for arrestin- and clathrin-dependent internalization, as well as recycling of the A(2B) adenosine receptor following prolonged agonist addition.

  5. Involvement of 5-HT(2) serotonergic receptors of the nucleus raphe magnus and nucleus reticularis gigantocellularis/paragigantocellularis complex neural networks in the antinociceptive phenomenon that follows the post-ictal immobility syndrome.

    Science.gov (United States)

    de Oliveira, Rithiele Cristina; de Oliveira, Ricardo; Ferreira, Célio Marcos Dos Reis; Coimbra, Norberto Cysne

    2006-09-01

    The post-ictal immobility syndrome is followed by a significant increase in the nociceptive thresholds in animals and men. In this interesting post-ictal behavioral response, endogenous opioid peptides-mediated mechanisms, as well as cholinergic-mediated antinociceptive processes, have been suggested. However, considering that many serotonergic descending pathways have been implicated in antinociceptive reactions, the aim of the present work is to investigate the involvement of 5-HT(2)-serotonergic receptor subfamily in the post-ictal antinociception. The analgesia was measured by the tail-flick test in seven or eight Wistar rats per group. Convulsions were followed by statistically significant increase in the tail-flick latencies (TFL), at least for 120 min of the post-ictal period. Male Wistar rats were submitted to stereotaxic surgery for introduction of a guide-cannula in the rhombencephalon, aiming either the nucleus raphe magnus (NRM) or the gigantocellularis complex. In independent groups of animals, these nuclei were neurochemically lesioned with a unilateral microinjection of ibotenic acid (1.0 microg/0.2 microL). The neuronal damage of either the NRM or nucleus reticularis gigantocellularis/paragigantocellularis complex decreased the post-ictal analgesia. Also, in other independent groups, central administration of ritanserin (5.0 microg/0.2 microL) or physiological saline into each of the reticular formation nuclei studied caused a statistically significant decrease in the TFL of seizing animals, as compared to controls, in all post-ictal periods studied. These results indicate that serotonin input-connected neurons of the pontine and medullarly reticular nuclei may be involved in the post-ictal analgesia.

  6. Anxiogenic drug administration and elevated plus-maze exposure in rats activate populations of relaxin-3 neurons in the nucleus incertus and serotonergic neurons in the dorsal raphe nucleus.

    Science.gov (United States)

    Lawther, A J; Clissold, M L; Ma, S; Kent, S; Lowry, C A; Gundlach, A L; Hale, M W

    2015-09-10

    Anxiety is a complex and adaptive emotional state controlled by a distributed and interconnected network of brain regions, and disruption of these networks is thought to give rise to the behavioral symptoms associated with anxiety disorders in humans. The dorsal raphe nucleus (DR), which contains the majority of forebrain-projecting serotonergic neurons, is implicated in the control of anxiety states and anxiety-related behavior via neuromodulatory effects on these networks. Relaxin-3 is the native neuropeptide ligand for the Gi/o-protein-coupled receptor, RXFP3, and is primarily expressed in the nucleus incertus (NI), a tegmental region immediately caudal to the DR. RXFP3 activation has been shown to modulate anxiety-related behavior in rodents, and RXFP3 mRNA is expressed in the DR. In this study, we examined the response of relaxin-3-containing neurons in the NI and serotonergic neurons in the DR following pharmacologically induced anxiety and exposure to an aversive environment. We administered the anxiogenic drug FG-7142 or vehicle to adult male Wistar rats and, 30 min later, exposed them to either the elevated plus-maze or home cage control conditions. Immunohistochemical detection of c-Fos was used to determine activation of serotonergic neurons in the DR and relaxin-3 neurons in the NI, measured 2h following drug injection. Analysis revealed that FG-7142 administration and exposure to the elevated plus-maze are both associated with an increase in c-Fos expression in relaxin-3-containing neurons in the NI and in serotonergic neurons in dorsal and ventrolateral regions of the DR. These data are consistent with the hypothesis that relaxin-3 systems in the NI and serotonin systems in the DR interact to form part of a network involved in the control of anxiety-related behavior. Copyright © 2015 IBRO. Published by Elsevier Ltd. All rights reserved.

  7. Creating and Viewing HTML Files with Python

    Directory of Open Access Journals (Sweden)

    William J. Turkel

    2012-07-01

    Full Text Available This lesson uses Python to create and view an HTML file. If you write programs that output HTML, you can use any browser to look at your results. This is especially convenient if your program is automatically creating hyperlinks or graphic entities like charts and diagrams. Here you will learn how to create HTML files with Python scripts, and how to use Python to automatically open an HTML file in Firefox.

  8. Creating and Viewing HTML Files with Python

    OpenAIRE

    William J. Turkel; Adam Crymble

    2012-01-01

    This lesson uses Python to create and view an HTML file. If you write programs that output HTML, you can use any browser to look at your results. This is especially convenient if your program is automatically creating hyperlinks or graphic entities like charts and diagrams. Here you will learn how to create HTML files with Python scripts, and how to use Python to automatically open an HTML file in Firefox.

  9. The C Terminus of the Core β-Ladder Domain in Japanese Encephalitis Virus Nonstructural Protein 1 Is Flexible for Accommodation of Heterologous Epitope Fusion.

    Science.gov (United States)

    Yen, Li-Chen; Liao, Jia-Teh; Lee, Hwei-Jen; Chou, Wei-Yuan; Chen, Chun-Wei; Lin, Yi-Ling; Liao, Ching-Len

    2016-02-01

    NS1 is the only nonstructural protein that enters the lumen of the endoplasmic reticulum (ER), where NS1 is glycosylated, forms a dimer, and is subsequently secreted during flavivirus replication as dimers or hexamers, which appear to be highly immunogenic to the infected host, as protective immunity can be elicited against homologous flavivirus infections. Here, by using a trans-complementation assay, we identified the C-terminal end of NS1 derived from Japanese encephalitis virus (JEV), which was more flexible than other regions in terms of housing foreign epitopes without a significant impact on virus replication. This mapped flexible region is located in the conserved tip of the core β-ladder domain of the multimeric NS1 structure and is also known to contain certain linear epitopes, readily triggering specific antibody responses from the host. Despite becoming attenuated, recombinant JEV with insertion of a neutralizing epitope derived from enterovirus 71 (EV71) into the C-terminal end of NS1 not only could be normally released from infected cells, but also induced dual protective immunity for the host to counteract lethal challenge with either JEV or EV71 in neonatal mice. These results indicated that the secreted multimeric NS1 of flaviviruses may serve as a natural protein carrier to render epitopes of interest more immunogenic in the C terminus of the core β-ladder domain. The positive-sense RNA genomes of mosquito-borne flaviviruses appear to be flexible in terms of accommodating extra insertions of short heterologous antigens into their virus genes. Here, we illustrate that the newly identified C terminus of the core β-ladder domain in NS1 could be readily inserted into entities such as EV71 epitopes, and the resulting NS1-epitope fusion proteins appeared to maintain normal virus replication, secretion ability, and multimeric formation from infected cells. Nonetheless, such an insertion attenuated the recombinant JEV in mice, despite having retained

  10. Crystal structure of the karyopherin Kap121p bound to the extreme C-terminus of the protein phosphatase Cdc14p

    Energy Technology Data Exchange (ETDEWEB)

    Kobayashi, Junya [Division of Biological Science, Graduate School of Science, Nagoya University (Japan); Hirano, Hidemi [Division of Biological Science, Graduate School of Science, Nagoya University (Japan); Structural Biology Research Center, Graduate School of Science, Nagoya University (Japan); Matsuura, Yoshiyuki, E-mail: matsuura.yoshiyuki@d.mbox.nagoya-u.ac.jp [Division of Biological Science, Graduate School of Science, Nagoya University (Japan); Structural Biology Research Center, Graduate School of Science, Nagoya University (Japan)

    2015-07-31

    In Saccharomyces cerevisiae, the protein phosphatase Cdc14p is an antagonist of mitotic cyclin-dependent kinases and is a key regulator of late mitotic events such as chromosome segregation, spindle disassembly and cytokinesis. The activity of Cdc14p is controlled by cell-cycle dependent changes in its association with its competitive inhibitor Net1p (also known as Cfi1p) in the nucleolus. For most of the cell cycle up to metaphase, Cdc14p is sequestered in the nucleolus in an inactive state. During anaphase, Cdc14p is released from Net1p, spreads into the nucleus and cytoplasm, and dephosphorylates key mitotic targets. Although regulated nucleocytoplasmic shuttling of Cdc14p has been suggested to be important for exit from mitosis, the mechanism underlying Cdc14p nuclear trafficking remains poorly understood. Here we show that the C-terminal region (residues 517–551) of Cdc14p can function as a nuclear localization signal (NLS) in vivo and also binds to Kap121p (also known as Pse1p), an essential nuclear import carrier in yeast, in a Gsp1p-GTP-dependent manner in vitro. Moreover we report a crystal structure, at 2.4 Å resolution, of Kap121p bound to the C-terminal region of Cdc14p. The structure and structure-based mutational analyses suggest that either the last five residues at the extreme C-terminus of Cdc14p (residues 547–551; Gly-Ser-Ile-Lys-Lys) or adjacent residues with similar sequence (residues 540–544; Gly-Gly-Ile-Arg-Lys) can bind to the NLS-binding site of Kap121p, with two residues (Ile in the middle and Lys at the end of the five residues) of Cdc14p making key contributions to the binding specificity. Based on comparison with other structures of Kap121p-ligand complexes, we propose “IK-NLS” as an appropriate term to refer to the Kap121p-specific NLS. - Highlights: • The C-terminus of Cdc14p binds to Kap121p in a Gsp1p-GTP-dependent manner. • The crystal structure of Kap121p-Cdc14p complex is determined. • The structure reveals how

  11. Crystal structure of the karyopherin Kap121p bound to the extreme C-terminus of the protein phosphatase Cdc14p

    International Nuclear Information System (INIS)

    Kobayashi, Junya; Hirano, Hidemi; Matsuura, Yoshiyuki

    2015-01-01

    In Saccharomyces cerevisiae, the protein phosphatase Cdc14p is an antagonist of mitotic cyclin-dependent kinases and is a key regulator of late mitotic events such as chromosome segregation, spindle disassembly and cytokinesis. The activity of Cdc14p is controlled by cell-cycle dependent changes in its association with its competitive inhibitor Net1p (also known as Cfi1p) in the nucleolus. For most of the cell cycle up to metaphase, Cdc14p is sequestered in the nucleolus in an inactive state. During anaphase, Cdc14p is released from Net1p, spreads into the nucleus and cytoplasm, and dephosphorylates key mitotic targets. Although regulated nucleocytoplasmic shuttling of Cdc14p has been suggested to be important for exit from mitosis, the mechanism underlying Cdc14p nuclear trafficking remains poorly understood. Here we show that the C-terminal region (residues 517–551) of Cdc14p can function as a nuclear localization signal (NLS) in vivo and also binds to Kap121p (also known as Pse1p), an essential nuclear import carrier in yeast, in a Gsp1p-GTP-dependent manner in vitro. Moreover we report a crystal structure, at 2.4 Å resolution, of Kap121p bound to the C-terminal region of Cdc14p. The structure and structure-based mutational analyses suggest that either the last five residues at the extreme C-terminus of Cdc14p (residues 547–551; Gly-Ser-Ile-Lys-Lys) or adjacent residues with similar sequence (residues 540–544; Gly-Gly-Ile-Arg-Lys) can bind to the NLS-binding site of Kap121p, with two residues (Ile in the middle and Lys at the end of the five residues) of Cdc14p making key contributions to the binding specificity. Based on comparison with other structures of Kap121p-ligand complexes, we propose “IK-NLS” as an appropriate term to refer to the Kap121p-specific NLS. - Highlights: • The C-terminus of Cdc14p binds to Kap121p in a Gsp1p-GTP-dependent manner. • The crystal structure of Kap121p-Cdc14p complex is determined. • The structure reveals how

  12. Application of Nanotrap technology for high sensitivity measurement of urinary outer surface protein A carboxyl-terminus domain in early stage Lyme borreliosis.

    Science.gov (United States)

    Magni, Ruben; Espina, Benjamin H; Shah, Ketul; Lepene, Benjamin; Mayuga, Christine; Douglas, Temple A; Espina, Virginia; Rucker, Sally; Dunlap, Ross; Petricoin, Emanuel F Iii; Kilavos, Mary Frekko; Poretz, Donald M; Irwin, Gilbert R; Shor, Samuel M; Liotta, Lance A; Luchini, Alessandra

    2015-11-04

    Prompt antibiotic treatment of early stage Lyme borreliosis (LB) prevents progression to severe multisystem disease. There is a clinical need to improve the diagnostic specificity of early stage Lyme assays in the period prior to the mounting of a robust serology response. Using a novel analyte harvesting nanotechnology, Nanotrap particles, we evaluated urinary Borrelia Outer surface protein A (OspA) C-terminus peptide in early stage LB before and after treatment, and in patients suspected of late stage disseminated LB. We employed Nanotrap particles to concentrate urinary OspA and used a highly specific anti-OspA monoclonal antibody (mAb) as a detector of the C-terminus peptides. We mapped the mAb epitope to a narrow specific OspA C-terminal domain OspA236-239 conserved across infectious Borrelia species but with no homology to human proteins and no cross-reactivity with relevant viral and non-Borrelia bacterial proteins. 268 urine samples from patients being evaluated for all categories of LB were collected in a LB endemic area. The urinary OspA assay, blinded to outcome, utilized Nanotrap particle pre-processing, western blotting to evaluate the OspA molecular size, and OspA peptide competition for confirmation. OspA test characteristics: sensitivity 1.7 pg/mL (lowest limit of detection), % coefficient of variation (CV) = 8 %, dynamic range 1.7-30 pg/mL. Pre-treatment, 24/24 newly diagnosed patients with an erythema migrans (EM) rash were positive for urinary OspA while false positives for asymptomatic patients were 0/117 (Chi squared p antibiotic therapy, 10/10 were positive for urinary OspA. Urinary OspA of 8/8 patients switched from detectable to undetectable following symptom resolution post-treatment. Specificity of the urinary OspA test for the clinical symptoms was 40/40. Specificity of the urinary OspA antigen test for later serology outcome was 87.5 % (21 urinary OspA positive/24 serology positive, Chi squared p = 4.072e(-15)). 41 of 100 patients under

  13. Creating Gaze Annotations in Head Mounted Displays

    DEFF Research Database (Denmark)

    Mardanbeigi, Diako; Qvarfordt, Pernilla

    2015-01-01

    To facilitate distributed communication in mobile settings, we developed GazeNote for creating and sharing gaze annotations in head mounted displays (HMDs). With gaze annotations it possible to point out objects of interest within an image and add a verbal description. To create an annota- tion...

  14. Creating Safe Spaces for Music Learning

    Science.gov (United States)

    Hendricks, Karin S.; Smith, Tawnya D.; Stanuch, Jennifer

    2014-01-01

    This article offers a practical model for fostering emotionally safe learning environments that instill in music students a positive sense of self-belief, freedom, and purpose. The authors examine the implications for music educators of creating effective learning environments and present recommendations for creating a safe space for learning,…

  15. The N-terminus of RPA large subunit and its spatial position are important for the 5'->3' resection of DNA double-strand breaks.

    Science.gov (United States)

    Tammaro, Margaret; Liao, Shuren; McCane, Jill; Yan, Hong

    2015-10-15

    The first step of homology-dependent repair of DNA double-strand breaks (DSBs) is the resection of the 5' strand to generate 3' ss-DNA. Of the two major nucleases responsible for resection, EXO1 has intrinsic 5'->3' directionality, but DNA2 does not. DNA2 acts with RecQ helicases such as the Werner syndrome protein (WRN) and the heterotrimeric eukaryotic ss-DNA binding protein RPA. We have found that the N-terminus of the RPA large subunit (RPA1N) interacts with both WRN and DNA2 and is essential for stimulating WRN's 3'->5' helicase activity and DNA2's 5'->3' ss-DNA exonuclease activity. A mutant RPA complex that lacks RPA1N is unable to support resection in Xenopus egg extracts and human cells. Furthermore, relocating RPA1N to the middle subunit but not to the small subunit causes severe defects in stimulating DNA2 and WRN and in supporting resection. Together, these findings suggest that RPA1N and its spatial position are critical for restricting the directionality of the WRN-DNA2 resection pathway. © The Author(s) 2015. Published by Oxford University Press on behalf of Nucleic Acids Research.

  16. The N-terminus of RPA large subunit and its spatial position are important for the 5′->3′ resection of DNA double-strand breaks

    Science.gov (United States)

    Tammaro, Margaret; Liao, Shuren; McCane, Jill; Yan, Hong

    2015-01-01

    The first step of homology-dependent repair of DNA double-strand breaks (DSBs) is the resection of the 5′ strand to generate 3′ ss-DNA. Of the two major nucleases responsible for resection, EXO1 has intrinsic 5′->3′ directionality, but DNA2 does not. DNA2 acts with RecQ helicases such as the Werner syndrome protein (WRN) and the heterotrimeric eukaryotic ss-DNA binding protein RPA. We have found that the N-terminus of the RPA large subunit (RPA1N) interacts with both WRN and DNA2 and is essential for stimulating WRN's 3′->5′ helicase activity and DNA2's 5′->3′ ss-DNA exonuclease activity. A mutant RPA complex that lacks RPA1N is unable to support resection in Xenopus egg extracts and human cells. Furthermore, relocating RPA1N to the middle subunit but not to the small subunit causes severe defects in stimulating DNA2 and WRN and in supporting resection. Together, these findings suggest that RPA1N and its spatial position are critical for restricting the directionality of the WRN-DNA2 resection pathway. PMID:26227969

  17. The carboxyl-terminus directs TAF(I)48 to the nucleus and nucleolus and associates with multiple nuclear import receptors.

    Science.gov (United States)

    Dynes, Joseph L; Xu, Shuping; Bothner, Sarah; Lahti, Jill M; Hori, Roderick T

    2004-03-01

    The protein complex Selectivity Factor 1, composed of TBP, TAF(I)48, TAF(I)63 and TAF(I)110, is required for rRNA transcription by RNA polymerase I in the nucleolus. The steps involved in targeting Selectivity Factor 1 will be dependent on the transport pathways that are used and the localization signals that direct this trafficking. In order to investigate these issues, we characterized human TAF(I)48, a subunit of Selectivity Factor 1. By domain analysis of TAF(I)48, the carboxyl-terminal 51 residues were found to be required for the localization of TAF(I)48, as well as sufficient to direct Green Fluorescent Protein to the nucleus and nucleolus. The carboxyl-terminus of TAF(I)48 also has the ability to associate with multiple members of the beta-karyopherin family of nuclear import receptors, including importin beta (karyopherin beta1), transportin (karyopherin beta2) and RanBP5 (karyopherin beta3), in a Ran-dependent manner. This property of interacting with multiple beta-karyopherins has been previously reported for the nuclear localization signals of some ribosomal proteins that are likewise directed to the nucleolus. This study identifies the first nuclear import sequence identified within the TBP-Associated Factor subunits of Selectivity Factor 1.

  18. Is the Okavango Delta the terminus of the East African Rift System? Towards a new geodynamic model: Geodetic study and geophysical review

    Science.gov (United States)

    Pastier, Anne-Morwenn; Dauteuil, Olivier; Murray-Hudson, Michael; Moreau, Frédérique; Walpersdorf, Andrea; Makati, Kaelo

    2017-08-01

    The Okavango Graben (OG) has been considered as the terminus of the southwestern branch of the East African Rift System (EARS) since the 1970s based on fault morphology and early seismic and geophysical data. Thus it has been assumed to be an incipient rifting zone, analogous to the early stage of mature rifts in the EARS. Recent geodetic data and geophysical studies in the area bring new insights into the local crust and lithosphere, mantle activity and fault activity. In this study, we computed the velocities for three permanent GPS stations surrounding the graben and undertook a review of the new geophysical data available for the area. The northern and southern blocks of the graben show an exclusively low strike-slip displacement rate of about 1mm/year, revealing the transtensional nature of this basin. The seismic record of central and southern Africa was found to be instrumentally biased for the events recorded before 2004 and the OG may not represent the most seismically active area in Botswana anymore. Moreover, no significant lithosphere and crustal thinning is found in the tectonic structure nor any strong negative Bouguer anomaly and surface heat flux. Thus the OG does not match the classical model for a rifting zone. We propose a new geodynamic model for the deformation observed west of the EARS based on accommodation of far-field deformation due to the differential extension rates of the EARS and the displacement of the Kalahari craton relative to the Nubian plate.

  19. A novel transcript of cyclin-dependent kinase-like 5 (CDKL5) has an alternative C-terminus and is the predominant transcript in brain.

    Science.gov (United States)

    Williamson, Sarah L; Giudici, Laura; Kilstrup-Nielsen, Charlotte; Gold, Wendy; Pelka, Gregory J; Tam, Patrick P L; Grimm, Andrew; Prodi, Dionigio; Landsberger, Nicoletta; Christodoulou, John

    2012-02-01

    The X-linked cyclin-dependent kinase-like 5 (CDKL5) gene is an important molecular determinant of early-onset intractable seizures with infantile spasms and Rett syndrome-like phenotype. The gene encodes a kinase that may influence components of molecular pathways associated with MeCP2. In humans there are two previously reported splice variants that differ in the 5' untranslated exons and produce the same 115 kDa protein. Furthermore, very recently, a novel transcript including a novel exon (16b) has been described. By aligning both the human and mouse CDKL5 proteins to the orthologs of other species, we identified a theoretical 107 kDa isoform with an alternative C-terminus that terminates in intron 18. In human brain and all other tissues investigated except the testis, this novel isoform is the major CDKL5 transcript. The detailed characterisation of this novel isoform of CDKL5 reveals functional and subcellular localisation attributes that overlap greatly, but not completely, with that of the previously studied human CDKL5 protein. Considering its predominant expression in the human and mouse brain, we believe that this novel isoform is likely to be of primary pathogenic importance in human diseases associated with CDKL5 deficiency, and suggest that screening of the related intronic sequence should be included in the molecular genetic analyses of patients with a suggestive clinical phenotype.

  20. Identification of a rare point mutation at C-terminus of merozoite surface antigen-1 gene of Plasmodium falciparum in eastern Indian isolates.

    Science.gov (United States)

    Raj, Dipak Kumar; Das, Bibhu Ranjan; Dash, A P; Supakar, Prakash C

    2004-01-01

    Merozoite surface antigen-1 (MSA-1) of Plasmodium falciparum is highly immunogenic in human. Several studies suggest that MSA-1 protein is an effective target for a protective immune response. Attempt has been made to find new point mutations by analyzing 244 bp [codon 1655(R) to 1735 (I)] relatively conserved C-terminus region of MSA-1 gene in 125 isolates. This region contains two EGF like domains, which are involved in generating protective immune response in human. Point mutations in this region are very much important in view of vaccine development. Searching of mutational hot spots in MSA-1 protein by sequencing method in a representative number of isolates is quite critical and expensive. Therefore, in this study slot blot and PCR-SSCP method have been used to find out new mutations in the individual isolates showing alterations in the mobility of DNA fragment. Sequencing of the altered bands from the SSCP gel shows a rare non-synonymous point mutation in 7 (5.6%) of the 125 isolates at amino acid position 1704 of MSA-1 gene where isoleucine is replaced by valine.

  1. Direct binding of the N-terminus of HTLV-1 tax oncoprotein to cyclin-dependent kinase 4 is a dominant path to stimulate the kinase activity.

    Science.gov (United States)

    Li, Junan; Li, Hongyuan; Tsai, Ming-Daw

    2003-06-10

    The involvement of Tax oncoprotein in the INK4-CDK4/6-Rb pathway has been regarded as a key factor for immortalization and transformation of human T-cell leukemia virus 1 (HTLV-1) infected cells. In both p16 -/- and +/+ cells, expression of Tax has been correlated with an increase in CDK4 activity, which subsequently increases the phosphorylation of Rb and drives the infected cells into cell cycle progression. In relation to these effects, Tax has been shown to interact with two components of the INK4-CDK4/6-Rb pathway, p16 and cyclin D(s). While Tax competes with CDK4 for p16 binding, thus suppressing p16 inhibition of CDK4, Tax also binds to cyclin D(s) with concomitant increases in both CDK4 activity and the phosphorylation of cyclin D(s). Here we show that both Tax and residues 1-40 of the N-terminus of Tax, Tax40N, bind to and activate CDK4 in vitro. In the presence of INK4 proteins, binding of Tax and Tax40N to CDK4 counteracts against the inhibition of p16 and p18 and acts as the major path to regulate Tax-mediated activation of CDK4. We also report that Tax40N retains the transactivation ability. These results of in vitro studies demonstrate a potentially novel, p16-independent route to regulate CDK4 activity by the Tax oncoprotein in HTLV-1 infected cells.

  2. Dependence creating properties of lipotropin C-fragment (β-endorphin): Evidence for its internal control of behavior

    NARCIS (Netherlands)

    Ree, J.M. van; Smyth, D.G.; Colpaert, F.C.

    1979-01-01

    The C-fragment of lipotropin (β-endorphin) possesses reinforcing properties, in that this peptide, like heroin, induced intraventricular self-administering behavior in drug naive rats. Only mild behavioral signs reminiscent of physical dependence were present. After injection into the nucleus raphé

  3. Creating mobile apps with Appcelerator Titanium

    CERN Document Server

    Brousseau, Christian

    2013-01-01

    Creating Mobile Apps with Appcelerator Titanium provides a hands-on approach and working examples on creating apps and games as well as embedding them onto a social networking website. Developers can then move on from there to develop their own applications based on the ones they have developed throughout the course of this book.""Creating Mobile Apps with Appcelerator Titanium"" is for developers who have experience with modern languages and development environments. Also, if you are familiar with the concepts of Object-oriented Programming (OOP), reusable components, AJAX closures, and so on

  4. Creating Web Sites The Missing Manual

    CERN Document Server

    MacDonald, Matthew

    2006-01-01

    Think you have to be a technical wizard to build a great web site? Think again. For anyone who wants to create an engaging web site--for either personal or business purposes--Creating Web Sites: The Missing Manual demystifies the process and provides tools, techniques, and expert guidance for developing a professional and reliable web presence. Like every Missing Manual, you can count on Creating Web Sites: The Missing Manual to be entertaining and insightful and complete with all the vital information, clear-headed advice, and detailed instructions you need to master the task at hand. Autho

  5. Creating Future Memories: A Dialogue on Process

    DEFF Research Database (Denmark)

    Rehder, Mads Middelboe

    2016-01-01

    This contribution documents the generative method we used to create a two-day public workshop on archiving digital memory, which occurred in April 2016 at the Counterplay Festival in Aarhus. Our group members, collectively known as the Futuremaking group created two distinct experiences. The “Help...... Desk of the Future” was created as a means of acting out how unhelpful many helpdesks actually are and to imagine how algorithms may be positioning our futures for us. The Museum of Random Memory functioned as a pop-up curatorial event where participants could offer up memories, experiences...

  6. Creating Space Plasma from the Ground

    Science.gov (United States)

    2016-05-12

    AFRL-AFOSR-VA-TR-2016-0179 CREATING SPACE PLASMA FROM THE GROUND Herbert C Carlson UTAH STATE UNIVERSITY Final Report 05/12/2016 DISTRIBUTION A...DATE (DD-MM-YYYY) 05/14/2016 2. REPORT TYPE Final 3. DATES COVERED (From - To) 08/14/2012-05/14/2016 4. TITLE AND SUBTITLE Creating space plasma from...Report (2016) Creating Space Plasma from the Ground Grant FA9550-11-1-0236 AFOSR Program Manager Dr. Kent Miller PI: Herbert C. Carlson Center for

  7. A summary of the CMS Create event

    CERN Multimedia

    CERN. Geneva; GASTAL, Martin

    2016-01-01

    The maiden CMS Create event took place in November 2015 and was a huge success. The output from all the participants was fantastic. As organisers we learnt a lot and hope to build on our experience for the 2016 event!

  8. Towards Creating Sustainable Ecotourism Interventions: Practical ...

    African Journals Online (AJOL)

    2017-05-01

    May 1, 2017 ... Keywords: Tourism, Development, Livelihoods, Assets, Adaptive .... widely reported in the Ghanaian media that farmers in some ..... created an outlet for revenue which, when accrued, was invested prudently in social.

  9. Taking medicine at home - create a routine

    Science.gov (United States)

    ... page: //medlineplus.gov/ency/patientinstructions/000613.htm Taking medicine at home - create a routine To use the ... teeth. Find Ways to Help You Remember Your Medicines You can: Set the alarm on your clock, ...

  10. More Sci- than Fi, Physicists Create Antimatter

    CERN Multimedia

    Overbye, Dennis

    2002-01-01

    Physicists working in Europe announced yesterday that they had passed through nature's looking glass and had created atoms made of antimatter, or antiatoms, opening up the possibility of experiments in a realm once reserved for science fiction writers (5 pages)

  11. Creating Healthy Environments For Youth Athletes

    Science.gov (United States)

    EPA has created a presentation and companion checklist to help coaches and athletic administrators better understand the environmental health risks associated with youth sports and the steps they can follow to protect young athletes.

  12. Creating Great Neighborhoods: Density in Your Community

    Science.gov (United States)

    This report highlights nine community-led efforts to create vibrant neighborhoods through density, discusses the connections between smart growth and density, and introduces design principles to ensure that density becomes a community asset.

  13. Creating an Economically Enabling and Competitive Business ...

    International Development Research Centre (IDRC) Digital Library (Canada)

    Creating an Economically Enabling and Competitive Business Environment in the ... the scope of operations of private sector enterprises in the West Bank and Gaza. ... IWRA/IDRC webinar on climate change and adaptive water management.

  14. ICT and Pragmatism: Creating sustainable Employment for ...

    African Journals Online (AJOL)

    PROF. OLIVER OSUAGWA

    effort to create self employment, this can galvanize unexpected revolution whose consequences will be very grave. The reading .... having a significant impact on the service industries. ..... secure your investment. Additional ..... Nigeria may fail.

  15. Creating Societal Benefits and Corporate Profits

    OpenAIRE

    Raisch, Sebastian; Probst, Gilbert; Gomez, Peter; Zimmermann, Alexander

    2014-01-01

    The odds of launching a new business that creates value for both the company and the public can be improved with good planning. An in-depth analysis of how four companies created for-profit initiatives that also have high societal value suggests that each followed a similar step-by-step process to achieve what the researchers call synergistic value creation. Those steps include establishing cross-business incubators and installing multi-perspective monitoring systems.

  16. ISO 55000: Creating an asset management system.

    Science.gov (United States)

    Bradley, Chris; Main, Kevin

    2015-02-01

    In the October 2014 issue of HEJ, Keith Hamer, group vice-president, Asset Management & Engineering at Sodexo, and marketing director at Asset Wisdom, Kevin Main, argued that the new ISO 55000 standards present facilities managers with an opportunity to create 'a joined-up, whole lifecycle approach' to managing and delivering value from assets. In this article, Kevin Main and Chris Bradley, who runs various asset management projects, examine the process of creating an asset management system.

  17. Creating opioid dependence in the emergency department.

    Science.gov (United States)

    Upadhye, Suneel

    2018-01-01

    Clinical question What is the risk of creating opioid dependence from an ED opioid prescription? Article chosen Barnett ML, Olenski AR, Jena AB. Opioid-prescribing patterns of emergency physicians and risk of long-term use. N Engl J Med 2017;376:663-73, doi:10.1056/NEJMsa1610524. This study examined the risk of creating long-term opioid dependence from a prescription written in an opioid-naive patient in the ED.

  18. Creating Math Videos: Comparing Platforms and Software

    Science.gov (United States)

    Abbasian, Reza O.; Sieben, John T.

    2016-01-01

    In this paper we present a short tutorial on creating mini-videos using two platforms--PCs and tablets such as iPads--and software packages that work with these devices. Specifically, we describe the step-by-step process of creating and editing videos using a Wacom Intuos pen-tablet plus Camtasia software on a PC platform and using the software…

  19. and C-terminus of

    Indian Academy of Sciences (India)

    amino acids 108–120) was expressed from Potato virus X (PVX)-based vector pGR106 as N- or C-terminal fusion with the PVX coat protein (PVX CP) in transgenic Nicotiana benthamiana plants. The fusion protein L2108-120-PVX CP was ...

  20. The C-terminus of H-Ras as a target for the covalent binding of reactive compounds modulating Ras-dependent pathways.

    Directory of Open Access Journals (Sweden)

    Clara L Oeste

    2011-01-01

    Full Text Available Ras proteins are crucial players in differentiation and oncogenesis and constitute important drug targets. The localization and activity of Ras proteins are highly dependent on posttranslational modifications at their C-termini. In addition to an isoprenylated cysteine, H-Ras, but not other Ras proteins, possesses two cysteine residues (C181 and C184 in the C-terminal hypervariable domain that act as palmitoylation sites in cells. Cyclopentenone prostaglandins (cyPG are reactive lipidic mediators that covalently bind to H-Ras and activate H-Ras dependent pathways. Dienone cyPG, such as 15-deoxy-Δ(12,14-PGJ(2 (15d-PGJ(2 and Δ(12-PGJ(2 selectively bind to the H-Ras hypervariable domain. Here we show that these cyPG bind simultaneously C181 and C184 of H-Ras, thus potentially altering the conformational tendencies of the hypervariable domain. Based on these results, we have explored the capacity of several bifunctional cysteine reactive small molecules to bind to the hypervariable domain of H-Ras proteins. Interestingly, phenylarsine oxide (PAO, a widely used tyrosine phosphatase inhibitor, and dibromobimane, a cross-linking agent used for cysteine mapping, effectively bind H-Ras hypervariable domain. The interaction of PAO with H-Ras takes place in vitro and in cells and blocks modification of H-Ras by 15d-PGJ(2. Moreover, PAO treatment selectively alters H-Ras membrane partition and the pattern of H-Ras activation in cells, from the plasma membrane to endomembranes. These results identify H-Ras as a novel target for PAO. More importantly, these observations reveal that small molecules or reactive intermediates interacting with spatially vicinal cysteines induce intramolecular cross-linking of H-Ras C-terminus potentially contributing to the modulation of Ras-dependent pathways.

  1. Deletion of the nuclear localization sequences and C-terminus of PTHrP impairs embryonic mammary development but also inhibits PTHrP production.

    Directory of Open Access Journals (Sweden)

    Kata Boras-Granic

    Full Text Available Parathyroid hormone-related protein (PTHrP can be secreted from cells and interact with its receptor, the Type 1 PTH/PTHrP Receptor (PTHR1 in an autocrine, paracrine or endocrine fashion. PTHrP can also remain inside cells and be transported into the nucleus, where its functions are unclear, although recent experiments suggest that it may broadly regulate cell survival and senescence. Disruption of either the PTHrP or PTHR1 gene results in many abnormalities including a failure of embryonic mammary gland development in mice and in humans. In order to examine the potential functions of nuclear PTHrP in the breast, we examined mammary gland development in PTHrP (1-84 knock-in mice, which express a mutant form of PTHrP that lacks the C-terminus and nuclear localization signals and which can be secreted but cannot enter the nucleus. Interestingly, we found that PTHrP (1-84 knock-in mice had defects in mammary mesenchyme differentiation and mammary duct outgrowth that were nearly identical to those previously described in PTHrP-/- and PTHR1-/- mice. However, the mammary buds in PTHrP (1-84 knock-in mice had severe reductions in mutant PTHrP mRNA levels, suggesting that the developmental defects were due to insufficient production of PTHrP by mammary epithelial cells and not loss of PTHrP nuclear function. Examination of the effects of nuclear PTHrP in the mammary gland in vivo will require the development of alternative animal models.

  2. Staphylococcus aureus SdrE captures complement factor H's C-terminus via a novel 'close, dock, lock and latch' mechanism for complement evasion.

    Science.gov (United States)

    Zhang, Yingjie; Wu, Minhao; Hang, Tianrong; Wang, Chengliang; Yang, Ye; Pan, Weimin; Zang, Jianye; Zhang, Min; Zhang, Xuan

    2017-05-04

    Complement factor H (CFH) is a soluble complement regulatory protein essential for the down-regulation of the alternative pathway on interaction with specific markers on the host cell surface. It recognizes the complement component 3b (C3b) and 3d (C3d) fragments in addition to self cell markers (i.e. glycosaminoglycans, sialic acid) to distinguish host cells that deserve protection from pathogens that should be eliminated. The Staphylococcus aureus surface protein serine-aspartate repeat protein E (SdrE) was previously reported to bind human CFH as an immune-evasion tactic. However, the molecular mechanism underlying SdrE-CFH-mediated immune evasion remains unknown. In the present study, we identified a novel region at CFH's C-terminus (CFH 1206-1226 ), which binds SdrE N2 and N3 domains (SdrE N2N3 ) with high affinity, and determined the crystal structures of apo-SdrE N2N3 and the SdrE N2N3 -CFH 1206-1226 complex. Comparison of the structure of the CFH-SdrE complex with other CFH structures reveals that CFH's C-terminal tail flips from the main body to insert into the ligand-binding groove of SdrE. In addition, SdrE N2N3 adopts a 'close' state in the absence of CFH, which undergoes a large conformational change on CFH binding, suggesting a novel 'close, dock, lock and latch' (CDLL) mechanism for SdrE to recognize its ligand. Our findings imply that SdrE functions as a 'clamp' to capture CFH's C-terminal tail via a unique CDLL mechanism and sequesters CFH on the surface of S. aureus for complement evasion. © 2017 The Author(s).

  3. Staphylococcus aureus SdrE captures complement factor H's C-terminus via a novel ‘close, dock, lock and latch' mechanism for complement evasion

    Science.gov (United States)

    Zhang, Yingjie; Wu, Minhao; Hang, Tianrong; Wang, Chengliang; Yang, Ye; Pan, Weimin; Zang, Jianye

    2017-01-01

    Complement factor H (CFH) is a soluble complement regulatory protein essential for the down-regulation of the alternative pathway on interaction with specific markers on the host cell surface. It recognizes the complement component 3b (C3b) and 3d (C3d) fragments in addition to self cell markers (i.e. glycosaminoglycans, sialic acid) to distinguish host cells that deserve protection from pathogens that should be eliminated. The Staphylococcus aureus surface protein serine–aspartate repeat protein E (SdrE) was previously reported to bind human CFH as an immune-evasion tactic. However, the molecular mechanism underlying SdrE–CFH-mediated immune evasion remains unknown. In the present study, we identified a novel region at CFH's C-terminus (CFH1206–1226), which binds SdrE N2 and N3 domains (SdrEN2N3) with high affinity, and determined the crystal structures of apo-SdrEN2N3 and the SdrEN2N3–CFH1206–1226 complex. Comparison of the structure of the CFH–SdrE complex with other CFH structures reveals that CFH's C-terminal tail flips from the main body to insert into the ligand-binding groove of SdrE. In addition, SdrEN2N3 adopts a ‘close’ state in the absence of CFH, which undergoes a large conformational change on CFH binding, suggesting a novel ‘close, dock, lock and latch' (CDLL) mechanism for SdrE to recognize its ligand. Our findings imply that SdrE functions as a ‘clamp' to capture CFH's C-terminal tail via a unique CDLL mechanism and sequesters CFH on the surface of S. aureus for complement evasion. PMID:28258151

  4. NMR structures of anti-HIV D-peptides derived from the N-terminus of viral chemokine vMIP-II

    International Nuclear Information System (INIS)

    Mori, Mayuko; Liu Dongxiang; Kumar, Santosh; Huang Ziwei

    2005-01-01

    The viral macrophage inflammatory protein-II (vMIP-II) encoded by Kaposi's sarcoma-associated herpesvirus has unique biological activities in that it blocks the cell entry by several different human immunodeficiency virus type 1 (HIV-1) strains via chemokine receptors including CXCR4 and CCR5. In this paper, we report the solution structure of all-D-amino acid peptides derived from the N-terminus of vMIP-II, which have been shown to have strong CXCR4 binding activity and potently inhibit HIV-1 entry via CXCR4, by using long mixing time two-dimensional nuclear Overhauser enhancement spectroscopy experiments. Both of all-D-peptides vMIP-II (1-10) and vMIP-II (1-21), which are designated as DV3 and DV1, respectively, have higher CXCR4 binding ability than their L-peptide counterparts. They are partially structured in aqueous solution, displaying a turn-like structure over residues 5-8. The small temperature coefficients of His-6 amide proton for both peptides also suggest the formation of a small hydrophobic pocket centered on His-6. The structural features of DV3 are very similar to the reported solution structure of all-L-peptide vMIP-II (1-10) [M.P. Crump, E. Elisseeva, J. Gong, I. Clark-Lewis, B.D. Sykes, Structure/function of human herpesvirus-8 MIP-II (1-71) and the antagonist N-terminal segment (1-10), FEBS Lett. 489 (2001) 171], which is consistent with the notion that D- and L-enantiomeric peptides can adopt mirror image conformations. The NMR structures of the D-peptides provide a structural basis to understand their mechanism of action and design new peptidomimetic analogs to further explore the structure-activity relationship of D-peptide ligand binding to CXCR4

  5. The CENP-T C-Terminus Is Exclusively Proximal to H3.1 and not to H3.2 or H3.3

    Science.gov (United States)

    Abendroth, Christian; Hofmeister, Antje; Hake, Sandra B.; Kamweru, Paul K.; Miess, Elke; Dornblut, Carsten; Küffner, Isabell; Deng, Wen; Leonhardt, Heinrich; Orthaus, Sandra; Hoischen, Christian; Diekmann, Stephan

    2015-01-01

    The kinetochore proteins assemble onto centromeric chromatin and regulate DNA segregation during cell division. The inner kinetochore proteins bind centromeres while most outer kinetochore proteins assemble at centromeres during mitosis, connecting the complex to microtubules. The centromere–kinetochore complex contains specific nucleosomes and nucleosomal particles. CENP-A replaces canonical H3 in centromeric nucleosomes, defining centromeric chromatin. Next to CENP-A, the CCAN multi-protein complex settles which contains CENP-T/W/S/X. These four proteins are described to form a nucleosomal particle at centromeres. We had found the CENP-T C-terminus and the CENP-S termini next to histone H3.1 but not to CENP-A, suggesting that the Constitutive Centromere-Associated Network (CCAN) bridges a CENP-A- and a H3-containing nucleosome. Here, we show by in vivo FRET that this proximity between CENP-T and H3 is specific for H3.1 but neither for the H3.1 mutants H3.1C96A and H3.1C110A nor for H3.2 or H3.3. We also found CENP-M next to H3.1 but not to these H3.1 mutants. Consistently, we detected CENP-M next to CENP-S. These data elucidate the local molecular neighborhood of CCAN proteins next to a H3.1-containing centromeric nucleosome. They also indicate an exclusive position of H3.1 clearly distinct from H3.2, thus documenting a local, and potentially also functional, difference between H3.1 and H3.2. PMID:25775162

  6. The CENP-T C-Terminus Is Exclusively Proximal to H3.1 and not to H3.2 or H3.3

    Directory of Open Access Journals (Sweden)

    Christian Abendroth

    2015-03-01

    Full Text Available The kinetochore proteins assemble onto centromeric chromatin and regulate DNA segregation during cell division. The inner kinetochore proteins bind centromeres while most outer kinetochore proteins assemble at centromeres during mitosis, connecting the complex to microtubules. The centromere–kinetochore complex contains specific nucleosomes and nucleosomal particles. CENP-A replaces canonical H3 in centromeric nucleosomes, defining centromeric chromatin. Next to CENP-A, the CCAN multi-protein complex settles which contains CENP-T/W/S/X. These four proteins are described to form a nucleosomal particle at centromeres. We had found the CENP-T C-terminus and the CENP-S termini next to histone H3.1 but not to CENP-A, suggesting that the Constitutive Centromere-Associated Network (CCAN bridges a CENP-A- and a H3-containing nucleosome. Here, we show by in vivo FRET that this proximity between CENP-T and H3 is specific for H3.1 but neither for the H3.1 mutants H3.1C96A and H3.1C110A nor for H3.2 or H3.3. We also found CENP-M next to H3.1 but not to these H3.1 mutants. Consistently, we detected CENP-M next to CENP-S. These data elucidate the local molecular neighborhood of CCAN proteins next to a H3.1-containing centromeric nucleosome. They also indicate an exclusive position of H3.1 clearly distinct from H3.2, thus documenting a local, and potentially also functional, difference between H3.1 and H3.2.

  7. The cytoplasmic C-terminus of polycystin-1 increases cell proliferation in kidney epithelial cells through serum-activated and Ca2+-dependent pathway(s)

    International Nuclear Information System (INIS)

    Manzati, Elisa; Aguiari, Gianluca; Banzi, Manuela; Manzati, Michele; Selvatici, Rita; Falzarano, Sofia; Maestri, Iva; Pinton, Paolo; Rizzuto, Rosario; Senno, Laura del

    2005-01-01

    Polycystin-1 (PC1) is a large transmembrane protein important in renal differentiation and defective in most cases of autosomal dominant polycystic kidney disease (ADPKD), a common cause of renal failure in adults. Although the genetic basis of ADPKD has been elucidated, molecular and cellular mechanisms responsible for the dysregulation of epithelial cell growth in ADPKD cysts are still not well defined. We approached this issue by investigating the role of the carboxyl cytoplasmic domain of PC1 involved in signal transduction on the control of kidney cell proliferation. Therefore, we generated human HEK293 cells stably expressing the PC1 cytoplasmic tail as a membrane targeted TrkA-PC1 chimeric receptor protein (TrkPC1). We found that TrkPC1 increased cell proliferation through an increase in cytoplasmic Ca 2+ levels and activation of PKCα, thereby upregulating D1 and D3 cyclin, downregulating p21 waf1 and p27 kip1 cyclin inhibitors, and thus inducing cell cycle progression from G0/G1 to the S phase. Interestingly, TrkPC1-dependent Ca 2+ increase and PKCα activation are not constitutive, but require serum factor(s) as parallel component. In agreement with this observation, a significant increase in ERK1/2 phosphorylation was observed. Consistently, inhibitors specifically blocking either PKCα or ERK1/2 prevented the TrkPC1-dependent proliferation increase. NGF, the TrkA ligand, blocked this increase. We propose that in kidney epithelial cells the overexpression of PC1 C-terminus upregulates serum-evoked intracellular Ca 2+ by counteracting the growth-suppression activity of endogenous PC1 and leading to an increase in cell proliferation

  8. Critical role in CXCR4 signaling and internalization of the polypeptide main chain in the amino terminus of SDF-1α probed by novel N-methylated synthetically and modularly modified chemokine analogues.

    Science.gov (United States)

    Dong, Chang-Zhi; Tian, Shaomin; Choi, Won-Tak; Kumar, Santhosh; Liu, Dongxiang; Xu, Yan; Han, Xiaofeng; Huang, Ziwei; An, Jing

    2012-07-31

    The replication of human immunodeficiency virus type 1 (HIV-1) can be profoundly inhibited by the natural ligands of two major HIV-1 coreceptors, CXCR4 and CCR5. Stromal cell-derived factor-1α (SDF-1α) is a natural ligand of CXCR4. We have recently developed a synthetic biology approach of using synthetically and modularly modified (SMM)-chemokines to dissect various aspects of the structure-function relationship of chemokines and their receptors. Here, we used this approach to design novel SMM-SDF-1α analogues containing unnatural N-methylated residues in the amino terminus to investigate whether the polypeptide main chain amide bonds in the N-terminus of SDF-1α play a role in SDF-1α signaling via CXCR4 and/or receptor internalization. The results show that SDF-1α analogues with a modified N-methylated main chain at position 2, 3, or 5 retain significant CXCR4 binding and yet completely lose signaling activities. Furthermore, a representative N-methylated analogue has been shown to be incapable of causing CXCR4 internalization. These results suggest that the ability of SDF-1α to activate CXCR4 signaling and internalization is dependent upon the main chain amide bonds in the N-terminus of SDF-1α. This study demonstrates the feasibility and value of applying a synthetic biology approach to chemically engineer natural proteins and peptide ligands as probes of important biological functions that are not addressed by other biological techniques.

  9. Innovation for creating a smart future

    Directory of Open Access Journals (Sweden)

    Sang M. Lee

    2018-01-01

    Full Text Available Today, we live in a dynamic and turbulent global community. The wave of mega-trends, including rapid change in globalization and technological advances, is creating new market forces. For any organization to survive and prosper in such an environment, innovation is imperative. However, innovation is no longer just for creating value to benefit individuals, organizations, or societies. The ultimate purpose of innovation should be much more far reaching, helping create a smart future where people can enjoy the best quality of life possible. Thus, innovation must search for intelligent solutions to tackle major social ills, seek more proactive approaches to predict the uncertain future, and pursue strategies to remove barriers to the smart future. This study explores the detailed requirements of a smart future, including both hardware types and soft social/cultural components.

  10. Can Physical Examination Create a Stener Lesion?

    Science.gov (United States)

    Lankachandra, Manesha; Eggers, John P; Bogener, James W; Hutchison, Richard L

    2017-09-01

    The purpose of this study is to determine whether a Stener lesion can be created while testing stability of the ulnar collateral ligament (UCL) of the thumb. Testing was performed in a manner that reproduced clinical examination. Six fresh frozen hand and forearm specimens underwent sequential sectioning of the accessory UCL, the proper UCL, and the ulnar sagittal band. Measurements of radial deviation of the metacarpophalangeal (MCP) joint were taken with the thumb in neutral rotation, pronation and supination, both with 0 degrees and with 30 degrees of flexion of the MCP joint. Visual examination was performed to assess the presence of a Stener lesion. No Stener lesion was created in any position as long as the fascial origin of the ulnar sagittal band at the adductor pollicis longus remained intact. After creating a defect in the ulnar sagittal band, a Stener lesion was created in two specimens, but only when the thumb was flexed and supinated. Pronation provided more stability, and supination provided less stability, with one or both components cut, especially when testing at 30° of flexion. Compared to both components cut without flexion or rotation, there was a statistically significant difference in angulation with the 30 degrees of MCP joint flexion in both neutral rotation in supination. Performing a physical examination to assess the amount of instability of an ulnar collateral ligament injury did not create a Stener lesion if the exam was performed in a controlled, gentle manner with the thumb held without rotation. If the thumb is held in neutral rotation during the exam, an iatrogenic Stener lesion should not be created.

  11. Creating dynamic UI with Android fragments

    CERN Document Server

    Wilson, Jim

    2013-01-01

    A fast-paced tutorial that guides you through everything you need to know about dynamic UI design for Android devices.This book is for developers with a basic understanding of Android programming who would like to improve the appearance and usability of their applications. Whether you're looking to create a more interactive user experience, create more dynamically adaptive UIs, provide better support for tablets and smartphones in a single app, reduce the complexity of managing your app UIs, or you are just trying to expand your UI design philosophy, then this book is for you.

  12. Creating e-learning games with Unity

    CERN Document Server

    Horachek, David

    2014-01-01

    Unity is a fully integrated development engine providing the required functionality to create games and interactive 3D content, while reducing the time, effort, and cost of developing the content. Nowadays, many people have started to use Unity in an eLearning setting as it allows them to create real-world scenarios, or models, for training purposes. With Unity, one can develop video games that are not only fun, but are also effective teaching and learning tools. When properly designed, an engaging game is an ideal platform for the presentation, testing, and application of learning objectives.

  13. Creating a Website The Missing Manual

    CERN Document Server

    MacDonald, Matthew

    2011-01-01

    Think you need an army of skilled programmers to build a website? Think again. With nothing more than an ordinary PC, some raw ambition, and this book, you'll learn how to create and maintain a professional-looking, visitor-friendly site. This Missing Manual gives you all the tools, techniques, and expert advice you need. Plan your site. Create web pages by learning the basics of HTML and HTML5.Control page design with CSS. Format text, images, links, tables, and other elements.Attract visitors. Ensure that people can find your site through popular search engines.Build a community. Add forums

  14. Creating a web site the missing manual

    CERN Document Server

    MacDonald, Matthew

    2008-01-01

    Think you have to be a technical wizard to build a great web site? Think again. If you want to create an engaging web site, this thoroughly revised, completely updated edition of Creating a Web Site: The Missing Manual demystifies the process and provides tools, techniques, and expert guidance for developing a professional and reliable web presence. Whether you want to build a personal web site, an e-commerce site, a blog, or a web site for a specific occasion or promotion, this book gives you detailed instructions and clear-headed advice for: Everything from planning to launching. From pi

  15. Creating library tutorials for nursing students.

    Science.gov (United States)

    Schroeder, Heidi

    2010-04-01

    This article describes one librarian's experiences with creating, promoting, and assessing online library tutorials. Tutorials were designed to provide on-demand and accessible library instruction to nursing students at Michigan State University. Topics for tutorials were chosen based on the librarian's liaison experiences and suggestions from nursing faculty. The tutorials were created using Camtasia and required the application of several tools and techniques. Tutorials were promoted through Web pages, the ANGEL course management system, blog posts, librarian interactions, e-mails, and more. In order to assess the tutorials' perceived effectiveness, feedback was gathered using a short survey. Future plans for the nursing tutorials project are also discussed.

  16. How to create a serious game?

    Directory of Open Access Journals (Sweden)

    O. Heidmann

    2015-11-01

    Full Text Available Serious games are video games designed to achieve an educational effect and achieve some degree of training in a certain area. They are nowadays used in industries such as defense, education, scientific exploration, health care, emergency management, city planning, engineering, and many others. As it still a nascent subject who doesn’t follow exactly the same rules and practices than the commercial video games industry, questions remain about how to create and use serious games. This article presents some know-how on the subject of creating serious games.

  17. Creating a Total Object of Art.

    Science.gov (United States)

    Klie, Evelyn Busch

    2003-01-01

    Discusses a workshop that accompanied the author's exhibit, "A Sense of Place: Paintings by Evelyn Busch Klie." Explains that students created a watercolor painting and a clay frame or base with details in it. Includes a list of art materials and learning objectives. (CMK)

  18. Strategies for Creating New Venture Legitimacy

    Science.gov (United States)

    Karlsson, Tomas; Middleton, Karen Williams

    2015-01-01

    New ventures, being heavily subjected to liabilities of newness, are seen to engage in legitimacy strategies to overcome these liabilities. Building on an adapted theoretical framework of organizational legitimacy, self-reported weekly diaries of twelve entrepreneurs were analysed to identify strategies used by new ventures to create legitimacy.…

  19. Creating by Reusing Learning Design Solutions

    NARCIS (Netherlands)

    Hernández-Leo, Davinia; Harrer, Andreas; Dodero, Juan Manuel; Asensio-Pérez, Juan; Burgos, Daniel

    2006-01-01

    Hernández-Leo, D., Harrer, A., Dodero, J. M., Asension-Pérez, J. I., & Burgos, D. (2006). Creating by reusing Learning Design solutions. Proceedings of 8th Simposo Internacional de Informática Educativa, León, Spain: IEEE Technical Committee on Learning Technology. Retrieved October 3rd, 2006, from

  20. Creating Innovative Student Projects with App Smashing

    Science.gov (United States)

    Young, Donna

    2014-01-01

    The potential for using various apps to improve student learning is tremendous. Yet, despite the iPad's possibilities, apps are often limited in their functionality. No one has created that magical, one-size-fits-all app that accomplishes all of the tasks that you had in mind. Luckily, there is an answer to this common problem: app smashing.…

  1. How do entrepreneurs think they create value?

    DEFF Research Database (Denmark)

    Lyth Frederiksen, Dennis; Brem, Alexander

    2017-01-01

    The means with which entrepreneurs create and capture value can be difficult to get a comprehensive picture of. Looking at the tools they use can offer insights, and in this context, the book “The Lean Startup” by Eric Ries has received a tremendous amount of attention. Supposedly, many...

  2. Creating social impact with side-events

    NARCIS (Netherlands)

    Hover, Paul; Dijk, Bake; Breedveld, Koen; van Eekeren, Frank; Slender, Hans

    All over the world, sport events are seen as significant tools for creating positive social impact. This is understandable, as sport events have the power to attract enthusiastic participants, volunteers and to reach large audiences of visitors and followers via (social) media. Outbursts of

  3. Creating Spaces to Support Transgender Youth

    Science.gov (United States)

    McGuire, Jenifer K.; Conover-Williams, Meredith

    2010-01-01

    This article explores the opportunity to create spaces within the family, school, and community that specifically promote the well-being of transgender adolescents and young adults. When social contexts are supportive, transgender youth report significantly less risk. Supportive home and school environments have been linked to better outcomes…

  4. Does the New Economy Create Higher Productivity?

    DEFF Research Database (Denmark)

    Dilling-Hansen, Mogens; Madsen, Erik Strøjer; Smith, Valdemar

    2002-01-01

    The rapid and continuous growth in the US in the 1990s and the simultaneous boom in the IT industry created the concept "The New Economy". What connects the two phenomena is that the IT industry alone is considered productive, and increased productivity in other industries, as a result of increased...

  5. The NAi effect: creating architecture culture

    NARCIS (Netherlands)

    Figueiredo, S.M.

    2016-01-01

    At a time when cultural institutions (both in the Netherlands and abroad) are adjusting to new societal and political conditions (and financial restrictions), The NAi Effect aims to elucidate the social and cultural aims of architecture museums and their impact in creating architecture culture.

  6. Creating a Sun-Safe Camp.

    Science.gov (United States)

    Landrey, Ann

    1996-01-01

    Strategies for minimizing sun exposure of campers and staff include educating campers about the sun's effect on their skin, scheduling activities when the sun is less intense, creating shade at the camp site, incorporating sun protection into camp dress code, and training staff regarding sun protection. Addresses OSHA and liability issues. (LP)

  7. Inclusive Information Societies: Creating Growth and Employment ...

    International Development Research Centre (IDRC) Digital Library (Canada)

    Employment for youth and women. The project has two components. The first focuses on outsourcing service sector work (micro-work and rural impact sourcing) in India, Sri Lanka, and the Philippines. This sector can play a vital role in creating employment, particularly for youth and women in semi-urban and rural areas.

  8. Creating the networking enterprises - logistics determinants

    Directory of Open Access Journals (Sweden)

    Ewa Kulińska

    2014-06-01

    Full Text Available Background: The article describes the determinants of creating network enterprises with peculiar consideration of logistic factors which are conditioning the organization of processes, exchange of resources and competences. On the basis of literature analysis, there is proposed a model of creating network enterprises. A model is verified in the application part of the thesis. Methods: Within the publication a literature review of submitted scope of the interest was presented, as well as the empirical research. A research substance attaches the enterprises created on the basis of the reactivation of organizations which has collapsed due to bankruptcy proceeding. The research was based upon direct interviews with employees of the net-forming entities. Results and conclusions: Results of the research shows that taking up the cooperation and net-cooperation was the only possibility for new entities to come into existence, that were  based upon old assets and human resources liquidated during bankruptcy proceeding. There was indentified many determinants of enterprises network cooperation, however due to the research a conclusion draws, that basic factors of creating network cooperation are those which are profit-achieving oriented.

  9. CREATING INPUT TABLES FROM WAPDEG FOR RIP

    International Nuclear Information System (INIS)

    K.G. Mon

    1998-01-01

    The purpose of this calculation is to create tables for input into RIP ver. 5.18 (Integrated Probabilistic Simulator for Environmental Systems) from WAPDEG ver. 3.06 (Waste Package Degradation) output. This calculation details the creation of the RIP input tables for TSPA-VA REV.00

  10. Leadership: creating a cuiture of caring.

    Science.gov (United States)

    DePaola, Dominick P

    2004-01-01

    Leadership is characterized in terms of accomplishing mutual goals for the organization, its employees, and its community through vision and creating a community of caring. The examples of Herb Kelleher of Southwest Airlines, Walt Disney, and Dean Arthur A. Dugoni of the University of the Pacific are used to illustrate how this style of leadership plays out in specific accomplishments.

  11. System and method for creating expert systems

    Science.gov (United States)

    Hughes, Peter M. (Inventor); Luczak, Edward C. (Inventor)

    1998-01-01

    A system and method provides for the creation of a highly graphical expert system without the need for programming in code. An expert system is created by initially building a data interface, defining appropriate Mission, User-Defined, Inferred, and externally-generated GenSAA (EGG) data variables whose data values will be updated and input into the expert system. Next, rules of the expert system are created by building appropriate conditions of the rules which must be satisfied and then by building appropriate actions of rules which are to be executed upon corresponding conditions being satisfied. Finally, an appropriate user interface is built which can be highly graphical in nature and which can include appropriate message display and/or modification of display characteristics of a graphical display object, to visually alert a user of the expert system of varying data values, upon conditions of a created rule being satisfied. The data interface building, rule building, and user interface building are done in an efficient manner and can be created without the need for programming in code.

  12. Practice It: Create a Weekly Activity Plan

    Science.gov (United States)

    At the beginning of the week, create an activity plan to help you reach your goals. Start by identifying your goals for the week. Based on your goals, write down when you are going to exercise and what you are going to do.

  13. Understanding Critical Thinking to Create Better Doctors

    Science.gov (United States)

    Zayapragassarazan, Zayabalaradjane; Menon, Vikas; Kar, Sitanshu Sekhar; Batmanabane, Gitanjali

    2016-01-01

    Medical students master an enormous body of knowledge, but lack systematic problem solving ability and effective clinical decision making. High profile reports have called for reforms in medical education to create a better generation of doctors who can cope with the system based problems they would encounter in an interdisciplinary and…

  14. Creating an Economically Enabling and Competitive Business ...

    International Development Research Centre (IDRC) Digital Library (Canada)

    Creating an Economically Enabling and Competitive Business Environment in the West Bank and Gaza Strip. The prospect of indefinite Israeli occupation of the Palestinian territories, and their extreme dependence on foreign assistance and Israeli-controlled customs revenues, had led to the conclusion that the Palestinian ...

  15. Creating an Economically Enabling and Competitive Business ...

    International Development Research Centre (IDRC) Digital Library (Canada)

    Creating an Economically Enabling and Competitive Business Environment in the West Bank and Gaza Strip. The prospect of indefinite Israeli occupation of the ... Impact of implementing the Palestinian banking law on the performance of the private sector [Arabic language]. Documents. Impact of the commercial agents law ...

  16. Creating Smart-er Cities: An Overview

    Science.gov (United States)

    Allwinkle, Sam; Cruickshank, Peter

    2011-01-01

    The following offers an overview of what it means for cities to be "smart." It draws the supporting definitions and critical insights into smart cities from a series of papers presented at the 2009 Trans-national Conference on Creating Smart(er) Cities. What the papers all have in common is their desire to overcome the all too often…

  17. Engineering Encounters: Creating a Prosthetic Hand

    Science.gov (United States)

    Cook, Kristin Leigh; Bush, Sarah B.; Cox, Richard

    2015-01-01

    The power of 3D printing technology has grown exponentially in just the past few years--people around the world are using 3D printers to prepare food, create tailored clothing, build cars and homes, and advance the medical field in ways that never seemed possible. In classrooms across the nation, 3D printers have become increasingly common because…

  18. Photocatalytic Solutions Create Self-Cleaning Surfaces

    Science.gov (United States)

    2013-01-01

    A Stennis Space Center researcher investigating the effectiveness of photocatalytic materials for keeping the Center's buildings free of grime turned to a solution created by PURETi Inc. of New York City. Testing proved successful, and NASA and the company now share a Dual Use Technology partnership. PURETi's coatings keep surfaces clean and purify surrounding air, eliminating pollution, odors, and microbes.

  19. Creating Sustainable Development through Sustainability Bildung

    African Journals Online (AJOL)

    2018-05-12

    May 12, 2018 ... ... to quit their jobs, sell their house, take their children out of school ... the globe is perceived as an object that human beings can affect, damage or protect, but ... have created a fantasy in which we believe that we can distance ...

  20. Creating Teams Increases Extension Educator Productivity

    Science.gov (United States)

    Chalker-Scott, Linda; Daniels, Catherine H.; Martini, Nicole

    2016-01-01

    The Garden Team at Washington State University is a transdisciplinary group of faculty, staff, and students with expertise in applied plant and soil sciences and an interest in Extension education. The team's primary mission is to create current, relevant, and peer-reviewed materials as Extension publications for home gardeners. The average yearly…

  1. creating social presence in large classes

    African Journals Online (AJOL)

    Social presence refers to the ability of students to project themselves as 'real people' in an online learning community. While it is difficult to create social presence in large classes, educational technologies can enhance the social dimension of online learning if educators relinquish the use of technology as an instrument of ...

  2. The Media Creates Us in Its Image

    Science.gov (United States)

    Stivers, Richard

    2012-01-01

    Propaganda in all its forms is the culture of a mass society. The media transmits propaganda to form public opinion and recreate the human being. Reversing the Western ideal of a rational and free individual, the media creates a childish conformist ensconced in the peer group, who acts unconsciously.

  3. Creating a winning team: lessons from football.

    Science.gov (United States)

    Simons, Sherri Lee

    2005-01-01

    There are tasks best done on an individual basis when caring for a neonate, but the ultimate outcome for infants and their families results from a team effort. Incorporating ten strategies drawn from football can help the NICU manager create and foster effective teamwork.

  4. Creating a Data Warehouse using SQL Server

    DEFF Research Database (Denmark)

    Sørensen, Jens Otto; Alnor, Karl

    1999-01-01

    In this paper we construct a Star Join Schema and show how this schema can be created using the basic tools delivered with SQL Server 7.0. Major objectives are to keep the operational database unchanged so that data loading can be done with out disturbing the business logic of the operational...

  5. Creating social impact with sport events

    NARCIS (Netherlands)

    Hover, P.; Dijk, B.; Breedveld, K.; Eekeren, F.J.A. van; Slender, H.

    2016-01-01

    All over the world, sport events are seen as significant tools for creating positive social impact. This is understandable, as sport events have the power to attract enthusiastic participants, volunteers and to reach large audiences of visitors and followers via (social) media. Outbursts of

  6. Creating the Grateful School in Four Phases

    Science.gov (United States)

    Griffith, Owen M.

    2018-01-01

    Author and educator Owen Griffith shares how leaders can infuse small acts of gratitude to energize their schools and create a positive, thriving culture. Leaders must begin by practicing gratitude personally, then slowly introducing the practice to their faculty and eventually students. With the right attitude and creativity, the acts of…

  7. Creating the Total Quality Effective School.

    Science.gov (United States)

    Lezotte, Lawrence W.

    This book shows how Deming's Total Quality Management (TQM) theory for organizational management can be integrated with the effective-schools literature. Part 1 compares the 14 principles of TQM with the tenets of effective-schools research. The second part develops a blueprint for creating the total quality effective school. The conceptual…

  8. Extending the "Knowledge Advantage": Creating Learning Chains

    Science.gov (United States)

    Maqsood, Tayyab; Walker, Derek; Finegan, Andrew

    2007-01-01

    Purpose: The purpose of this paper is to develop a synergy between the approaches of knowledge management in a learning organisation and supply chain management so that learning chains can be created in order to unleash innovation and creativity by managing knowledge in supply chains. Design/methodology/approach: Through extensive literature…

  9. Creating science simulations through Computational Thinking Patterns

    Science.gov (United States)

    Basawapatna, Ashok Ram

    Computational thinking aims to outline fundamental skills from computer science that everyone should learn. As currently defined, with help from the National Science Foundation (NSF), these skills include problem formulation, logically organizing data, automating solutions through algorithmic thinking, and representing data through abstraction. One aim of the NSF is to integrate these and other computational thinking concepts into the classroom. End-user programming tools offer a unique opportunity to accomplish this goal. An end-user programming tool that allows students with little or no prior experience the ability to create simulations based on phenomena they see in-class could be a first step towards meeting most, if not all, of the above computational thinking goals. This thesis describes the creation, implementation and initial testing of a programming tool, called the Simulation Creation Toolkit, with which users apply high-level agent interactions called Computational Thinking Patterns (CTPs) to create simulations. Employing Computational Thinking Patterns obviates lower behavior-level programming and allows users to directly create agent interactions in a simulation by making an analogy with real world phenomena they are trying to represent. Data collected from 21 sixth grade students with no prior programming experience and 45 seventh grade students with minimal programming experience indicates that this is an effective first step towards enabling students to create simulations in the classroom environment. Furthermore, an analogical reasoning study that looked at how users might apply patterns to create simulations from high- level descriptions with little guidance shows promising results. These initial results indicate that the high level strategy employed by the Simulation Creation Toolkit is a promising strategy towards incorporating Computational Thinking concepts in the classroom environment.

  10. Genetic diversity in the C-terminus of merozoite surface protein 1 among Plasmodium knowlesi isolates from Selangor and Sabah Borneo, Malaysia.

    Science.gov (United States)

    Yap, Nan Jiun; Goh, Xiang Ting; Koehler, Anson V; William, Timothy; Yeo, Tsin Wen; Vythilingam, Indra; Gasser, Robin B; Lim, Yvonne A L

    2017-10-01

    Plasmodium knowlesi, a malaria parasite of macaques, has emerged as an important parasite of humans. Despite the significance of P. knowlesi malaria in parts of Southeast Asia, very little is known about the genetic variation in this parasite. Our aim here was to explore sequence variation in a molecule called the 42kDa merozoite surface protein-1 (MSP-1), which is found on the surface of blood stages of Plasmodium spp. and plays a key role in erythrocyte invasion. Several studies of P. falciparum have reported that the C-terminus (a 42kDa fragment) of merozoite surface protein-1 (MSP-1 42 ; consisting of MSP-1 19 and MSP-1 33 ) is a potential candidate for a malaria vaccine. However, to date, no study has yet investigated the sequence diversity of the gene encoding P. knowlesi MSP-1 42 (comprising Pk-msp-1 19 and Pk-msp-1 33 ) among isolates in Malaysia. The present study explored this aspect. Twelve P. knowlesi isolates were collected from patients from hospitals in Selangor and Sabah Borneo, Malaysia, between 2012 and 2014. The Pk-msp-1 42 gene was amplified by PCR and directly sequenced. Haplotype diversity (Hd) and nucleotide diversity (л) were studied among the isolates. There was relatively high genetic variation among P. knowlesi isolates; overall Hd and л were 1±0.034 and 0.01132±0.00124, respectively. A total of nine different haplotypes related to amino acid alterations at 13 positions, and the Pk-MSP-1 19 sequence was found to be more conserved than Pk-msp-1 33 . We have found evidence for negative selection in Pk-msp- 42 as well as the 33kDa and 19kDa fragments by comparing the rate of non-synonymous versus synonymous substitutions. Future investigations should study large numbers of samples from disparate geographical locations to critically assess whether this molecule might be a potential vaccine target for P. knowlesi. Copyright © 2017 Elsevier B.V. All rights reserved.

  11. PROTEOLYTIC REMOVAL OF THE CARBOXYL TERMINUS OF THE T4 GENE 32 HELIX-DESTABILIZING PROTEIN ALTERS THE T4 IN VITRO REPLICATION COMPLEX

    Energy Technology Data Exchange (ETDEWEB)

    Burke, R.L.; Alberts, B.M.; Hosoda, J.

    1980-07-01

    The proteolytic removal of about 60 amino acids from the COOH terminus of the bacteriophage T4 helix-destabilizing protein (gene 32 protein) produces 32*I, a 27,000-dalton fragment which still binds tightly and cooperatively to single-stranded DNA. The substitution of 32*I protein for intact 32 protein in the seven-protein T4 replication complex results in dramatic changes in some of the reactions catalyzed by this in vitro DNA replication system, while leaving others largely unperturbed. (1) Like intact 32 protein, the 32*I protein promotes DNA synthesis by the DNA polymerase when the T4 polymerase accessory proteins (gene 44/62 and 45 proteins) are also present. The host helix-destabilizing protein (Escherichia coli ssb protein) cannot replace the 32*I protein for this synthesis. (2) Unlike intact 32 protein, 32*I protein strongly inhibits DNA synthesis catalyzed by the T4 DNA polymerase alone on a primed single-stranded DNA template. (3) Unlike intact 32 protein, the 32*I protein strongly inhibits RNA primer synthesis catalyzed by the T4 gene 41 and 61 proteins and also reduces the efficiency of RNA primer utilization. As a result, de novo DNA chain starts are blocked completely in the complete T4 replication system, and no lagging strand DNA synthesis occurs. (4) The 32*I protein does not bind to either the T4 DNA polymerase or to the T4 gene 61 protein in the absence of DNA; these associations (detected with intact 32 protein) would therefore appear to be essential for the normal control of 32 protein activity, and to account at least in part for observations 2 and 3, above. We propose that the COOH-terminal domain of intact 32 protein functions to guide its interactions with the T4 DNA polymerase and the T4 gene 61 RNA-priming protein. When this domain is removed, as in 32*I protein, the helix destabilization induced by the protein is controlled inadequately, so that polymerizing enzymes tend to be displaced from the growing 3{prime}-OH end of a

  12. Translation vs. Rotation: The Battle for Accommodation of Dextral Shear at the Northern Terminus of the Central Walker Lane, Western Nevada

    Science.gov (United States)

    Carlson, C. W.; Faulds, J. E.

    2014-12-01

    Positioned between the Sierra Nevada microplate and Basin and Range in western North America, the Walker Lane (WL) accommodates ~20% of the dextral motion between the North American and Pacific plates on predominately NW-striking dextral and ENE to E-W-striking sinistral fault systems. The Terrill Mountains (TM) lie at the northern terminus of a domain of dextral faults accommodating translation of crustal-blocks in the central WL and at the southeast edge of sinistral faults accommodating oroclinal flexure and CW rotation of blocks in the northern WL. As the mechanisms of strain transfer between these disparate fault systems are poorly understood, the thick Oligocene to Pliocene volcanic strata of the TM area make it an ideal site for studying the transfer of strain between regions undergoing differing styles of deformation and yet both accommodating dextral shear. Detailed geologic mapping and paleomagnetic study of ash-flow tuffs in the TM region has been conducted to elucidate Neogene strain accommodation for this transitional region of the WL. Strain at the northernmost TM appears to be transferred from a system of NW-striking dextral faults to a system of ~E-W striking sinistral faults with associated CW flexure. A distinct ~23 Ma paleosol is locally preserved below the tuff of Toiyabe and provides an important marker bed. This paleosol is offset with ~6 km of dextral separation across the fault bounding the NE flank of the TM. This fault is inferred as the northernmost strand of the NW-striking, dextral Benton Spring fault system, with offset consistent with minimums constrained to the south (6.4-9.6 km, Gabbs Valley Range). Paleomagnetic results suggest counter-intuitive CCW vertical-axis rotation of crustal blocks south of the domain boundary in the system of NW-striking dextral faults, similar to some other domains of NW-striking dextral faults in the northern WL. This may result from coeval dextral shear and WNW-directed extension within the left

  13. Creating a culture for information systems success

    CERN Document Server

    Belkhamza, Zakariya

    2015-01-01

    It has been widely reported that issues related to organizational context appear frequently in discussions of information systems success. The statement that the information system did not fit the behavioral context in an organization is often part of the explanation of why particular information system encountered unanticipated resistance and never met expectation. While this context has been intensively studied, we still lack evidence on how this organizational context is affecting the success of information system from a managerial action perspective. This type of managerial involvement is often neglected to the extent that it became an essential obstacle to organizational performance. The objective of Creating a Culture for Information Systems Success is to assist CIOs and IT managers on how to use their managerial actions to create a suitable cultural environment in the organization, which leads to a successful implementation of information systems. This  book will also provide guidelines fo...

  14. Designing value-creating supply chain networks

    CERN Document Server

    Martel, Alain

    2016-01-01

    Focusing on the design of robust value-creating supply chain networks (SCN) and key strategic issues related to the number; location, capacity and mission of supply chain facilities (plants, distribution centers) – as well as the network structure required to provide flexibility and resilience in an uncertain world – this book presents an innovative methodology for SCN reengineering that can be used to significantly improve the bottom line of supply chain dependent businesses. Providing readers with the tools needed to analyze and model value creation activities, Designing Value-Creating Supply Chain Networks examines the risks faced by modern supply chains, and shows how to develop plausible future scenarios to evaluate potential SCN designs. The design methods proposed are based on a visual representation formalism that facilitates the analysis and modeling of SCN design problems, book chapters incorporate several example problems and exercises which can be solved with Excel tools (Analysis tools and So...

  15. Creating sustainable city by enhancing social capital

    Science.gov (United States)

    Affandi, R. A.; Mursitama, T. N.

    2018-03-01

    Scholars have been discussing social capital since the last two decades. They analyzed from various perspectives such as sociology, education, political participation, strengthening democratic values and economic empowerment of the society. However, study related to the implementation that benefits directly to the society is needed. This study examines how to create a sustainable city by enhancing social capital from both macro and micro analyses. This combination of analysis offers deeper understanding both from decision makers at city level and individuals, groups and society. We will conduct qualitative approach mainly by interviews and direct observation to collect the data. also, we also analyze publicly available data. Finally, this study contributes to new understanding in creating a sustainable city, not only about the environment and physical aspects, but also about ensuring political economic, democratic values, and social welfare.

  16. Digital Media Creates Youth Voices Heard

    Directory of Open Access Journals (Sweden)

    Jeff Sallee

    2014-06-01

    Full Text Available Oklahoma 4-H clubs and military service centers partnered with the Adobe Youth Voices (AYV program to give youth opportunities to raise their voices through digital media. This program reached out to underrepresented youth and gave them the tools and technology to effectively express themselves. The intent of this project was for 4-H members to create videos to educate, help and raise awareness in their communities of topics that were important to the youth. These experiences help youth gain knowledge towards helping others solve farm, home, and community problems. Participating youth selected issues that were important to them and created a short video, educating others and sharing their convictions on the topics of horse therapy, citizenship, bullying, and distracted driving.

  17. Co-creating meaning through Artful Inquiry

    DEFF Research Database (Denmark)

    Darsø, Lotte

    2017-01-01

    The purpose of this chapter is to point out the need for aesthetic and artful methods for reflection, learning and co-creation. The context is management education focused on developing innovation competency. The data derive from action research, observations and written reports. The main contrib...... of leadership icons as well as co-creating with tangible materials can give rise to new meaning and transformational learning.......The purpose of this chapter is to point out the need for aesthetic and artful methods for reflection, learning and co-creation. The context is management education focused on developing innovation competency. The data derive from action research, observations and written reports. The main...... contribution of this chapter is the introduction of a model for Artful Inquiry, which involves constructing powerful questions and finding appropriate artistic methods for reflecting and for co-creating with people or with artistic material. It is argued that Artful Inquiry can access deeper layers of knowing...

  18. Creating interdisciplinary education within monodisciplinary structures

    DEFF Research Database (Denmark)

    Lindvig, Katrine; Lyall, Catherine; R. Meagher, Laura

    2017-01-01

    The literature on interdisciplinary higher education is influenced by two overall trends: one looks at the institutional level of specially designed interdisciplinary institutions, while the other assesses individual interdisciplinary educational activities. Much less attention is given...... to the processes of creating interdisciplinary education initiatives within traditional monodisciplinary universities. In this study, we thus explore how interdisciplinary education and teaching emerge and develop within universities that have little or no established infrastructure to support interdisciplinarity....... Using qualitative data from a multi-part case study, we examine the development of diverse interdisciplinary educational efforts within a traditional faculty-structured university in order to map the ways in which interdisciplinary educational elements have been created, supported, challenged or even...

  19. Novel device for creating continuous curvilinear capsulorhexis.

    Science.gov (United States)

    Soylak, Mustafa

    2016-01-01

    The purpose of this paper is to develop a novel capsulorhexis system. Mechatronics Laboratory, University of Erciyes and Kayseri Maya Eye Hospital. A 3D model was created and simulations were conducted to develop a new device which was designed, fabricated and tested for continuous curvilinear capsulorhexis (CCC). The name of this system is the electro-mechanical capsulorhexis system (EMCS). The 3D model was created by using a commercial design software and a 3D printer was used to fabricate the EMCS Finite element analysis and geometrical relation tests of the EMCS for different sized lenses were performed. The results show that the EMCS is a perfect solution for capsulorhexis surgeries, without mechanical or geometrical problems. The EMCS can open the anterior lens capsule more easily and effectively than manual CCC applications and needs less experience.

  20. Can cognitive science create a cognitive economics?

    Science.gov (United States)

    Chater, Nick

    2015-02-01

    Cognitive science can intersect with economics in at least three productive ways: by providing richer models of individual behaviour for use in economic analysis; by drawing from economic theory in order to model distributed cognition; and jointly to create more powerful 'rational' models of cognitive processes and social interaction. There is the prospect of moving from behavioural economics to a genuinely cognitive economics. Copyright © 2014. Published by Elsevier B.V.

  1. Story-dialogue: creating community through storytelling

    OpenAIRE

    Doyle-Jones, Carol Sarah

    2006-01-01

    This narrative case study examines the role of storytelling in creating community with a grade 7 class. Twelve girls and eleven boys, ages 12 to 13, participated in this classroom-based study. Students engaged in three structured storytelling activities incorporating home-to-school stories, story responses, and classroom presentations. First, students’ parents/guardians told a coming-of-age or Confirmation story to their child. Second, at school, students shared their family story with a part...

  2. Strong branding creates a competitive edge.

    Science.gov (United States)

    Wagner, Peggy

    2007-01-01

    This article encompasses the basics of branding and how it relates to radiology organizations. It also provides tools to help develop your brand. To effectively use branding as a component of your marketing strategy, it is important to follow 3 basic principles: focus on where you excel, understand the existing markets, and be consistent. You do not need to be a large hospital, imaging center, or department to create a brand identity.

  3. Creating an Asthma-Friendly School

    Centers for Disease Control (CDC) Podcasts

    2007-11-08

    This podcast features real-life success stories of students with asthma who, thanks to their schools' implementation of asthma-friendly policies and programs, now have their asthma under control.  Created: 11/8/2007 by National Center for Chronic Disease Prevention and Health Promotion (NCCDPHP), Division of Adolescent and School Health (DASH).   Date Released: 5/20/2008.

  4. Creating marketing strategies for higher education institutions

    OpenAIRE

    Lidia Białoń

    2015-01-01

    The article presents a thesis that the primary premise of creating marketing strategies for higher education institution is a three-dimensional notion of marketing. The first dimension lies in the theoretical notions of the essence of marketing, including the transactional marketing (1.0), relationship marketing (2.0) and spiritual marketing (3.0). The second dimension is formed by methods of marketing research and accurate notions of marketing, while the third are channels of marketing infor...

  5. Cognitive abilities and creating metaphorical names

    Directory of Open Access Journals (Sweden)

    Avanesyan, Marina O.

    2016-06-01

    Full Text Available The cognitive processing of metaphor creation has been insufficiently investigated. Creating metaphors requires the ability to work in a fantastic, impossible context, using symbolic and associative means to express oneís thoughts. It has been shown recently that intelligence plays an important role in the creation of metaphors, but it is not the main factor in determining their success. The present research explores the roles of conceptual abilities, categorical abilities, and flexibility (as the factor creativity in metaphor creation. Participants (n = 38 young adults were asked to come up with names for three photos, without any special instruction to create metaphors. To classify conceptual abilities we used ìConceptual Synthesisî (M. A. Kholodnaya, 2012; to measure categorical ability we used the subtest ìSimilaritiesî (D. Wechsler, 1955; to identify the role of creativity in the metaphor process we used the test of ìUnusual Usesî (J. P. Guilford, 1960. The creation of complex metaphorical names was associated with a tendency to create highly organized mental structures and to retain them within the general semantic context (r = 0.344, p < 0.05. The tendency to create single-level situational connections was associated with a tendency to give specific names to photos (r = 0.475, p < 0.01. Photographic images proved out to be fruitful stimuli to investigate the processing of visual information. We developed a preliminary classification of names: 1 concrete; 2 situational; 3 abstract; 4 metaphorical (M1 and M2. We identified two types of metaphorical names — perceptual and complex metaphors — that relate to conceptual abilities in different ways. It is inaccurate to speak about a general concept of ìmetaphorical abilitiesî; we should differentiate the psychological mechanisms that lie at their base.

  6. Creating load for new hydrogen production

    International Nuclear Information System (INIS)

    Smith, R.

    2006-01-01

    This presentation provides an update of the activities of the Hydrogen Village. The Hydrogen Village is a public-private partnership of approximately 40 companies with the goal of advancing awareness of the environmental, economic and social benefits of hydrogen and fuel cell technologies. The intent of the hydrogen village is to create a sustainable commercial market for these technologies within the Greater Toronto Area and to help to catalyze such markets in other areas

  7. Creating the next generation control system software

    International Nuclear Information System (INIS)

    Schultz, D.E.

    1989-01-01

    A new 1980's style support package for future accelerator control systems is proposed. It provides a way to create accelerator applications software without traditional programming. Visual Interactive Applications (VIA) is designed to meet the needs of expanded accelerator complexes in a more cost effective way than past experience with procedural languages by using technology from the personal computer and artificial intelligence communities. 4 refs

  8. Creating Life in an Urban Space

    OpenAIRE

    Fredrickson, Kirsten I.

    1999-01-01

    Creating Life in an Urban Space Kirsten Fredrickson Abstract Towns contain spaces defined by human interaction with their surroundings. In any town, certain places seem inviting while others seem cold and unfriendly. This is the result of subtle design decisions that directly effect the character of a place. This investigation focuses on the interaction of architecture in our daily lives and how it affects us in ways that we often overlook. The life of a town is in its rela...

  9. Insulin receptor substrate proteins create a link between the tyrosine phosphorylation cascade and the Ca2+-ATPases in muscle and heart.

    Science.gov (United States)

    Algenstaedt, P; Antonetti, D A; Yaffe, M B; Kahn, C R

    1997-09-19

    Following phosphorylation by the insulin receptor kinase, the insulin receptor substrates (IRS)-1 and IRS-2 bind to and activate several Src homology 2 (SH2) domain proteins. To identify novel proteins that interact with IRS proteins in muscle, a human skeletal muscle cDNA expression library was created in the lambdaEXlox system and probed with baculovirus-produced and tyrosine-phosphorylated human IRS-1. One clone of the 10 clones which was positive through three rounds of screening represented the C terminus of the human homologue of the adult fast twitch skeletal muscle Ca2+-ATPase (SERCA1) including the cytoplasmic tail and part of transmembrane region 10. Western blot analysis of extracts of rat muscle demonstrated co-immunoprecipitation of both IRS-1 and IRS-2 with the skeletal muscle Ca2+-ATPase (SERCA1) and the cardiac muscle isoform (SERCA2). In both cases, injection of insulin stimulated a 2- to 6-fold increase in association of which was maximal within 5 min. In primary cultures of aortic smooth muscle cells and C2C12 cells, the insulin-stimulated interaction between IRS proteins and SERCA1 and -2 was dose-dependent with a maximum induction at 100 nM insulin. This interaction was confirmed in a "pull down" experiment using a glutathione S-transferase fusion protein containing the C terminus of the human SERCA isoform and phosphorylated IRS-1 in vitro and could be blocked by a FLVRES-like domain peptide present in the human SERCA sequence. Affinity chromatography of phosphopeptide libraries using the glutathione S-transferase fusion protein of the C terminus of SERCA1 indicated a consensus sequence for binding of XpYGSS; this is identical to potential tyrosine phosphorylation sites at position 431 of human IRS-1 and at position 500 of human IRS-2. In streptozotocin diabetic rats the interaction between IRS proteins and SERCA1 in skeletal muscle and SERCA2 in cardiac muscle was significantly reduced. Taken together, these results indicate that the IRS

  10. CREATE-IP and CREATE-V: Data and Services Update

    Science.gov (United States)

    Carriere, L.; Potter, G. L.; Hertz, J.; Peters, J.; Maxwell, T. P.; Strong, S.; Shute, J.; Shen, Y.; Duffy, D.

    2017-12-01

    The NASA Center for Climate Simulation (NCCS) at the Goddard Space Flight Center and the Earth System Grid Federation (ESGF) are working together to build a uniform environment for the comparative study and use of a group of reanalysis datasets of particular importance to the research community. This effort is called the Collaborative REAnalysis Technical Environment (CREATE) and it contains two components: the CREATE-Intercomparison Project (CREATE-IP) and CREATE-V. This year's efforts included generating and publishing an atmospheric reanalysis ensemble mean and spread and improving the analytics available through CREATE-V. Related activities included adding access to subsets of the reanalysis data through ArcGIS and expanding the visualization tool to GMAO forecast data. This poster will present the access mechanisms to this data and use cases including example Jupyter Notebook code. The reanalysis ensemble was generated using two methods, first using standard Python tools for regridding, extracting levels and creating the ensemble mean and spread on a virtual server in the NCCS environment. The second was using a new analytics software suite, the Earth Data Analytics Services (EDAS), coupled with a high-performance Data Analytics and Storage System (DASS) developed at the NCCS. Results were compared to validate the EDAS methodologies, and the results, including time to process, will be presented. The ensemble includes selected 6 hourly and monthly variables, regridded to 1.25 degrees, with 24 common levels used for the 3D variables. Use cases for the new data and services will be presented, including the use of EDAS for the backend analytics on CREATE-V, the use of the GMAO forecast aerosol and cloud data in CREATE-V, and the ability to connect CREATE-V data to NCCS ArcGIS services.

  11. Molecular and structural characterisation of the human sodium/iodide symporter (h N.I.S.) C-terminus and the implication of this domain in the transporter regulation

    International Nuclear Information System (INIS)

    Huc, S.

    2007-12-01

    The human natrium iodide symporter (h N.I.S.) is an intrinsic membrane protein expressed in thyroid cells where it allows iodide uptake and accumulation. It is composed of thirteen transmembrane helices and its ninety- three amino acids long cytosolic C-terminus presents many potential post-translational regulatory sites. A first part of the PhD work has been dedicated to the expression in a bacterial system and to the purification of the cytosolic C-terminal fragment. Biochemical and structural characterisation have revealed that this C-terminus is very flexible but prone to dimerization. The fragment has also been used as a bait to test the interactions with PDZ domain proteins spotted on a membrane. Several proteins interacting with the (natrium/iodide symporter) N.I.S. C-terminus have thus been identified and the study of their implication in the protein regulation has been initiated. A second part of the work has underlined the existence of a N.I.S. fragment co-purified with the entire protein. This fragment has been found in cells in culture stably expressing N.I.S. and also in human thyroid extracts and in rodent thyroid cells. We observed that this fragment is spontaneously associated with the entire protein. It is composed of the last 131 amino acid of the protein and so comprises the last transmembrane domain and the C-terminal extremity. The expression of a truncated form of h N.I.S., lacking the last 131 amino acids, shows that this protein is not correctly addressed to the cell membrane and cells expressing this mutated symporter cannot accumulate iodide. However, our results show that the co-expression of the two N.I.S. parts, the truncated form lacking the last 131 amino acid, and the complementary C-terminal fragment, leads to cells presenting 10 % of the activity of cells expressing the whole N.I.S.. (author)

  12. Create a new vision for indigenous development

    Energy Technology Data Exchange (ETDEWEB)

    Chavez Alba, Rafael; Sanchez Arancibia, Oscar Armando [TRANSIERRA S.A., Santa Cruz (Bolivia)

    2009-07-01

    Transierra is a Bolivian company created in the year 2000 with the goal of transporting natural gas from the fields of San Alberto and San Antonio, in Tarija, to the Rio Grande Gas Compression Plant in Santa Cruz, for export to Brazil. Transierra has implemented a Social Action Plan, which allowed it to execute more than 800 community projects for the benefit of over 40 thousand families living in it's area of influence, with the presence of 146 indigenous communities, generally lagging behind in economic and productive life in the region and country. The Support Program to Guarani Development Plans (PA-PDG) is part of the Social Plan and is part of a long-term agreement signed between Transierra and indigenous organizations. The program has implemented more than one hundred projects for productive development, health, education, cultural revaluation, and strengthening organizational infrastructure, generating huge benefits in improving the living conditions of thousands of families of the Guarani people. This year a unique initiative was created with 4 Indigenous Captains and with the support of the International Finance Corporation (World Bank Group), including Business Plans to promote sustainable economic growth, created productive economic cycles involving improvements to the production and productivity to enter the commercial distribution of local and national markets. These four initiatives have meant a shift in the implementation and is helping to generate new dynamics in production, in addition to capturing significant resources from public and private investment, laying the groundwork for the improvement of the incomes and quality of life of its beneficiaries. (author)

  13. 10 ways to create shareholder value.

    Science.gov (United States)

    Rappaport, Alfred

    2006-09-01

    Executives have developed tunnel vision in their pursuit of shareholder value, focusing on short-term performance at the expense of investing in long-term growth. It's time to broaden that perspective and begin shaping business strategies in light of the competitive landscape, not the shareholder list. In this article, Alfred Rappaport offers ten basic principles to help executives create lasting shareholder value. For starters, companies should not manage earnings or provide earnings guidance; those that fail to embrace this first principle of shareholder value will almost certainly be unable to follow the rest. Additionally, leaders should make strategic decisions and acquisitions and carry assets that maximize expected value, even if near-term earnings are negatively affected as a result. During times when there are no credible value-creating opportunities to invest in the business, companies should avoid using excess cash to make investments that look good on the surface but might end up destroying value, such as ill-advised, overpriced acquisitions. It would be better to return the cash to shareholders in the form of dividends and buybacks. Rappaport also offers guidelines for establishing effective pay incentives at every level of management; emphasizes that senior executives need to lay their wealth on the line just as shareholders do; and urges companies to embrace full disclosure, an antidote to short-term earnings obsession that serves to lessen investor uncertainty, which could reduce the cost of capital and increase the share price. The author notes that a few types of companies--high-tech start-ups, for example, and severely capital-constrained organizations--cannot afford to ignore market pressures for short-term performance. Most companies with a sound, well-executed business model, however, could better realize their potential for creating shareholder value by adopting the ten principles.

  14. Creating a culture of mutual respect.

    Science.gov (United States)

    Kaplan, Kathryn; Mestel, Pamela; Feldman, David L

    2010-04-01

    The Joint Commission mandates that hospitals seeking accreditation have a process to define and address disruptive behavior. Leaders at Maimonides Medical Center, Brooklyn, New York, took the initiative to create a code of mutual respect that not only requires respectful behavior, but also encourages sensitivity and awareness to the causes of frustration that often lead to inappropriate behavior. Steps to implementing the code included selecting code advocates, setting up a system for mediating disputes, tracking and addressing operational system issues, providing training for personnel, developing a formal accountability process, and measuring the results. Copyright 2010 AORN, Inc. Published by Elsevier Inc. All rights reserved.

  15. Workshop: Creating Your Institutional Research Repository

    KAUST Repository

    Grenz, Daryl M.

    2016-11-08

    In 2002, the Scholarly Publishing and Academic Resources Coalition (SPARC) proposed the concept of an institutional repository to simultaneously disrupt and enhance the state of scholarly communications in the academic world. Thirteen years later, thousands of universities and other institutions have answered this call, but many more have not due to gaps in budgets, awareness and, most of all, practical guidance on creating an institutional repository. This workshop provides you with an essential primer on what it takes to establish a fully-functioning institutional repository. Every aspect of the process will be covered, including policies, procedures, staffing guidelines, workflows and repository technologies.

  16. Hospital culture--why create one?

    Science.gov (United States)

    Sovie, M D

    1993-01-01

    Hospitals, to survive, must be transformed into responsive, participative organizations capable of new practices that produce improved results in both quality of care and service at reduced costs. Creating, managing, and changing the culture are critical leadership functions that will enable the hospital to succeed. Strategic planning and effective implementation of planned change will produce the desired culture. Work restructuring, a focus on quality management along with changes in clinical practices, as well as the care and support processes, are all a part of the necessary hospital cultural revolution.

  17. Creating Posters for Effective Scientific Communication.

    Science.gov (United States)

    Bavdekar, Sandeep B; Vyas, Shruti; Anand, Varun

    2017-08-01

    A scientific poster is a summary of one's research that is presented in a visually engaging manner. Posters are presented as a means of short and quick scientific communications at conferences and scientific meetings. Presenting posters has advantages for the presenters and for conference attendees and organizers. It also plays a part in dissemination of research findings and furthering science. An effective poster is the one that focuses on a single message and conveys it through a concise and artistically attractive manner. This communication intends to provide tips on creating an effective poster to young scientists. © Journal of the Association of Physicians of India 2011.

  18. Do Treasure Islands Create Firm Value?

    DEFF Research Database (Denmark)

    Lai, Tat-kei; Ng, Travis

    They do! Otherwise, their use would not have been so prevalent among firms. How much firm value they create, however, is still an open question. Exploiting a political event in the U.K. that suddenly raised the cost of using tax havens, we find that there was a 0.87% reduction in cumulative...... abnormal return (CAR) among the sampled firms, corresponding to about £532 million in market capitalization. The firms of stronger corporate governance registered a stronger reduction in CAR. A simple linear extrapolation suggests that the firm value contributed by tax havens can be as much as £31 billion....

  19. ECONOMIC STRATEGY CREATED THROUGH THEORY FOCUSED PLANNING

    Directory of Open Access Journals (Sweden)

    Dumitru-Alexandru Bodislav

    2011-06-01

    Full Text Available The research done for this article is following the elaboration of a moldable model on the actual economic context, in which there are considered continuous learning elements on facts and commercial niches, evaluating a business starting from causality diagram and structuring its processes. Analyzing the evolution of a business and on the market in which they are deploying their activity through quantitative and qualitative measurement methods focused on trends. Testing hypotheses and business suppositions that are the main engine of theory focused on planning, a model of creating a strategic management process that will be developed in this article.

  20. Green electronics manufacturing creating environmental sensible products

    CERN Document Server

    Wang, John X

    2012-01-01

    Going ""green"" is becoming a major component of the mission for electronics manufacturers worldwide. While this goal seems simplistic, it poses daunting dilemmas. Yet, to compete effectively in the global economy, manufacturers must take the initiative to drive this crucial movement. Green Electronics Manufacturing: Creating Environmental Sensible Products provides you with a complete reference to design, develop, build, and install an electronic product with special consideration for the product's environmental impacts during its whole life cycle. The author discusses how to integrate the st

  1. Creating fluid injectivity in tar sands formations

    Science.gov (United States)

    Stegemeier, George Leo; Beer, Gary Lee; Zhang, Etuan

    2010-06-08

    Methods for treating a tar sands formation are described herein. Methods for treating a tar sands may include heating a portion of a hydrocarbon layer in the formation from one or more heaters located in the portion. The heat may be controlled to increase the permeability of at least part of the portion to create an injection zone in the portion with an average permeability sufficient to allow injection of a fluid through the injection zone. A drive fluid and/or an oxidizing fluid may be provided into the injection zone. At least some hydrocarbons are produced from the portion.

  2. Creating an anisotropic plasma resistivity with waves

    International Nuclear Information System (INIS)

    Fisch, N.J.; Boozer, A.H.

    1980-05-01

    An anisotropic plasma resistivity may be created by preferential heating of electrons traveling in one direction. This can result in a steady-state toroidal current in a tokamak even in the absence of net wave momentum. In fact, at high wave phase velocities, the current associated with the change in resistivity is greater than that associated with net momentum input. An immediate implication is that other waves, such as electron cyclotron waves, may be competitive with lower-hybrid waves as a means for generating current. An analytical expression is derived for the current generated per power dissipated which agrees remarkably well with numerical calculations

  3. Creating three-dimensional thermal maps

    CSIR Research Space (South Africa)

    Price

    2011-11-01

    Full Text Available stream_source_info Price_2011.pdf.txt stream_content_type text/plain stream_size 30895 Content-Encoding ISO-8859-1 stream_name Price_2011.pdf.txt Content-Type text/plain; charset=ISO-8859-1 Creating Three...-Dimensional Thermal Maps Mathew Price Cogency cc Cape Town Email: mathew@cogency.co.za Jeremy Green CSIR Centre for Mining Innovation Johannesburg Email: jgreen@csir.co.za John Dickens CSIR Centre for Mining Innovation Johannesburg Email: jdickens...

  4. ESTEEM manual. Deliverable 5 of Create Acceptance

    Energy Technology Data Exchange (ETDEWEB)

    Jolivet, E. [IAE, Toulouse (France); Mourik, R.; Raven, R.P.J.M.; Feenstra, C.F.J. [ECN Policy Studies, Petten (Netherlands); Alcantud Torrent, A.; Schaefer, B. [EcoInstitute, Barcelona (Spain); Heiskanen, E. [National Consumer Research Centre NCRC, Helsinki (Finland); Hodson, M. [Centre for Sustainable Urban and Regional Futures SURF, Manchester (United Kingdom); Oniszk-Poplawska, A. [Institute for Renewable Energy IEO, Warszawa (Poland); Difiore, M.; Fucsko, J. [Hungarian Environmental Economics Center MAKK, Budapest (Hungary); Maack, M.H. [Icelandic New Energy INE, Reykjavik (Iceland); Poti, B.M. [CERIS-CNR, Rome (Italy); Prasad, G. [University of Cape Town UCT, Capetown (South Africa); Brohmann, B.; Fritsche, U.R.; Huenecke, K. [OEKO Institut, Freiburg (Germany)

    2008-05-15

    The ESTEEM tool is one of the outcomes of Create Acceptance. ESTEEM (Engage stake-holders through a systematic toolbox to manage new energy projects) is a six step tool which is performed by a consultant in close cooperation with the project manager of a new energy project. The focus of the tool is put on the early recognition and discussion of stakeholders expectations and the integration of these in the design of the project. ESTEEM, including background information is freely available via www.esteem-tool.eu.

  5. Nuclear power creates new possibilities for marketing

    International Nuclear Information System (INIS)

    Van Vuren, K.

    1978-01-01

    Irradiation of food presents various advantages over other forms of food sterilization and preservation: 1) food is disinfested of insects, microorganisms and parasites without harming the fooddstuff itself; 2) food is handled in its packaged form when irradiated; 3) the irradiation process is a 'cold' process, i.e. foodstuff need not undergo a rise in temperature; this prevents harming of the tissues; and 4) the storage life is increased, creating new possibilities for marketing and trade. A few experiments with vegetables, fruit and meat are described

  6. Creating and probing coherent atomic states

    Energy Technology Data Exchange (ETDEWEB)

    Reinhold, C.O.; Burgdoerfer, J. [Oak Ridge National Lab., TN (United States). Physics Div.]|[Univ. of Tennessee, Knoxville, TN (United States). Dept. of Physics and Astronomy; Frey, M.T.; Dunning, F.B. [Rice Univ., Houston, TX (United States)

    1997-06-01

    The authors present a brief review of recent experimental and theoretical time resolved studies of the evolution of atomic wavepackets. In particular, wavepackets comprising a superposition of very-high-lying Rydberg states which are created either using a short half-cycle pulse (HCP) or by rapid application of a DC field. The properties of the wavepackets are probed using a second HCP that is applied following a variable time delay and ionizes a fraction of the atoms, much like a passing-by ion in atomic collisions.

  7. Isolation and expression of a novel chick G-protein cDNA coding for a G alpha i3 protein with a G alpha 0 N-terminus.

    OpenAIRE

    Kilbourne, E J; Galper, J B

    1994-01-01

    We have cloned cDNAs coding for G-protein alpha subunits from a chick brain cDNA library. Based on sequence similarity to G-protein alpha subunits from other eukaryotes, one clone was designated G alpha i3. A second clone, G alpha i3-o, was identical to the G alpha i3 clone over 932 bases on the 3' end. The 5' end of G alpha i3-o, however, contained an alternative sequence in which the first 45 amino acids coded for are 100% identical to the conserved N-terminus of G alpha o from species such...

  8. Creating a culture where employee engagement Thrives

    Energy Technology Data Exchange (ETDEWEB)

    Don Groover, C.S.P. [Behavioral Science Technology, Ojai, CA (United States)

    2007-07-01

    Safety leaders across industries face a critical challenge: engaging employees. While engagement of a few people may be easy in short-term projects, it is significantly more difficult with long-term processes. In this session we show leaders how they can create a culture where workers are more open and even eager to be involved in safety efforts. Our experience with safety leaders in the nuclear industry has verified that when the factors that drive organizational functioning are understood, leaders are enabled to augment employee engagement and attain significant improvement in safety outcomes. The underlying factors that influence employee engagement, performance, outcomes, and organizational culture are the same the world over. We will also show how safety is capable, by its intrinsic value, of winning profound support and direct engagement of employees. In this session, we will examine how leaders can leverage their decisions and actions to win over employees to safety and support them in their endeavors to promote it. Using the safety leadership best practices Vision, Credibility, Accountability, Communication, Collaboration, Action Orientation, and Recognition and Feedback, leaders increase their impact on their organization in favor of a culture that supports safety and employee engagement. Leaders that create a climate and culture where employee engagement thrives, realize better safety results. Leadership is not exclusively an inborn talent; it can be developed and enhanced. To this end, we will also show the advantages of transformational leadership style by comparing it to more classical transactional leadership.

  9. Data Programming: Creating Large Training Sets, Quickly

    Science.gov (United States)

    Ratner, Alexander; De Sa, Christopher; Wu, Sen; Selsam, Daniel; Ré, Christopher

    2018-01-01

    Large labeled training sets are the critical building blocks of supervised learning methods and are key enablers of deep learning techniques. For some applications, creating labeled training sets is the most time-consuming and expensive part of applying machine learning. We therefore propose a paradigm for the programmatic creation of training sets called data programming in which users express weak supervision strategies or domain heuristics as labeling functions, which are programs that label subsets of the data, but that are noisy and may conflict. We show that by explicitly representing this training set labeling process as a generative model, we can “denoise” the generated training set, and establish theoretically that we can recover the parameters of these generative models in a handful of settings. We then show how to modify a discriminative loss function to make it noise-aware, and demonstrate our method over a range of discriminative models including logistic regression and LSTMs. Experimentally, on the 2014 TAC-KBP Slot Filling challenge, we show that data programming would have led to a new winning score, and also show that applying data programming to an LSTM model leads to a TAC-KBP score almost 6 F1 points over a state-of-the-art LSTM baseline (and into second place in the competition). Additionally, in initial user studies we observed that data programming may be an easier way for non-experts to create machine learning models when training data is limited or unavailable. PMID:29872252

  10. Creating a culture where employee engagement Thrives

    International Nuclear Information System (INIS)

    Don Groover, C.S.P.

    2007-01-01

    Safety leaders across industries face a critical challenge: engaging employees. While engagement of a few people may be easy in short-term projects, it is significantly more difficult with long-term processes. In this session we show leaders how they can create a culture where workers are more open and even eager to be involved in safety efforts. Our experience with safety leaders in the nuclear industry has verified that when the factors that drive organizational functioning are understood, leaders are enabled to augment employee engagement and attain significant improvement in safety outcomes. The underlying factors that influence employee engagement, performance, outcomes, and organizational culture are the same the world over. We will also show how safety is capable, by its intrinsic value, of winning profound support and direct engagement of employees. In this session, we will examine how leaders can leverage their decisions and actions to win over employees to safety and support them in their endeavors to promote it. Using the safety leadership best practices Vision, Credibility, Accountability, Communication, Collaboration, Action Orientation, and Recognition and Feedback, leaders increase their impact on their organization in favor of a culture that supports safety and employee engagement. Leaders that create a climate and culture where employee engagement thrives, realize better safety results. Leadership is not exclusively an inborn talent; it can be developed and enhanced. To this end, we will also show the advantages of transformational leadership style by comparing it to more classical transactional leadership

  11. Creating a Regional Healthcare Network: People First.

    Science.gov (United States)

    Michel-Verkerke, Margreet B

    2016-01-01

    Care organizations in the Dutch region Apeldoorn want to collaborate more in order to improve the care provision to elderly and psychiatric patients living independently. In order to support the collaboration they intend to create a regional digital healthcare network. The research was focused on the relevance of a regional healthcare network for care providers. Eleven semi-structured interviews based on the USE IT-model, were conducted with care providers and staff members. Results show that care providers need to tune their activities for this target group and create an agreement on integrated care. The relevance of a digital communication and collaboration platform is high. The regional healthcare network should support the collaboration between care providers by: 1. Offering a communication platform to replace the time consuming communication by telephone; 2. Making patient information available for patient and care provider at patients' homes; 3. Giving insight in who is giving what care to whom; and 4. Giving access to knowledge about the target group: elderly and psychiatric patients living independently.

  12. Cell adhesion pattern created by OSTE polymers.

    Science.gov (United States)

    Liu, Wenjia; Li, Yiyang; Ding, Xianting

    2017-04-24

    Engineering surfaces with functional polymers is a crucial issue in the field of micro/nanofabrication and cell-material interface studies. For many applications of surface patterning, it does not need cells to attach on the whole surface. Herein, we introduce a novel polymer fabrication protocol of off-stoichiometry thiol-ene (OSTE) polymers to create heterogeneity on the surface by utilizing 3D printing and soft-lithography. By choosing two OSTE polymers with different functional groups, we create a pattern where only parts of the surface can facilitate cell adhesion. We also study the hydrophilic property of OSTE polymers by mixing poly(ethylene glycol) (PEG) directly with pre-polymers and plasma treatments afterwards. Moreover, we investigate the effect of functional groups' excess ratio and hydrophilic property on the cell adhesion ability of OSTE polymers. The results show that the cell adhesion ability of OSTE materials can be tuned within a wide range by the coupling effect of functional groups' excess ratio and hydrophilic property. Meanwhile, by mixing PEG with pre-polymers and undergoing oxygen plasma treatment afterward can significantly improve the hydrophilic property of OSTE polymers.

  13. Our Lunar Destiny: Creating a Lunar Economy

    Science.gov (United States)

    Rohwer, Christopher J.

    2000-01-01

    "Our Lunar Destiny: Creating a Lunar Economy" supports a vision of people moving freely and economically between the earth and the Moon in an expansive space and lunar economy. It makes the economic case for the creation of a lunar space economy and projects the business plan that will make the venture an economic success. In addition, this paper argues that this vision can be created and sustained only by private enterprise and the legal right of private property in space and on the Moon. Finally, this paper advocates the use of lunar land grants as the key to unleashing the needed capital and the economic power of private enterprise in the creation of a 21st century lunar space economy. It is clear that the history of our United States economic system proves the value of private property rights in the creation of any new economy. It also teaches us that the successful development of new frontiers-those that provide economic opportunity for freedom-loving people-are frontiers that encourage, respect and protect the possession of private property and the fruits of labor and industry. Any new 21st century space and lunar economy should therefore be founded on this same principle.

  14. Creating Sister Cities: An Exchange Across Hemispheres

    Science.gov (United States)

    Adams, M. T.; Cabezon, S. A.; Hardy, E.; Harrison, R. J.

    2008-06-01

    Sponsored by Associated Universities, Inc. (AUI) and the National Radio Astronomy Observatory (NRAO), this project creates a cultural and educational exchange program between communities in South and North America, linking San Pedro de Atacama in Chile and Magdalena, New Mexico in the United States. Both communities have similar demographics, are in relatively undeveloped regions of high-elevation desert, and are located near major international radio astronomy research facilities. The Atacama Large Millimeter/submillimeter Array (ALMA) is just 40 km east of San Pedro; the Very Large Array (VLA) is just 40 km west of Magdalena. In February 2007, the Mayor of San Pedro and two teachers visited Magdalena for two weeks; in July 2007 three teachers from Magdalena will visit San Pedro. These visits enable the communities to lay the foundation for a permanent, unique partnership. The teachers are sharing expertise and teaching methodologies for physics and astronomy. In addition to creating science education opportunities, this project offers students linguistic and cultural connections. The town of San Pedro, Chile, hosts nearly 100,000 tourists per year, and English language skills are highly valued by local students. Through exchanges enabled by email and distance conferencing, San Pedro and Magdalena students will improve English and Spanish language skills while teaching each other about science and their respective cultures. This poster describes the AUI/NRAO Sister Cities program, including the challenges of cross-cultural communication and the rewards of interpersonal exchanges between continents and cultures.

  15. Creating a Multi-axis Machining Postprocessor

    Directory of Open Access Journals (Sweden)

    Petr Vavruška

    2012-01-01

    Full Text Available This paper focuses on the postprocessor creation process. When using standard commercially available postprocessors it is often very difficult to modify its internal source code, and it is a very complex process, in many cases even impossible, to implement the newly-developed functions. It is therefore very important to have a method for creating a postprocessor for any CAM system, which allows CL data (Cutter Location data to be generated to a separate text file. The goal of our work is to verify the proposed method for creating a postprocessor. Postprocessor functions for multi-axis machiningare dealt with in this work. A file with CL data must be translated by the postprocessor into an NC program that has been customized for a specific production machine and its control system. The postprocessor is therefore verified by applications for machining free-form surfaces of complex parts, and by executing the NC programs that are generated on real machine tools. This is also presented here.

  16. Creating a sustainable energy future for Australia

    International Nuclear Information System (INIS)

    Sonneborn, C.L.

    1995-01-01

    A joint industry approach is needed to put in place a sustainable energy system that is economically and technologically feasible. The industry sectors involved must include the renewable energy industry, energy efficiency industry and the natural gas industry. Conventional forecasts of energy futures make far less use of these industries than is economically and technically feasible. Existing forecasts make the trade off between acceptable levels of economic growth, limitation of greenhouse gases and dependence on coal and oil appear more difficult than they actually are and overlook the benefits of sustainable energy industry development. This paper outlines how national gains from carefully targeted action can exceed national losses while substantially reducing greenhouse gases and creating jobs at zero or negative costs. (author). 3 figs., 27 refs

  17. Creating innovative programs for the future.

    Science.gov (United States)

    Allen, Patricia E; Keough, Vicki A; Armstrong, Myrna L

    2013-09-01

    Although several major national mandates advocate for a better educated workforce, this push comes at a time when the competition for faculty, financial resources, advanced technology, and students remains strong. If nurse educators are seriously considering creating a new nurse program at their school, some key points are essential during the development stage. Using the innovation frameworks from the Institute of Healthcare Improvement, from the global design firm IDEO, and from Gladwell's The Tipping Point: How Little Things Can Make a Big Difference, this article examines the informal, formal, internal, and external work needed during program conceptualization, initial program exploration, resource infrastructure, support, and evaluation for an effective and innovative plan. Copyright 2013, SLACK Incorporated.

  18. Creating biological nanomaterials using synthetic biology

    International Nuclear Information System (INIS)

    Rice, MaryJoe K; Ruder, Warren C

    2014-01-01

    Synthetic biology is a new discipline that combines science and engineering approaches to precisely control biological networks. These signaling networks are especially important in fields such as biomedicine and biochemical engineering. Additionally, biological networks can also be critical to the production of naturally occurring biological nanomaterials, and as a result, synthetic biology holds tremendous potential in creating new materials. This review introduces the field of synthetic biology, discusses how biological systems naturally produce materials, and then presents examples and strategies for incorporating synthetic biology approaches in the development of new materials. In particular, strategies for using synthetic biology to produce both organic and inorganic nanomaterials are discussed. Ultimately, synthetic biology holds the potential to dramatically impact biological materials science with significant potential applications in medical systems. (review)

  19. Creating a false memory in the hippocampus.

    Science.gov (United States)

    Ramirez, Steve; Liu, Xu; Lin, Pei-Ann; Suh, Junghyup; Pignatelli, Michele; Redondo, Roger L; Ryan, Tomás J; Tonegawa, Susumu

    2013-07-26

    Memories can be unreliable. We created a false memory in mice by optogenetically manipulating memory engram-bearing cells in the hippocampus. Dentate gyrus (DG) or CA1 neurons activated by exposure to a particular context were labeled with channelrhodopsin-2. These neurons were later optically reactivated during fear conditioning in a different context. The DG experimental group showed increased freezing in the original context, in which a foot shock was never delivered. The recall of this false memory was context-specific, activated similar downstream regions engaged during natural fear memory recall, and was also capable of driving an active fear response. Our data demonstrate that it is possible to generate an internally represented and behaviorally expressed fear memory via artificial means.

  20. Creating Effective Dialogue Around Climate Change

    Science.gov (United States)

    Kiehl, J. T.

    2015-12-01

    Communicating climate change to people from diverse sectors of society has proven to be difficult in the United States. It is widely recognized that difficulties arise from a number of sources, including: basic science understanding, the psychologically affect laden content surrounding climate change, and the diversity of value systems that exist in our society. I explore ways of working with the affect that arises around climate change and describe specific methods to work with the resistance often encountered when communicating this important issue. The techniques I describe are rooted in psychology and group process and provide means for creating more effective narratives to break through the barriers to communicating climate change science. Examples are given from personal experiences in presenting climate change to diverse groups.

  1. Minkowski metrics in creating universal ranking algorithms

    Directory of Open Access Journals (Sweden)

    Andrzej Ameljańczyk

    2014-06-01

    Full Text Available The paper presents a general procedure for creating the rankings of a set of objects, while the relation of preference based on any ranking function. The analysis was possible to use the ranking functions began by showing the fundamental drawbacks of commonly used functions in the form of a weighted sum. As a special case of the ranking procedure in the space of a relation, the procedure based on the notion of an ideal element and generalized Minkowski distance from the element was proposed. This procedure, presented as universal ranking algorithm, eliminates most of the disadvantages of ranking functions in the form of a weighted sum.[b]Keywords[/b]: ranking functions, preference relation, ranking clusters, categories, ideal point, universal ranking algorithm

  2. Creating Inclusive Youth Programs for LGBTQ+ Communities

    Directory of Open Access Journals (Sweden)

    Katherine E. Soule

    2017-06-01

    Full Text Available It is vital for youth to experience inclusive programming that is welcoming. Extension has a responsibility and an obligation to provide youth with programs and spaces that are inclusive of all sexes, gender identities, gender expressions, and sexual orientations. This article provides an overview of appropriate terminology, as well as steps for creating inclusive Extension spaces and programs for youth who identify as members of lesbian, gay, bisexual, transgender, or queer/questioning (LGBTQ+ communities. With a focus on urban Extension audiences, this article uses accessible language, self-reflective prompts, and supporting visual aids to share lessons learned from ongoing inclusivity trainings with Extension personnel across the nation, as well as from research activities and inclusive programming.

  3. Challenging Unjust Authority and Creating a Profession.

    Science.gov (United States)

    Doona, Mary Ellen

    2017-05-01

    The emergence of the trained nurse as a new profession for women coincided with the transformation of hospitals from refuges for the sick and poor into treatment facilities. The hospital was an apt setting for learning nursing's fundamentals, but the domestic demands of the setting soon took precedence over nursing education. Nursing superintendents struggled to honor the right of pupil nurses to an education over the might of the hospital's demands. The 20th anniversary of the Trained Nurse Movement in 1893 proved the decisive moment when nursing superintendents created professional associations and gathered the nursing diaspora into a coherent force. Recourse to the law followed with nurses seeking legal validation of their authority over nursing education. Now, as then, a nurse's authority is a right that has to be protected when the might of individuals and institutions, both subtle and blatant, interferes with nurses caring for the well-being of society.

  4. Creating Visual Design and Meaningful Audience Experiences

    DEFF Research Database (Denmark)

    Steijn, Arthur; Ion Wille, Jakob

    2014-01-01

    The main purpose of the EU Interreg funded Classical Composition Music and Experience Design project, was to rethink audience experiences and develop knowledge of applied technologies connected to classical music and live concerts. The project and its main objectives was motivated by at least thee...... conditions. The most important being 1) the development in new technology creating new expectations in audiences attending cultural events, including classical concerts, 2) resent decline in audiences attending classical music and 3) a will to strengthen relations between cultural institutions, creative...... businesses and educational institutions in the Øresund region (including the city and surroundings of Malmø and Copenhagen). Therefore the project Classical Composition Music and Experience Design focused on developing new and meaningful audience experiences where live classical music meets new digital...

  5. Creating marketing strategies for higher education institutions

    Directory of Open Access Journals (Sweden)

    Lidia Białoń

    2015-12-01

    Full Text Available The article presents a thesis that the primary premise of creating marketing strategies for higher education institution is a three-dimensional notion of marketing. The first dimension lies in the theoretical notions of the essence of marketing, including the transactional marketing (1.0, relationship marketing (2.0 and spiritual marketing (3.0. The second dimension is formed by methods of marketing research and accurate notions of marketing, while the third are channels of marketing information. Harmonizing these three dimensions is a precondition for effective marketing. Among other conditions for effective strategies there are: aligning goals of the chosen strategy with the mission of higher education institution, correct choice of targeted segments of the market and of marketing tools. The article also gives a sample classification of marketing strategies based on these criteria, with emphasis on the fact, that every higher education institution employs its own strategy.

  6. How to create high-performing teams.

    Science.gov (United States)

    Lam, Samuel M

    2010-02-01

    This article is intended to discuss inspirational aspects on how to lead a high-performance team. Cogent topics discussed include how to hire staff through methods of "topgrading" with reference to Geoff Smart and "getting the right people on the bus" referencing Jim Collins' work. In addition, once the staff is hired, this article covers how to separate the "eagles from the ducks" and how to inspire one's staff by creating the right culture with suggestions for further reading by Don Miguel Ruiz (The four agreements) and John Maxwell (21 Irrefutable laws of leadership). In addition, Simon Sinek's concept of "Start with Why" is elaborated to help a leader know what the core element should be with any superior culture. Thieme Medical Publishers.

  7. Creating biological nanomaterials using synthetic biology.

    Science.gov (United States)

    Rice, MaryJoe K; Ruder, Warren C

    2014-02-01

    Synthetic biology is a new discipline that combines science and engineering approaches to precisely control biological networks. These signaling networks are especially important in fields such as biomedicine and biochemical engineering. Additionally, biological networks can also be critical to the production of naturally occurring biological nanomaterials, and as a result, synthetic biology holds tremendous potential in creating new materials. This review introduces the field of synthetic biology, discusses how biological systems naturally produce materials, and then presents examples and strategies for incorporating synthetic biology approaches in the development of new materials. In particular, strategies for using synthetic biology to produce both organic and inorganic nanomaterials are discussed. Ultimately, synthetic biology holds the potential to dramatically impact biological materials science with significant potential applications in medical systems.

  8. Creating an interactive environment for pediatric assessment.

    Science.gov (United States)

    de Armas Weber, D; Easley-Rosenberg, A

    2001-01-01

    An interactive assessment room (IAR) was conceived to explore the effect of a dynamic environment on the pediatric assessment process and subsequent individualized goal development. Selection of a central theme, creation of a multipurpose space, provision of multisensory experiences, maximization of environmental affordances, provision of effective motivators and opportunities for goal attainment, and facilitation of a transdisciplinary assessment were identified as integral to designing the IAR. A central farm theme was selected to create five task-oriented activity stations. The IAR offered a creative assessment environment for transdisciplinary, practice-based application of current motor development and behavioral models. In addition, the IAR facilitated exploratory play essential to promoting the client's optimal performance to arrive at the development of appropriate treatment goals.

  9. Consistency between recognition and behavior creates consciousness

    Directory of Open Access Journals (Sweden)

    Keita Inaba

    2004-08-01

    Full Text Available What is consciousness? Is it possible to create consciousness mechanically? Various studies have been performed in the fields of psychology and cerebral science to answer these questions. As of yet, however, no researchers have proposed a model capable of explaining the mind-body problem described by Descartes or replicating a consciousness as advanced as that of human beings. Ancient people believed that the consciousness resided in a Homunculus, a human in miniature who lived in the brain. It is no mystery that the ancients came up with such an idea; for consciousness has always been veiled in mystery, beyond the reach of our explorative powers. We can assert, however, that consciousness does not "live" in us, but "exists" in us. Insofar as the processes occurring inside the human brain are a product of the physical activity of the neurons that reside there, we believe that it should be possible to define consciousness systematically.

  10. Tissue loading created during spinal manipulation in comparison to loading created by passive spinal movements.

    Science.gov (United States)

    Funabashi, Martha; Kawchuk, Gregory N; Vette, Albert H; Goldsmith, Peter; Prasad, Narasimha

    2016-12-01

    Spinal manipulative therapy (SMT) creates health benefits for some while for others, no benefit or even adverse events. Understanding these differential responses is important to optimize patient care and safety. Toward this, characterizing how loads created by SMT relate to those created by typical motions is fundamental. Using robotic testing, it is now possible to make these comparisons to determine if SMT generates unique loading scenarios. In 12 porcine cadavers, SMT and passive motions were applied to the L3/L4 segment and the resulting kinematics tracked. The L3/L4 segment was removed, mounted in a parallel robot and kinematics of SMT and passive movements replayed robotically. The resulting forces experienced by L3/L4 were collected. Overall, SMT created both significantly greater and smaller loads compared to passive motions, with SMT generating greater anterioposterior peak force (the direction of force application) compared to all passive motions. In some comparisons, SMT did not create significantly different loads in the intact specimen, but did so in specific spinal tissues. Despite methodological differences between studies, SMT forces and loading rates fell below published injury values. Future studies are warranted to understand if loading scenarios unique to SMT confer its differential therapeutic effects.

  11. Winter barley mutants created in the Ukraine

    International Nuclear Information System (INIS)

    Zayats, O.M.

    2001-01-01

    Full text: Increasing fodder and protein production is one of the objectives of the development of agriculture in Ukraine. Higher productivity of fodder crops, due to new highly productive varieties, is the means to meet this aim. Winter barley is an important crop for fodder purposes. The climate of the Ukraine is favourable for growing this crop. The areas used for the growth of winter barley are however, small (500,000-550,000 ha) and there is a shortage of good quality varieties. The main aim of the work was therefore to create new varieties of highly productive winter barley, of good quality. The new varieties and mutation lines of winter barley were created under the influence of water solutions of N-nitroso-N-methylurea (NMH - 0,012, 0,005%), N-nitroso-N-ethylurea (NEH - 0,05; 0.025; 0,012%) ethyleneimine (EI - 0,02; 0,01; 0,005%) on winter barley seeds of the varieties of local and foreign selections. On the basis of many years of investigations (1984-94) the following mutations were described: hard-grained, winter-hardiness, earliness, middle-maturity, late-maturity, wide and large leaves, narrow leaves, multinodal, great number of leaves, great number of flowers, strong stem (lodging resistant), tallness, semi-dwarfness, dwarfness, and high productivity. Particularly valuable are mutants with high productivity of green bulk. Their potential yield is 70 t/ha. As a result of the work two varieties of winter barley 'Shyrokolysty' and 'Kormovy' were released into the State register of plant varieties of the Ukraine. The other valuable mutant genotypes are used in cross breeding programmes. (author)

  12. Phactr3/scapinin, a member of protein phosphatase 1 and actin regulator (phactr family, interacts with the plasma membrane via basic and hydrophobic residues in the N-terminus.

    Directory of Open Access Journals (Sweden)

    Akihiro Itoh

    Full Text Available Proteins that belong to the protein phosphatase 1 and actin regulator (phactr family are involved in cell motility and morphogenesis. However, the mechanisms that regulate the actin cytoskeleton are poorly understood. We have previously shown that phactr3, also known as scapinin, localizes to the plasma membrane, including lamellipodia and membrane ruffles. In the present study, experiments using deletion and point mutants showed that the basic and hydrophobic residues in the N-terminus play crucial roles in the localization to the plasma membrane. A BH analysis (http://helixweb.nih.gov/bhsearch is a program developed to identify membrane-binding domains that comprise basic and hydrophobic residues in membrane proteins. We applied this program to phactr3. The results of the BH plot analysis agreed with the experimentally determined region that is responsible for the localization of phactr3 to the plasma membrane. In vitro experiments showed that the N-terminal itself binds to liposomes and acidic phospholipids. In addition, we showed that the interaction with the plasma membrane via the N-terminal membrane-binding sequence is required for phactr3-induced morphological changes in Cos7 cells. The membrane-binding sequence in the N-terminus is highly conserved in all members of the phactr family. Our findings may provide a molecular basis for understanding the mechanisms that allow phactr proteins to regulate cell morphogenesis.

  13. Dancing Lights: Creating the Aurora Story

    Science.gov (United States)

    Wood, E. L.; Cobabe-Ammann, E. A.

    2009-12-01

    Science tells a story about our world, our existence, our history, and the larger environment our planet occupies. Bearing this in mind, we created a series of lessons for 3rd-5th grades using a cross-disciplinary approach to teaching about the aurora by incorporating stories, photos, movies, and geography into the science in order to paint a broad picture and answer the question, “why do we care?” The fundamental backbone of the program is literacy. Students write and illustrate fiction and non-fiction work, poetry, and brochures that solidify both language arts skills and science content. In a time when elementary teachers relegate science to less than one hour per week, we have developed a novel science program that can be easily integrated with other topics during the typical school day to increase the amount of science taught in a school year. We are inspiring students to take an interest in the natural world with this program, a stepping-stone for larger things.

  14. When and how to create strategic alliances

    International Nuclear Information System (INIS)

    Reinsvold, C.

    1994-01-01

    As oil prices remain flat and discoveries become more costly, the relationships between suppliers and operators are changing. Arms-length, unit pricing buying practices remain the norm for many buyers, but others have identified advantages of forming alliances that make greater use of the suppliers' capabilities. Recently, operators and the service industry have started to evolve a new culture that provides value-added solutions in an incentive environment. coupled with long-term risk-sharing or partnering arrangements, the potential advantages to both the operator and supplier are clear. An alliance is a long-term commitment between the operator and one or more supplier organizations, formed to achieve well-defined project objectives in a way that is beneficial to all alliance partners. Alliances take many forms and are adapted to meet the needs of the project. Successful alliances are characterized by trust, common goals and open communication. They build on the efficiencies, capabilities and strengths each partner brings to the relationship. The nine steps to creating a successful alliance are described

  15. Creating energy citizenship through material participation.

    Science.gov (United States)

    Ryghaug, Marianne; Skjølsvold, Tomas Moe; Heidenreich, Sara

    2018-04-01

    Transitions towards low-carbon energy systems will be comprehensive and demanding, requiring substantial public support. One important contribution from STS is to highlight the roles of citizens and public engagement. Until recently, energy users have often been treated as customers and passive market actors, or as recipients of technology at the margins of centralized systems. With respect to the latter role, critical or hesitant public action has been explained in terms of NIMBYism and knowledge deficits. This article focuses on the production of energy citizenship when considering public participation in low-carbon energy transitions. We draw upon the theory of 'material participation' to highlight how introducing and using emergent energy technologies may create new energy practices. We analyze an ongoing introduction of new material objects, highlighting the way these technologies can be seen as material interventions co-constructing temporalities of new and sustainable practices. We argue that artefacts such as the electric car, the smart meter and photovoltaic panels may become objects of participation and engagement, and that the introduction of such technologies may foster material participation and energy citizenship. The paper concludes with a discussion about the role of policies for low-carbon energy transitions on the making of energy citizenship, as well as limits of introducing a materially based energy citizenship.

  16. Creating a Masters in Numeracy Program

    Directory of Open Access Journals (Sweden)

    Eric Gaze

    2010-07-01

    Full Text Available The Master of Science in Numeracy program at Alfred University received full approval from the New York State Education Department (NYSED in May of 2007. This first-of-its-kind program seeks to provide teachers at all levels, from across the curriculum, the skills, and more importantly the confidence, to introduce relevant quantitative concepts in their own disciplines. Created to be a complement of the MS Ed. in Literacy, the 30-hour MS in Numeracy program consists of four required core courses (Teaching Numeracy, Teaching with Data, Assessment and Learning Theories in Numeracy, and Doing Science and Numeracy, five electives from a list of numeracy and literacy courses, and a Masters project. The program graduated its first student in May 2008 and three more since then. Major challenges for the program have included the uncertain (i.e., by-application connection between an MS and licensure (in contrast to the automatic professional certification for MS Ed. degrees and the small number of faculty involved in teaching the numeracy courses. The current status of the program is questionable as the person (the author who taught the first three core courses has left the University and has not yet been replaced. Even so, I believe this MS in Numeracy program offers a potentially useful example of a strategy to enhance the spread of QL through teacher preparation.

  17. Creating the optimized international marketing mix

    Directory of Open Access Journals (Sweden)

    Grubor Aleksandar

    2005-01-01

    Full Text Available In the international practice of Serbian companies still dominant approach is based on traditional foreign trade approach, which in the significant dimension ballast effective inclusion of domestic companies in the contemporary business. Such practice is a result of the modality of international trading evolved and stimulated recently including long lasting period of self managing socialism-business as well international orientation in trading was originated by the state support, which procured in the edification of so-called national corporations as a chief exporter. Long-time existing of that modality of the international trading procured in the shrinking and limitation possibilities for comprehensive development of the international orientation of domestic i.e local companies, which is an argument of objectively difficult international position of Serbian firms in the moment. Paper discuss different issues including assessment that incoming presence of regional or international investors on the domestic market will proffer possibilities for enhancement a domestic management and marketing know - how among others. So, this is a way how local management circles could make substantial progress in creating the optimized international marketing mix within domestic companies following on-going business experience.

  18. Creating cultures of excellence: Strategies and outcomes

    Directory of Open Access Journals (Sweden)

    Michael Mintrom

    2014-12-01

    Full Text Available Research findings on effective support for learning, the development of expertise, and the psychology of success suggest that the pursuit of excellence is teachable. Within the emerging field of research and practice termed “the scholarship of teaching and learning,” considerable effort has been made to document the practices of teachers who, by various measures, have been deemed excellent. In contrast, no effort has been made to codify how students can be trained to self-consciously build behaviors that generate excellent outcomes. This article reports on a multi-year effort to create cultures of excellence among cohorts of graduate students. A statistical analysis of subsequent student performance on a significant, related task indicates that explicitly promoting a culture of excellence among course participants can have a positive and sustained impact on their individual practices. Comments from subsequent student reflections further support this claim. The teaching strategies reported here could be refined, replicated, and reinvented to good effect across higher education. They are also of special relevance to those delivering professional development training to early- and mid-career professionals.

  19. Creating a Universe, a Conceptual Model

    Directory of Open Access Journals (Sweden)

    James R. Johnson

    2016-10-01

    Full Text Available Space is something. Space inherently contains laws of nature: universal rules (mathematics, space dimensions, types of forces, types of fields, and particle species, laws (relativity, quantum mechanics, thermodynamics, and electromagnetism and symmetries (Lorentz, Gauge, and symmetry breaking. We have significant knowledge about these laws of nature because all our scientific theories assume their presence. Their existence is critical for developing either a unique theory of our universe or more speculative multiverse theories. Scientists generally ignore the laws of nature because they “are what they are” and because visualizing different laws of nature challenges the imagination. This article defines a conceptual model separating space (laws of nature from the universe’s energy source (initial conditions and expansion (big bang. By considering the ramifications of changing the laws of nature, initial condition parameters, and two variables in the big bang theory, the model demonstrates that traditional fine tuning is not the whole story when creating a universe. Supporting the model, space and “nothing” are related to the laws of nature, mathematics and multiverse possibilities. Speculation on the beginning of time completes the model.

  20. Dental practice websites: creating a Web presence.

    Science.gov (United States)

    Miller, Syrene A; Forrest, Jane L

    2002-07-01

    Web technology provides an opportunity for dentists to showcase their practice philosophy, quality of care, office setting, and staff in a creative manner. Having a Website provides a practice with innovative and cost-effective communications and marketing tools for current and potential patients who use the Internet. The main benefits of using a Website to promote one's practice are: Making office time more productive, tasks more timely, follow-up less necessary Engaging patients in an interactive and visual learning process Providing online forms and procedure examples for patients Projecting a competent and current image Tracking the usage of Web pages. Several options are available when considering the development of a Website. These options range in cost based on customization of the site and ongoing support services, such as site updates, technical assistance, and Web usage statistics. In most cases, Websites are less expensive than advertising in the phone book. Options in creating a Website include building one's own, employing a company that offers Website templates, and employing a company that offers customized sites. These development options and benefits will continue to grow as individuals access the Web and more information and sites become available.

  1. Creating conditions for cooperative learning: Basic elements

    Directory of Open Access Journals (Sweden)

    Ševkušić-Mandić Slavica G.

    2003-01-01

    Full Text Available Although a large number of research evidence speak out in favor of cooperative learning, its effectiveness in teaching does not depend only on teacher’s and students’ enthusiasm and willingness to work in such a manner. Creating cooperative situations in learning demands a serious preparation and engagement on the part of teacher who is structuring various aspects of work in the classroom. Although there exist a large number of models and techniques of cooperative learning, which vary in the way in which students work together, in the structure of learning tasks as well as in the degree to which cooperative efforts of students are coupled with competition among groups, some elements should be present in the structure of conditions irrespective of the type of group work in question. Potential effects of cooperation are not likely to emerge unless teachers apply five basic elements of cooperative structure: 1. structuring of the learning task and students’ positive interdependence, 2. individual responsibility, 3. upgrading of "face to face" interaction, 4. training of students’ social skills, and 5. evaluation of group processes. The paper discusses various strategies for establishing the mentioned elements and concrete examples for teaching practice are provided, which should be of assistance to teachers for as much successful cooperative learning application as possible in work with children.

  2. Leveraging Metadata to Create Interactive Images... Today!

    Science.gov (United States)

    Hurt, Robert L.; Squires, G. K.; Llamas, J.; Rosenthal, C.; Brinkworth, C.; Fay, J.

    2011-01-01

    The image gallery for NASA's Spitzer Space Telescope has been newly rebuilt to fully support the Astronomy Visualization Metadata (AVM) standard to create a new user experience both on the website and in other applications. We encapsulate all the key descriptive information for a public image, including color representations and astronomical and sky coordinates and make it accessible in a user-friendly form on the website, but also embed the same metadata within the image files themselves. Thus, images downloaded from the site will carry with them all their descriptive information. Real-world benefits include display of general metadata when such images are imported into image editing software (e.g. Photoshop) or image catalog software (e.g. iPhoto). More advanced support in Microsoft's WorldWide Telescope can open a tagged image after it has been downloaded and display it in its correct sky position, allowing comparison with observations from other observatories. An increasing number of software developers are implementing AVM support in applications and an online image archive for tagged images is under development at the Spitzer Science Center. Tagging images following the AVM offers ever-increasing benefits to public-friendly imagery in all its standard forms (JPEG, TIFF, PNG). The AVM standard is one part of the Virtual Astronomy Multimedia Project (VAMP); http://www.communicatingastronomy.org

  3. Creating healthy work environments: a strategic perspective.

    Science.gov (United States)

    Adamson, Bonnie J

    2010-01-01

    Although I find Graham Lowe and Ben Chan's logic model and work environment metrics thought provoking, a healthy work environment framework must be more comprehensive and consider the addition of recommended diagnostic tools, vehicles to deliver the necessary change and a sustainability strategy that allows for the tweaking and refinement of ideas. Basic structure is required to frame and initiate an effective process, while allowing creativity and enhancements to be made by organizations as they learn. I support the construction of a suggested Canadian health sector framework for measuring the health of an organization, but I feel that organizations need to have some freedom in that design and the ability to incorporate their own indicators within the established proven drivers. Reflecting on my organization's experience with large-scale transformation efforts, I find that emotional intelligence along with formal leadership development and front-line engagement in Lean process improvement activities are essential for creating healthy work environments that produce the balanced set of outcomes listed in my hospital's Balanced Scorecard.

  4. Creating Affording Situations: Coaching through Animate Objects.

    Science.gov (United States)

    Baber, Chris; Khattab, Ahmad; Russell, Martin; Hermsdörfer, Joachim; Wing, Alan

    2017-10-11

    We explore the ways in which animate objects can be used to cue actions as part of coaching in Activities of Daily Living (ADL). In this case, changing the appearance or behavior of a physical object is intended to cue actions which are appropriate for a given context. The context is defined by the intention of the users, the state of the objects and the tasks for which these objects can be used. We present initial design prototypes and simple user trials which explore the impact of different cues on activity. It is shown that raising the handle of a jug, for example, not only cues the act of picking up the jug but also encourages use of the hand adjacent to the handle; that combinations of lights (on the objects) and auditory cues influence activity through reducing uncertainty; and that cueing can challenge pre-learned action sequences. We interpret these results in terms of the idea that the animate objects can be used to create affording situations, and discuss implications of this work to support relearning of ADL following brain damage or injury, such as might arise following a stroke.

  5. Creating Affording Situations: Coaching through Animate Objects

    Directory of Open Access Journals (Sweden)

    Chris Baber

    2017-10-01

    Full Text Available We explore the ways in which animate objects can be used to cue actions as part of coaching in Activities of Daily Living (ADL. In this case, changing the appearance or behavior of a physical object is intended to cue actions which are appropriate for a given context. The context is defined by the intention of the users, the state of the objects and the tasks for which these objects can be used. We present initial design prototypes and simple user trials which explore the impact of different cues on activity. It is shown that raising the handle of a jug, for example, not only cues the act of picking up the jug but also encourages use of the hand adjacent to the handle; that combinations of lights (on the objects and auditory cues influence activity through reducing uncertainty; and that cueing can challenge pre-learned action sequences. We interpret these results in terms of the idea that the animate objects can be used to create affording situations, and discuss implications of this work to support relearning of ADL following brain damage or injury, such as might arise following a stroke.

  6. Peculiarities of Creating Foreign Students’ Lexical Competence

    Directory of Open Access Journals (Sweden)

    Aigul Eskermesovna Sadenova

    2016-11-01

    Full Text Available Teaching of vocabulary includes several aspects of teaching content: linguistic, methodological and psychological. Teaching of vocabulary is a process which is inextricably entwined with understanding relations between new words and phonetic and grammar aspects of the language. On the one hand, simultaneous learning of two languages complicates the tasks set to the students, but, on the other hand, it simplifies the process. The complexity is in the scope of vocabulary to be remembered, as well as in the differences between grammar structures used in Russian and Kazakh. The simplification is in the fact that students are fully aware that lexical competence is formed gradually, and that every language has its lexical base required at the initial stage, and that such base is to be developed for mastering a specific language. Before identifying the methods of vocabulary semantization in the course of teaching a language to foreign students, students shall be offered active vocabulary, used to express their ideas orally and in written form, and passive vocabulary, used to perceive oral and written information. It is not possible to teach semantics only. It is necessary to create paradigmatic, syntactic and associative relations. The formation of lexical skills is connected with the solidity of vocabulary retention. In order to ensure reinforcement of the vocabulary learnt, the students shall be offered exercises contributing to the development of their skills of using vocabulary in listening, speaking, reading and writing. All above mentioned types of vocabulary semantization constitute a unified whole. Different ways of semantization are set forth for methodological purposes, to facilitate the achievement of the desired result. Translation and non-translation techniques for vocabulary presentation, as well as some tips and exercises, are given. The use of basic vocabulary at elementary and advanced levels is suggested. Certain methods for the

  7. Creating Porcine Biomedical Models Through Recombineering

    Directory of Open Access Journals (Sweden)

    Lawrence B. Schook

    2006-03-01

    Full Text Available Recent advances in genomics provide genetic information from humans and other mammals (mouse, rat, dog and primates traditionally used as models as well as new candidates (pigs and cattle. In addition, linked enabling technologies, such as transgenesis and animal cloning, provide innovative ways to design and perform experiments to dissect complex biological systems. Exploitation of genomic information overcomes the traditional need to choose naturally occurring models. Thus, investigators can utilize emerging genomic knowledge and tools to create relevant animal models. This approach is referred to as reverse genetics. In contrast to ‘forward genetics’, in which gene(s responsible for a particular phenotype are identified by positional cloning (phenotype to genotype, the ‘reverse genetics’ approach determines the function of a gene and predicts the phenotype of a cell, tissue, or organism (genotype to phenotype. The convergence of classical and reverse genetics, along with genomics, provides a working definition of a ‘genetic model’ organism (3. The recent construction of phenotypic maps defining quantitative trait loci (QTL in various domesticated species provides insights into how allelic variations contribute to phenotypic diversity. Targeted chromosomal regions are characterized by the construction of bacterial artificial chromosome (BAC contigs to isolate and characterize genes contributing towards phenotypic variation. Recombineering provides a powerful methodology to harvest genetic information responsible for phenotype. Linking recombineering with gene-targeted homologous recombination, coupled with nuclear transfer (NT technology can provide ‘clones’ of genetically modified animals.

  8. Creating a Lunar EVA Work Envelope

    Science.gov (United States)

    Griffin, Brand N.; Howard, Robert; Rajulu, Sudhakar; Smitherman, David

    2009-01-01

    A work envelope has been defined for weightless Extravehicular Activity (EVA) based on the Space Shuttle Extravehicular Mobility Unit (EMU), but there is no equivalent for planetary operations. The weightless work envelope is essential for planning all EVA tasks because it determines the location of removable parts, making sure they are within reach and visibility of the suited crew member. In addition, using the envelope positions the structural hard points for foot restraints that allow placing both hands on the job and provides a load path for reacting forces. EVA operations are always constrained by time. Tasks are carefully planned to ensure the crew has enough breathing oxygen, cooling water, and battery power. Planning first involves computers using a virtual work envelope to model tasks, next suited crew members in a simulated environment refine the tasks. For weightless operations, this process is well developed, but planetary EVA is different and no work envelope has been defined. The primary difference between weightless and planetary work envelopes is gravity. It influences anthropometry, horizontal and vertical mobility, and reaction load paths and introduces effort into doing "overhead" work. Additionally, the use of spacesuits other than the EMU, and their impacts on range of motion, must be taken into account. This paper presents the analysis leading to a concept for a planetary EVA work envelope with emphasis on lunar operations. There is some urgency in creating this concept because NASA has begun building and testing development hardware for the lunar surface, including rovers, habitats and cargo off-loading equipment. Just as with microgravity operations, a lunar EVA work envelope is needed to guide designers in the formative stages of the program with the objective of avoiding difficult and costly rework.

  9. Creating the environment for innovation and entrepreneurship

    Directory of Open Access Journals (Sweden)

    Paul M. Lane

    2016-10-01

    Full Text Available Innovation generation and diffusion have been widely acknowledged as hinging upon the complex set of institutional, social and psychological processes. The objective of the paper is to examine the need and possibilities of creating the environment for innovation and entrepreneurship in the university setting. In particular, this paper focuses on interrelationships and roles of specific groups of university members: the administration, faculty and students. The study is exploratory in character, based on observations and literature review. It starts with defining modern-day university as home of innovation emphasizing the need for interdisciplinary and interinstitutional approaches. The paper discusses the need of faculty to learn to reach across the institution and beyond to work with others, working across silos of academia and meeting with others from very different disciplines. Second it examines ways the university administration support can encourage innovation among its faculty, staff and students. Third, it draws on existing research to identify key dimensions of change. The study proposes pathways that may activate the mechanisms of climate and infrastructure for innovation. The proposed dimensions and analyzed areas of change can potentially form the foundations of a framework for universities seeking to diagnose their existing condition and use such findings to enhance the generation and diffusion of innovation. The university quest to break down the barriers and reach across the disciplines to generate innovation takes commitment which needs to be coupled with administrative change such as the reward structures lined up with the vision and changes regarding the teaching and learning practices as well as the physical environment for the classes, the class rooms and meeting spaces of students and faculty.

  10. Employee satisfaction: creating a positive work force.

    Science.gov (United States)

    Wright, M

    1998-01-01

    In the early 1990s, El Camino Hospital (ECH) streamlined its operations in order to remain competitive. In 1992, the hospital's District Board voted to turn the hospital's management over to a nonprofit company and it became an integrated delivery system (IDS). Hospital employees continued to suffer as their work and work schedules changed in ongoing efforts by the new administration to streamline. Finally, in early 1997, the IDS, Camino Healthcare, was dissolved. The director of radiology and radiation oncology services became aware of increasing employee problems, from high turnover rates and increased absenteeism, to morale and productivity issues. Employees also worried about job redesign, re-engineering and a lack of clear direction and expectations from department leadership. The director of the department created a task force to respond to the needs of staff members. With so much anger directed at department leadership, supervisory staff were not included in the task force. The task force worked first to identify rumors and innuendos and followed with a plan to resolve such issues. The second step was to agree to focus on issues that they could change and to let go of those they couldn't. They selected five priority issues or concerns. The group met weekly and made progress by replacing negative talk and attitudes with positive ones. Meanwhile, the director researched employee satisfaction issues so she would be prepared to discuss such issues and concerns with employees. She focused on a common theme, of having a personal mission or goal for one's self. She encouraged staff members to be aware of their own behavior when communicating with others. Although several informal surveys proved there was still much work to be done, there was positive response--a light at the end of the long tunnel.

  11. DOES CIVIL SOCIETY CREATE SOCIAL ENTREPRENEURS?

    Directory of Open Access Journals (Sweden)

    Gauca Oana

    2013-07-01

    Full Text Available The purpose of this paper is to analyze whether civil society itself can enhance or stimulate the creation of social entrepreneurs, by studying the traits of the civil society and the various definitions attributed to it. The main question that the paper wants to answer to is Does civil society create social entrepreneurs and the main approach used in this research paper is the theoretical one. By studying existing articles and books on the topic, the paper tries to emphasize the various dimensions that civil society can embrace, as pictured by various authors, as well as how these dimensions can relate to social entrepreneurs and the emergence of social businesses. The paper is not meant to be a breakthrough in the field, but rather to launch a question that is related to very important topics these days, social entrepreneurship, social innovation, social businesses and their connection to a very much debated topic-civil society. The paper is work-in progress and wants to stimulate research regarding the search of the sources of social entrepreneurship, in order to analyze them and better establish them as incubators for the future. It wants to be of use to whoever is researching the concepts illustrated above, as well as for those who want to get in touch with the new buzz words of the academic and entrepreneurial fields. The hereby paper stands, as previously stated, in a theoretical framework and the findings represent a mere analysis of the cause-effect relationship between the characteristics of civil society and those of social entrepreneurs. However, we are of the opinion that it can be a very good starting point for the ones interested in the domain, to analyze more sources of social entrepreneurship or further refine the answer to the question addressed in this article.

  12. Mutations in the alpha-helical region of the amino terminus of the Maize rayado fino virus capsid protein and CP:RNA ratios affect virus-like particle encapsidation of RNAs.

    Science.gov (United States)

    Natilla, Angela; Murphy, Charles; Hammond, Rosemarie W

    2015-01-22

    Viral-based nanoplatforms rely on balancing the delicate array of virus properties to optimally achieve encapsidation of foreign materials with various potential objectives. We investigated the use of Maize rayado fino virus (MRFV)-virus-like particles (VLPs) as a multifunctional nanoplatform and their potential application as protein cages. MRFV-VLPs are composed of two serologically related, carboxy co-terminal coat proteins (CP1 and CP2) which are capable of self-assembling in Nicotiana benthamiana plants into 30nm particles with T=3 symmetry. The N-terminus of CP1 was targeted for genetic modification to exploit the driving forces for VLP assembly, packaging and retention of RNA in vivo and in vitro. The N-terminus of MRFV-CP1 contains a peptide sequence of 37 amino acids which has been predicted to have an alpha-helical structure, is rich in hydrophobic amino acids, facilitates CP-RNA interactions, and is not required for self-assembly. Amino acid substitutions were introduced in the 37 amino acid N-terminus by site-directed mutagenesis and the mutant VLPs produced in plants by a Potato virus X (PVX)-based vector were tested for particle stability and RNA encapsidation. All mutant CPs resulted in production of VLPs which encapsidated non-viral RNAs, including PVX genomic and subgenomic (sg) RNAs, 18S rRNA and cellular and viral mRNAs. In addition, MRFV-VLPs encapsidated GFP mRNA when was expressed in plant cells from the pGD vector. These results suggest that RNA packaging in MRFV-VLPs is predominantly driven by electrostatic interactions between the N-terminal 37 amino acid extension of CP1 and RNA, and that the overall species concentration of RNA in the cellular pool may determine the abundance and species of the RNAs packaged into the VLPs. Furthermore, RNA encapsidation is not required for VLPs stability, VLPs formed from MRFV-CP1 were stable at temperatures up to 70°C, and can be disassembled into CP monomers, which can then reassemble in vitro into

  13. The pH Stability of Foot-and-Mouth Disease Virus Particles Is Modulated by Residues Located at the Pentameric Interface and in the N Terminus of VP1.

    Science.gov (United States)

    Caridi, Flavia; Vázquez-Calvo, Angela; Sobrino, Francisco; Martín-Acebes, Miguel A

    2015-05-01

    The picornavirus foot-and-mouth disease virus (FMDV) is the etiological agent of a highly contagious disease that affects important livestock species. The FMDV capsid is highly acid labile, and viral particles lose infectivity due to their disassembly at pH values slightly below neutrality. This acid sensitivity is related to the mechanism of viral uncoating and genome penetration from endosomes. In this study, we have analyzed the molecular basis of FMDV acid-induced disassembly by isolating and characterizing a panel of novel FMDV mutants differing in acid sensitivity. Amino acid replacements altering virion stability were preferentially distributed in two different regions of the capsid: the N terminus of VP1 and the pentameric interface. Even more, the acid labile phenotype induced by a mutation located at the pentameric interface in VP3 could be compensated by introduction of an amino acid substitution in the N terminus of VP1. These results indicate that the acid sensitivity of FMDV can be considered a multifactorial trait and that virion stability is the fine-tuned product of the interaction between residues from different capsid proteins, in particular those located within the N terminus of VP1 or close to the pentameric interface. The viral capsid protects the viral genome from environmental factors and contributes to virus dissemination and infection. Thus, understanding of the molecular mechanisms that modulate capsid stability is of interest for the basic knowledge of the biology of viruses and as a tool to improve the stability of conventional vaccines based on inactivated virions or empty capsids. Using foot-and-mouth disease virus (FMDV), which displays a capsid with extreme acid sensitivity, we have performed a genetic study to identify the molecular determinants involved in capsid stability. A panel of FMDV mutants with differential sensitivity to acidic pH was generated and characterized, and the results showed that two different regions of FMDV

  14. Creating internal culture to ensure transparency

    International Nuclear Information System (INIS)

    Hayden, E.

    2007-01-01

    Among the keys to achieving public confidence, is openness and transparency to those one serves. As a Federal regulator entrusted by the American people to protect them against the hazards of radiation, the U.S. Nuclear Regulatory Commission recognizes the need for openness and a strong 'safety culture and climate' where there is a 'safety-first focus' by its employees as well as those it regulates. For the NRC and nuclear industry, safety culture is typically the assembly of characteristics and attitudes in organisations and individuals which establishes nuclear safety as an overriding priority. Strong safety cultures include conservative decision making, strict adherence to procedures, questioning attitudes, and an environment in which employees feel free to raise safety concerns. A strong internal safety culture that is transparent to others helps the NRC to be more effective in carrying out its safety job to protect the public through its oversight of the nation's nuclear power plants and other civilian uses of nuclear energy. Creating the appropriate environment or culture and communicating NBC's contribution to safety can affect employee and ultimately public perceptions about the agency's commitment to safety in its daily activities. Where there is openness and transparency, trust and confidence are likely to follow. To assess and measure its safety culture, the NRC commissioned three independent surveys to be performed in conjunction with some use of focus groups over an 8-year period. The results identified strengths and weakness, and were compared to previous survey results as well as to other U.S. government organisations and national benchmarks. Perhaps the most surprising results came from the 2002 survey that found a third of NRC employees questioned the agency's commitment to safety, and almost half of the staff said that they did not feel it was safe to speak up in the agency. Some changes at the agency were made and the 2005 survey results showed

  15. Virus-cell fusion inhibitory activity of novel analogue peptides based on the HP (2-20) derived from N-terminus of Helicobacter pylori Ribosomal Protein L1.

    Science.gov (United States)

    Woo, Eun-Rhan; Lee, Dong Gun; Chang, Young-Su; Park, Yoonkyung; Hahm, Kyung-Soo

    2002-12-01

    HP (2-20) (AKKVFKRLEKLFSKIQNDK) is the antibacterial sequence derived from N-terminus of Helicobacter pylori Ribosomal Protein L1 (RPL1). It has a broad-spectrum microbicidal activity in vitro that is thought to be related to the membrane-disruptive properties of the peptide. Based on the putative membrane-targeted mode of action, we postulated that HP (2-20) might be possessed virus-cell fusion inhibitory activity. To develop the novel virus-cell fusion inhibitory peptides, several analogues with amino acid substitution were designed to increase or decrease only net hydrophobic region. In particular, substitution of Gln and Asp for hydrophobic amino acid, Trp at position 17 and 19 of HP (2-20) (Anal 3) caused a dramatic increase in virus-cell fusion inhibitory activity without hemolytic effect.

  16. The N Terminus of Sarcolipin Plays an Important Role in Uncoupling Sarco-endoplasmic Reticulum Ca2+-ATPase (SERCA) ATP Hydrolysis from Ca2+ Transport

    DEFF Research Database (Denmark)

    Sahoo, Sanjaya K; Shaikh, Sana A; Sopariwala, Danesh H

    2015-01-01

    to bind SERCA throughout its kinetic cycle and promotes uncoupling of Ca(2+) transport from ATP hydrolysis. To determine the structural regions of SLN that mediate uncoupling of SERCA, we employed mutagenesis and generated chimeras of PLB and SLN. In this study we demonstrate that deletion of SLN N....... Interestingly, transfer of the PLB cytosolic domain to the SLN transmembrane (TM) and luminal tail causes the chimeric protein to lose SLN-like function. Further introduction of the PLB TM region into this chimera resulted in conversion to full PLB-like function. We also found that swapping PLB N and C termini...... with those from SLN caused the resulting chimera to acquire SLN-like function. Swapping the C terminus alone was not sufficient for this conversion. These results suggest that domains can be switched between SLN and PLB without losing the ability to regulate SERCA activity; however, the resulting chimeras...

  17. Creating a Servitude to solve an encroachment Dispute: A Solution or creating another Problem?

    Directory of Open Access Journals (Sweden)

    Zsa-Zsa Temmers Boggenpoel

    2013-12-01

    Full Text Available The main focus of this note is the case of Roseveare v Katmer, Katmer v Roseveare 2013 ZAGPJHC 18, which provides an interesting (though possibly constitutionally problematic perspective to the encroachment problem. The decision in this case has opened the door for courts to create servitudes in instances where encroachments are left intact based on policy reasons. Concerning these policy reasons, the note investigates the reasonableness standard as it was applied in the case. It is argued that it is important to differentiate between the applications of reasonableness in encroachment cases and alleged nuisance disputes. The decision in this case creates the impression that courts may now order that a servitude be registered in favour of the encroacher against the affected landowner’s property. It seems as though the court had in mind the creation of a praedial servitude to justify the continued existence of the encroachment. The servitude is created by court order against the will of the affected landowner. At common law, the creation of a servitude in this respect does not exist, and the authority from which the power derives to make an order like this is not entirely clear. The court also does not provide any authority for the creation of the servitude in favour of the encroacher. Consequently, it is argued that this may have serious constitutional implications. For one, lack of authority for the deprivation that results may be unconstitutional because there is no law of general application that authorises the deprivation in terms of section 25(1. The creation of a servitude to explain the continued existence of the encroachment is not automatically included in the general discretion to replace removal with compensation. It is contended that an order that forces the affected landowner to register a servitude in favour of the encroacher to preserve the existing encroachment situation will be in conflict with section 25(1 as far as the common

  18. The amino-terminus of the hepatitis C virus (HCV p7 viroporin and its cleavage from glycoprotein E2-p7 precursor determine specific infectivity and secretion levels of HCV particle types.

    Directory of Open Access Journals (Sweden)

    Solène Denolly

    2017-12-01

    Full Text Available Viroporins are small transmembrane proteins with ion channel activities modulating properties of intracellular membranes that have diverse proviral functions. Hepatitis C virus (HCV encodes a viroporin, p7, acting during assembly, envelopment and secretion of viral particles (VP. HCV p7 is released from the viral polyprotein through cleavage at E2-p7 and p7-NS2 junctions by signal peptidase, but also exists as an E2p7 precursor, of poorly defined properties. Here, we found that ectopic p7 expression in HCVcc-infected cells reduced secretion of particle-associated E2 glycoproteins. Using biochemical assays, we show that p7 dose-dependently slows down the ER-to-Golgi traffic, leading to intracellular retention of E2, which suggested that timely E2p7 cleavage and p7 liberation are critical events to control E2 levels. By studying HCV mutants with accelerated E2p7 processing, we demonstrate that E2p7 cleavage controls E2 intracellular expression and secretion levels of nucleocapsid-free subviral particles and infectious virions. In addition, our imaging data reveal that, following p7 liberation, the amino-terminus of p7 is exposed towards the cytosol and coordinates the encounter between NS5A and NS2-based assembly sites loaded with E1E2 glycoproteins, which subsequently leads to nucleocapsid envelopment. We identify punctual mutants at p7 membrane interface that, by abrogating NS2/NS5A interaction, are defective for transmission of infectivity owing to decreased secretion of core and RNA and to increased secretion of non/partially-enveloped particles. Altogether, our results indicate that the retarded E2p7 precursor cleavage is essential to regulate the intracellular and secreted levels of E2 through p7-mediated modulation of the cell secretory pathway and to unmask critical novel assembly functions located at p7 amino-terminus.

  19. Shifting focus to create knowledge collaboratively

    Directory of Open Access Journals (Sweden)

    Hanna Voog

    2013-12-01

    the journals/editors point of view and then follow up with a more practical session on where, how and what kind of information you search in order to find relevant information about possible journals to send manuscripts to. This course design creates dynamic connections between the more classic way of academic lectures/discussions and tools that improve the PhD students' generic skills. As a result, valuable connections are being made and it is also a way of visualizing how academia works and how the culture is communicated in different ways. This includes everything from demystifying the "back-stage processes" of academia as well as finding relevant tools to navigate in a rapidly changing publication process. Both workshops and home assignments are built upon the idea of constructive alignment. The students are to focus on their respective subject field when working with the assignments, to make them matter to the individual.

  20. Difficult Sudoku Puzzles Created by Replica Exchange Monte Carlo Method

    OpenAIRE

    Watanabe, Hiroshi

    2013-01-01

    An algorithm to create difficult Sudoku puzzles is proposed. An Ising spin-glass like Hamiltonian describing difficulty of puzzles is defined, and difficult puzzles are created by minimizing the energy of the Hamiltonian. We adopt the replica exchange Monte Carlo method with simultaneous temperature adjustments to search lower energy states efficiently, and we succeed in creating a puzzle which is the world hardest ever created in our definition, to our best knowledge. (Added on Mar. 11, the ...

  1. Gender differentiation in the Bible: created and recognized

    African Journals Online (AJOL)

    This paper, however, does not discuss the issue of the treament of women in ... obvious here is that God did not create another 'adam, but he created 'issha who .... According to 1:27, God created 'adam in his image.27 Here, the singular in v. ...... Theological Interpretation of Genesis 1-3 (New York: The MacMillan Company,.

  2. Creating student awareness to improve cultural competence: creating the critical incident.

    Science.gov (United States)

    Morell, Venita W; Sharp, Penny C; Crandall, Sonia J

    2002-09-01

    Teaching medical students to recognize the need for cultural competence and accept their shortcomings in this area is a challenge. A simulated patient scenario was developed to address this challenge. The objective of the simulation is to enhance students' readiness to learn by moving them from 'unconscious incompetence' to 'conscious incompetence'. The patient scenario presents a Cherokee Indian woman with a complaint of abnormal menstrual bleeding who is resistant to gynaecologic care from male providers. A faculty member facilitates a small-group videotape review of student interviews. As students discuss their encounters, they realize they 'misdiagnose' and mishandle the interview. They are confronted by their inability to recognize cultural cues and the impact they may have on health outcomes and begin to question whether cultural beliefs are affecting the care of other patients. This simulation creates an eye-opening situation that must be handled carefully. This activity is an effective method to create awareness in students who feel they 'know all this stuff.'

  3. Haploinsufficiency of the E3 ubiquitin ligase C-terminus of heat shock cognate 70 interacting protein (CHIP produces specific behavioral impairments.

    Directory of Open Access Journals (Sweden)

    Bethann McLaughlin

    Full Text Available The multifunctional E3 ubiquitin ligase CHIP is an essential interacting partner of HSP70, which together promote the proteasomal degradation of client proteins. Acute CHIP overexpression provides neuroprotection against neurotoxic mitochondrial stress, glucocorticoids, and accumulation of toxic amyloid fragments, as well as genetic mutations in other E3 ligases, which have been shown to result in familial Parkinson's disease. These studies have created a great deal of interest in understanding CHIP activity, expression and modulation. While CHIP knockout mice have the potential to provide essential insights into the molecular control of cell fate and survival, the animals have been difficult to characterize in vivo due to severe phenotypic and behavioral dysfunction, which have thus far been poorly characterized. Therefore, in the present study we conducted a battery of neurobehavioral and physiological assays of adult CHIP heterozygotic (HET mutant mice to provide a better understanding of the functional consequence of CHIP deficiency. We found that CHIP HET mice had normal body and brain weight, body temperature, muscle tone and breathing patterns, but do have a significant elevation in baseline heart rate. Meanwhile basic behavioral screens of sensory, motor, emotional and cognitive functions were normative. We observed no alterations in performance in the elevated plus maze, light-dark preference and tail suspension assays, or two simple cognitive tasks: novel object recognition and spontaneous alternation in a Y maze. Significant deficits were found, however, when CHIP HET mice performed wire hang, inverted screen, wire maneuver, and open field tasks. Taken together, our data indicate a clear subset of behaviors that are altered at baseline in CHIP deficient animals, which will further guide whole animal studies of the effects of CHIP dysregulation on cardiac function, brain circuitry and function, and responsiveness to environmental and

  4. Applying Independent Component Analysis on Sentinel-2 Imagery to Characterize Geomorphological Responses to an Extreme Flood Event near the Non-Vegetated Río Colorado Terminus, Salar de Uyuni, Bolivia

    Directory of Open Access Journals (Sweden)

    Jiaguang Li

    2018-05-01

    Full Text Available In some internally-draining dryland basins, ephemeral river systems terminate at the margins of playas. Extreme floods can exert significant geomorphological impacts on the lower reaches of these river systems and the playas, including causing changes to flood extent, channel-floodplain morphology, and sediment dispersal. However, the characterization of these impacts using remote sensing approaches has been challenging owing to variable vegetation and cloud cover, as well as the commonly limited spatial and temporal resolution of data. Here, we use Sentinel-2 Multispectral Instrument (MSI data to investigate the flood extent, flood patterns and channel-floodplain morphodynamics resulting from an extreme flood near the non-vegetated terminus of the Río Colorado, located at the margins of the world’s largest playa (Salar de Uyuni, Bolivia. Daily maximum precipitation frequency analysis based on a 42-year record of daily precipitation data (1976 through 2017 indicates that an approximately 40-year precipitation event (40.7 mm occurred on 6 January 2017, and this was associated with an extreme flood. Sentinel-2 data acquired after this extreme flood were used to separate water bodies and land, first by using modified normalized difference water index (MNDWI, and then by subsequently applying independent component analysis (ICA on the land section of the combined pre- and post-flood images to extract flooding areas. The area around the Río Colorado terminus system was classified into three categories: water bodies, wet land, and dry land. The results are in agreement with visual assessment, with an overall accuracy of 96% and Kappa of 0.9 for water-land classification and an overall accuracy of 83% and Kappa of 0.65 for dry land-wet land classification. The flood extent mapping revealed preferential overbank flow paths on the floodplain, which were closely related to geomorphological changes. Changes included the formation and enlargement of

  5. Creating 3D gelatin phantoms for experimental evaluation in biomedicine

    Directory of Open Access Journals (Sweden)

    Stein Nils

    2015-09-01

    Full Text Available We describe and evaluate a setup to create gelatin phantoms by robotic 3D printing. Key aspects are the large workspace, reproducibility and resolution of the created phantoms. Given its soft tissue nature, the gelatin is kept fluid during inside the system and we present parameters for additive printing of homogeneous, solid objects. The results indicate that 3D printing of gelatin can be an alternative for quickly creating larger soft tissue phantoms without the need for casting a mold.

  6. CREATING A CORPORATE ENTREPRENEURSHIP STRATEGY FOR COMPETITIVE ADVANTAGE

    OpenAIRE

    ENGİNOĞLU, Didem; ARIKAN, Cenk Laçin

    2016-01-01

    Current competitive environment is rapidly changing. In today’s business environment, organizations are having an increasingly difficult time in creating competitive advantages. The main reason for this is the ease in contemporary business life for organizations to reach the same or very similar resources. Firms need innovation to create and sustain success and effectiveness. In such a highly competitive business life, the importance of creating competitive advantages for organizations based ...

  7. Creating visual guidelines for a Finnish fashion brand

    OpenAIRE

    Morozova, Elizaveta

    2017-01-01

    Visual communication is an essential part of branding. Visual elements like logo, brand colours, and typography create associations with a brand and make it distinctive. Graphic design is a tool for visual communication that allows creating aesthetic and functional visual elements that convey some messages. The client of this thesis is a Finnish fashion company that does not have clear and consistent visual image, and the goal is to create it based on company’s values and desired image...

  8. Creating a Curriculum for Training Health Profession Faculty Leaders

    National Research Council Canada - National Science Library

    Mitchell, Pamela H; Robins, Lynne S; Schaad, Dotiglas

    2005-01-01

    Objectives: An interprofessional, collaborative group of educators, patient safety officers, and Federal program directors teamed up to create an integrated, patient safety-centered curriculum for the education...

  9. In vitro and in vivo Analysis of the Binding of the C Terminus of the HDL Receptor Scavenger Receptor Class B type I (SR-BI) to the PDZ1 Domain of its Cytoplasmic Adaptor Protein PDZK1

    Energy Technology Data Exchange (ETDEWEB)

    O Kocher; G Birrane; K Tsukamoto; S Fenske; A Yesilaltay; R Pal; K Daniels; J Ladias; M Krieger

    2011-12-31

    The PDZ1 domain of the four PDZ domain-containing protein PDZK1 has been reported to bind the C terminus of the HDL receptor scavenger receptor class B, type I (SR-BI), and to control hepatic SR-BI expression and function. We generated wild-type (WT) and mutant murine PDZ1 domains, the mutants bearing single amino acid substitutions in their carboxylate binding loop (Lys(14)-Xaa(4)-Asn(19)-Tyr-Gly-Phe-Phe-Leu(24)), and measured their binding affinity for a 7-residue peptide corresponding to the C terminus of SR-BI ((503)VLQEAKL(509)). The Y20A and G21Y substitutions abrogated all binding activity. Surprisingly, binding affinities (K(d)) of the K14A and F22A mutants were 3.2 and 4.0 ?M, respectively, similar to 2.6 ?M measured for the WT PDZ1. To understand these findings, we determined the high resolution structure of WT PDZ1 bound to a 5-residue sequence from the C-terminal SR-BI ((505)QEAKL(509)) using x-ray crystallography. In addition, we incorporated the K14A and Y20A substitutions into full-length PDZK1 liver-specific transgenes and expressed them in WT and PDZK1 knock-out mice. In WT mice, the transgenes did not alter endogenous hepatic SR-BI protein expression (intracellular distribution or amount) or lipoprotein metabolism (total plasma cholesterol, lipoprotein size distribution). In PDZK1 knock-out mice, as expected, the K14A mutant behaved like wild-type PDZK1 and completely corrected their hepatic SR-BI and plasma lipoprotein abnormalities. Unexpectedly, the 10-20-fold overexpressed Y20A mutant also substantially, but not completely, corrected these abnormalities. The results suggest that there may be an additional site(s) within PDZK1 that bind(s) SR-BI and mediate(s) productive SR-BI-PDZK1 interaction previously attributed exclusively to the canonical binding of the C-terminal SR-BI to PDZ1.

  10. Net-Zero Building Technologies Create Substantial Energy Savings -

    Science.gov (United States)

    only an estimated 1% of commercial buildings are built to net-zero energy criteria. One reason for this Continuum Magazine | NREL Net-Zero Building Technologies Create Substantial Energy Savings Net -Zero Building Technologies Create Substantial Energy Savings Researchers work to package and share step

  11. Talent Management for Creating a Performance Work Environment ...

    African Journals Online (AJOL)

    This article examines the extent to which talent management can contribute towards creating a performance work environment (PWE) that can enhance sustainable talent identifi cation and development in the public service. The literature analysis results reveal that talent management is essential in creating a PWE in the ...

  12. Creating Futures through Participation and open-ended experiments

    DEFF Research Database (Denmark)

    Husted, Mia; Bladt, Mette; Tofteng, Ditte Maria Børglum

    This workshop will engage participants in a process that enables ‘ordinary people’ to take responsibility for creating new futures and to bring about new anticipations for democratic developments of local and societal issues. Founded in the Critical Utopian Action Research Tradition, the workshop...... will engage participants in a brief version of a Future Creating Workshop....

  13. 5 CFR 9701.372 - Creating initial pay ranges.

    Science.gov (United States)

    2010-01-01

    ... 5 Administrative Personnel 3 2010-01-01 2010-01-01 false Creating initial pay ranges. 9701.372... HUMAN RESOURCES MANAGEMENT SYSTEM Pay and Pay Administration Transitional Provisions § 9701.372 Creating initial pay ranges. (a) DHS must, after coordination with OPM, set the initial band rate ranges for the...

  14. Creating Cartoons to Promote Leaderships Skills and Explore Leadership Qualities

    Science.gov (United States)

    Smith, Latisha L.; Clausen, Courtney K.; Teske, Jolene K.; Ghayoorrad, Maryam; Gray, Phyllis; Al Subia, Sukainah; Atwood-Blaine, Dana; Rule, Audrey C.

    2015-01-01

    This document describes a strategy for increasing student leadership and creativity skills through the creation of cartoons. Creating cartoons engages students in divergent thinking and cognitive processes, such as perception, recall, and mental processing. When students create cartoons focused on a particular topic, they are making connections to…

  15. Fundamental Characteristics of Incentive Streams Created by Legal Systems

    NARCIS (Netherlands)

    Dari Mattiacci, G.

    2002-01-01

    The law shapes people’s behaviour by creating incentives. For example, tort law induces motorists to drive carefully by making them pay compensation for the accidents they may cause. This study analyses the way the law can create incentives in those cases in which the courts or the administrative

  16. Mandatory Parent Education Programs Can Create Positive Youth Sport Experiences

    Science.gov (United States)

    Christofferson, Jennifer; Strand, Bradford

    2016-01-01

    Youth sport leaders must not ignore the influence parents have on creating a positive developmental experience for young athletes. Therefore, expectations involving parental involvement and conduct must be addressed prior to athletes' participation. This article aims to examine the importance of creating mandatory parental training programs for…

  17. Utilizing Educational Corporate Culture To Create a Quality School.

    Science.gov (United States)

    Osborne, Bill

    Strategies for utilizing educational corporate culture to create a quality school are presented in this paper, which argues that the understanding of the shared belief system of organizational members is crucial to the process. Creating a quality school entails moving from a "teach the process" oriented model to one that internalizes the…

  18. Elimination of polarity in the carotenoid terminus promotes the exposure of B850-binding sites (Tyr 44, 45) and ANS-mediated energy transfer in LH2 complexes of Rhodobacter sphaeroides.

    Science.gov (United States)

    Liu, Yuan; Wu, Yongqiang; Xu, Chunhe

    2004-12-10

    Carotenoids in the peripheral light-harvesting complexes (LH2) of the green mutant (GM309) of Rhodobacter sphaeroides were identified as containing neurosporenes, which lack the polar CH(3)O group, compared to spheroidenes in native-LH2 of R. sphaeroides 601. After LH2 complexes were treated with 1-anilino-8-naphthalene sulfonate (ANS), new energy transfer pathways from ANS or tryptophan to carotenoids were discovered in both native- and GM309-LH2. The carotenoid fluorescence intensity of GM309-LH2 was greater than that of native-LH2 when bound with ANS, suggesting that the elimination of polarity in the neurosporene increases the energy transfer from ANS to carotenoid. The fact that two alpha-tyrosines (alpha-Tyr 44, 45, B850-binding sites) in each alpha-apoprotein of GM309-LH2 were more easily modified than those of native-LH2 by N-acetylimidazole (NAI) indicates that the elimination of polarity in the neurosporene terminus increases the exposure of these sites to solution.

  19. The Chikungunya Virus Capsid Protein Contains Linear B Cell Epitopes in the N- and C-Terminal Regions that are Dependent on an Intact C-Terminus for Antibody Recognition

    Directory of Open Access Journals (Sweden)

    Lucas Y. H. Goh

    2015-06-01

    Full Text Available Chikungunya virus (CHIKV is an arthropod-borne agent that causes severe arthritic disease in humans and is considered a serious health threat in areas where competent mosquito vectors are prevalent. CHIKV has recently been responsible for several millions of cases of disease, involving over 40 countries. The recent re-emergence of CHIKV and its potential threat to human health has stimulated interest in better understanding of the biology and pathogenesis of the virus, and requirement for improved treatment, prevention and control measures. In this study, we mapped the binding sites of a panel of eleven monoclonal antibodies (mAbs previously generated towards the capsid protein (CP of CHIKV. Using N- and C-terminally truncated recombinant forms of the CHIKV CP, two putative binding regions, between residues 1–35 and 140–210, were identified. Competitive binding also revealed that five of the CP-specific mAbs recognized a series of overlapping epitopes in the latter domain. We also identified a smaller, N-terminally truncated product of native CP that may represent an alternative translation product of the CHIKV 26S RNA and have potential functional significance during CHIKV replication. Our data also provides evidence that the C-terminus of CP is required for authentic antigenic structure of CP. This study shows that these anti-CP mAbs will be valuable research tools for further investigating the structure and function of the CHIKV CP.

  20. Online by design the essentials of creating information literacy courses

    CERN Document Server

    Mery, Yvonne

    2014-01-01

    As online instruction becomes more popular, librarians will want to develop the knowledge and skills needed to create an effective online information literacy course. Online by Design: The Essentials of Creating Information Literacy Courses will guide librarians as they go through the process of designing, developing, and delivering online information literacy courses. Yvonne Mery & Jill Newby offer proven techniques and tips for creating quality online courses that are engaging and effective. This handbook is perfect for instruction librarians who are interesting in developing new courses or

  1. 3D game environments create professional 3D game worlds

    CERN Document Server

    Ahearn, Luke

    2008-01-01

    The ultimate resource to help you create triple-A quality art for a variety of game worlds; 3D Game Environments offers detailed tutorials on creating 3D models, applying 2D art to 3D models, and clear concise advice on issues of efficiency and optimization for a 3D game engine. Using Photoshop and 3ds Max as his primary tools, Luke Ahearn explains how to create realistic textures from photo source and uses a variety of techniques to portray dynamic and believable game worlds.From a modern city to a steamy jungle, learn about the planning and technological considerations for 3D modelin

  2. Creating a Podcast/Vodcast: A How-To Approach

    Science.gov (United States)

    Petersen, C. C.

    2011-09-01

    Creating podcasts and vodcasts is a wonderful way to share news of science research. Public affairs officers use them to reveal the latest discoveries done by scientists in their institutions. Educators can offer podcast/vodcast creation for students who want a unique way to demonstrate their mastery of science topics. Anyone with a computer and a USB microphone can create a podcast. To do a vodcast, you also need a digital video camera and video editing software. This session focused mainly on creating a podcast - writing the script and recording the soundtrack. Attendees also did a short activity to learn to write effective narrative copy for a podcast/vodcast.

  3. Resonant fields created by spiral electric currents in Tokamaks

    International Nuclear Information System (INIS)

    Fernandes, A.S.; Caldas, I.L.

    1985-01-01

    The influence of the resonant magnetic perturbations, created by electric currents in spirals, on the plasma confinement in a tokamak with circular section and large aspect ratio is investigated. These perturbations create magnetic islands around the rational magnetic surface which has the helicity of the helicoidal currents. The intensities of these currents are calculated in order to the magnetic islands reach the limiter or others rational surfaces, what could provoke the plasma disrupture. The electric current intensities are estimated, in two spiral sets with different helicities, which create a predominantly stocastic region among the rational magnetic surfaces with these helicities. (L.C.) [pt

  4. Retreating academics: creating spaces for the scholarship of ...

    African Journals Online (AJOL)

    , this paper explores particular spaces created to support academic engagement in the scholarship of teaching and learning: the space of writing retreats. The metaphor of 'tapestry' is used to capture the development of a complex conceptual ...

  5. Creating a panorama of the heart with digital images.

    Science.gov (United States)

    Rosebrock, L

    2000-01-01

    Digital imaging offers new opportunities still being discovered by users. This article describes a technique that was created using a digital camera to photograph the entire surface of a rat heart. The technique may have other applications as well.

  6. Creating a collage: a meaningful interactive classroom strategy.

    Science.gov (United States)

    Seymour, R J

    1995-01-01

    How can students be helped to make personal meaning of their nursing knowledge? The author describes one strategy, that of creating a collage, that promoted interactive learning and the making of meaning.

  7. Creating Simple Admin Tools Using Info*Engine and Java

    Science.gov (United States)

    Jones, Corey; Kapatos, Dennis; Skradski, Cory; Felkins, J. D.

    2012-01-01

    PTC has provided a simple way to dynamically interact with Windchill using Info*Engine. This presentation will describe how to create a simple Info*Engine Tasks capable of saving Windchill 10.0 administration of tedious work.

  8. DES Science Portal: II- Creating Science-Ready Catalogs

    Energy Technology Data Exchange (ETDEWEB)

    Fausti Neto, Angelo; et al.

    2017-08-18

    We present a novel approach for creating science-ready catalogs through a software infrastructure developed for the Dark Energy Survey (DES). We integrate the data products released by the DES Data Management and additional products created by the DES collaboration in an environment known as DES Science Portal. Each step involved in the creation of a science-ready catalog is recorded in a relational database and can be recovered at any time. We describe how the DES Science Portal automates the creation and characterization of lightweight catalogs for DES Year 1 Annual Release, and show its flexibility in creating multiple catalogs with different inputs and configurations. Finally, we discuss the advantages of this infrastructure for large surveys such as DES and the Large Synoptic Survey Telescope. The capability of creating science-ready catalogs efficiently and with full control of the inputs and configurations used is an important asset for supporting science analysis using data from large astronomical surveys.

  9. Creating an adaptive healing room for neurology patients

    NARCIS (Netherlands)

    Daemen, E.M.L; Behere, S.; Cuppen, R.P.G.; Facey, J.; Flinsenberg, I.C.M.; Loenen, van E.J.; Rajae-Joordens, R.J.E.

    2013-01-01

    Informed by the experiences of patients and medical staff, Philips has designed a room for neurology patients that retains a clinical feel when treatment is being carried out, but can flex to create a moretailored, personal ambiende.

  10. Future Combat Systems (FCS) Creates Cannon and Mortar Synergy

    National Research Council Canada - National Science Library

    Beard, Kirby; James, Jeff; Tolbert, Vincent J

    2008-01-01

    .... The NLOS-C is one of the eight MGVs. Program Manager FCS (Brigade Combat Team (PM FCS(BCT)) is leveraging previous and current research and development efforts to create synergy between cannons and mortars, without duplication of effort...

  11. Communicating Disparity: How Social Design can create Public ...

    African Journals Online (AJOL)

    chrischisoni

    Key words: Inequality; communication; social design; storytelling ... created unevenness, which can destroy dignity, self-confidence and esteem. .... divides, which involves gaps between groups in awareness, adoption, skills, devices, use and.

  12. Creating a Database for Test Items in National Examinations (pp ...

    African Journals Online (AJOL)

    Nekky Umera

    different programmers create files and application programs over a long period. .... In theory or essay questions, alternative methods of solving problems are explored and ... Unworthy items are those that do not focus on the central concept or.

  13. Kids Create Healthy Comics | NIH MedlinePlus the Magazine

    Science.gov (United States)

    ... School Students Using Medline Plus Kids Create Healthy Comics Past Issues / Fall 2015 Table of Contents Fresh, ... use of reliable health information resources." The Four Comic Books Are: The Expert Investigator explores the impact ...

  14. Control of invasive plant species in created wetlands.

    Science.gov (United States)

    1994-01-01

    The Virginia Department of Transportation (VDOT) has been using a herbicide in the form of glyphosate for the control of unwanted species in created wetlands. Results with this particular herbicide have been somewhat mixed. It was requested that the ...

  15. Creating sustainable learning environments in schools by means of ...

    African Journals Online (AJOL)

    Creating sustainable learning environments in schools by means of strategic ... be addressed by means of proper strategic planning of the education system as such ... The authors who are academics at a university and who are specializing in ...

  16. Creating photo-realistic works in a 3D scene using layers styles to create an animation

    Science.gov (United States)

    Avramescu, A. M.

    2015-11-01

    Creating realist objects in a 3D scene is not an easy work. We have to be very careful to make the creation very detailed. If we don't know how to make these photo-realistic works, by using the techniques and a good reference photo we can create an amazing amount of detail and realism. For example, in this article there are some of these detailed methods from which we can learn the techniques necessary to make beautiful and realistic objects in a scene. More precisely, in this paper, we present how to create a 3D animated scene, mainly using the Pen Tool and Blending Options. Indeed, this work is based on teaching some simple ways of using the Layer Styles to create some great shadows, lights, textures and a realistic sense of 3 Dimension. The present work involves also showing how some interesting ways of using the illuminating and rendering options can create a realistic effect in a scene. Moreover, this article shows how to create photo realistic 3D models from a digital image. The present work proposes to present how to use Illustrator paths, texturing, basic lighting and rendering, how to apply textures and how to parent the building and objects components. We also propose to use this proposition to recreate smaller details or 3D objects from a 2D image. After a critic art stage, we are able now to present in this paper the architecture of a design method that proposes to create an animation. The aim is to create a conceptual and methodological tutorial to address this issue both scientifically and in practice. This objective also includes proposing, on strong scientific basis, a model that gives the possibility of a better understanding of the techniques necessary to create a realistic animation.

  17. Creating Practitioners of Design for Quality through Education

    DEFF Research Database (Denmark)

    Robotham, Antony John

    1998-01-01

    The above paper described my experiences of teaching engineering students to be able to create products of the highest design quality. The paper identifies the need to have a good understanding of the needs of the customer and to be able to create products, which have a range of quality attributes...... would be of higher quality and provide greater customer satisfaction if designers were continually aware of all life-phase needs....

  18. Use FOMO to Create Urgency in the Booking Process

    OpenAIRE

    2017-01-01

    Create a sense of urgency and “fear of missing out” (FOMO) to nudge people to book faster. People feel more compelled to buy if they believe they are about to miss out on a limited-time offer or a good deal. This type of urgency is about creating “scarcity value,” a concept that OTAs have mastered and implemented systematically on their OTA websites to stimulate purchase behavior.

  19. Management challenges in creating value from business analytics

    OpenAIRE

    Vidgen, Richard; Shaw, S.; Grant, D.G.

    2017-01-01

    The popularity of big data and business analytics has increased tremendously in the last decade and a key challenge for organizations is in understanding how to leverage them to create business value. However, while the literature acknowledges the importance of these topics little work has addressed them from the organization’s point of view. This paper investigates the challenges faced by organizational managers seeking to become more data and information-driven in order to create value. Emp...

  20. Soft Systems Methodology Embedded in Organizational Knowledge-Creating Process

    OpenAIRE

    Yoshida, Taketoshi

    2005-01-01

    We clarify the role of tacit knowing in the soft systems methodology. For the purpose we investigate the basic structure of its seven-stage model, while embedding the soft systems methodology in organizational knowledge-creating process. This leads to the introduction of concept creation to the methodology. This changes the basic shape of the soft systems methodology from learning cycle to organizational knowledge-creating spiral where concept creation is the key point.