Sample records for raphe terminus creating

  1. Median raphe cyst: report of two cases. (United States)

    Kumar, Piyush; Das, Anupam; Savant, Sushil S; Barkat, Rizwana


    Median raphe cysts are rare congenital lesions ofthe male genitalia that occur as a result of alteredembryologic development. We report two such casesof median raphe cysts in the pediatric age group. Inaddition, we review the literature.

  2. Activity of Raphé Serotonergic Neurons Controls Emotional Behaviors

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    Anne Teissier


    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. Dissociations between the effects of LSD on behavior and raphe unit activity in freely moving cats. (United States)

    Trulson, M E; Jacobs, B L


    The hypothesis that the action of hallucinogenic drugs is mediated by a depression of the activity of brain serotonergic (raphe) neurons was tested by examining the behavioral effects of d-lysergic acid diethylamide (LSD) while studying the activity of raphe neurons in freely moving cats. Although the results provide general support for the hypothesis, there were several important dissociations. (i) Low doses of LSD produced only small decreases in raphe unit activity but significant behavoiral changes; (ii) LSD-induced behavioral changes outlasted the depression of raphe unit activity; and (iii) raphe neurons were at least as responsive to LSD during tolerance as they were in the nontolerant condition.

  4. Tidewater terminus tug-of-war (United States)

    Bartholomaus, T. C.; Larsen, C. F.; O'Neel, S.; West, M. E.


    When a glacier terminus recedes, not only does the glacier lose the ice between the former and present terminus, but the terminal reach of the glacier can steepen, causing ice flow out of the glacier interior increases. The increased flow will continue, thinning the glacier, until the glacier geometry and ice flow reach a new equilibrium. Yahtse Glacier is an advancing tidewater glacier on the Gulf of Alaska coast. To better understand the controls on its terminus position, we use a suite of seismic, geodetic and oceanographic data. Both calving and submarine melt contribute to frontal ablation, however, at Yahtse Glacier the ice is too fractured to support undercutting below the water line, nor does a persistent submarine toe develop. Thus the terminus retreats as fast as subaerial calving occurs. Previous work at Yahtse Glacier demonstrated that locally recorded seismic events between 1 and 5 Hz are predominantly the result of subaerial iceberg calving. Therefore, we use seismicity as a proxy for the frontal ablation rate. We measure the near-terminus glacier velocity with oblique photogrammetry, calibrated with ~10 day intervals of surveyed ice velocity. These methods reveal an annually-averaged terminus velocity of 6.9 km/yr. The frontal ablation rate and the terminus ice velocity are nearly in phase and reach maximum values twice per year: in the spring and fall. Integrating the difference between frontal ablation rate and terminus ice velocity reveals a pattern of terminus positions with a single annual cycle, quite similar to that which we observe in the field. GPS measurements 10 km from the terminus indicate that ice velocities peak in May and decrease through the summer. Oceanographic measurements show that near-shore surface water temperatures in the Gulf of Alaska are greatest in the fall. We suggest that the spring peak in terminus velocity is set by higher rates of ice delivery from up-glacier; calving rate increases in a compensatory way, to nearly

  5. Activation of raphe nuclei triggers rapid and distinct effects on parallel olfactory bulb output channels. (United States)

    Kapoor, Vikrant; Provost, Allison C; Agarwal, Prateek; Murthy, Venkatesh N


    The serotonergic raphe nuclei are involved in regulating brain states over timescales of minutes and hours. We examined more rapid effects of raphe activation on two classes of principal neurons in the mouse olfactory bulb, mitral and tufted cells, which send olfactory information to distinct targets. Brief stimulation of the raphe nuclei led to excitation of tufted cells at rest and potentiation of their odor responses. While mitral cells at rest were also excited by raphe activation, their odor responses were bidirectionally modulated, leading to improved pattern separation of odors. In vitro whole-cell recordings revealed that specific optogenetic activation of raphe axons affected bulbar neurons through dual release of serotonin and glutamate. Therefore, the raphe nuclei, in addition to their role in neuromodulation of brain states, are also involved in fast, sub-second top-down modulation similar to cortical feedback. This modulation can selectively and differentially sensitize or decorrelate distinct output channels.

  6. The interaction between the locus coeruleus and dorsal raphe nucleus studied with dual-probe microdialysis

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    Pudovkina, OL; Cremers, TIFH; Westerink, BHC


    The interaction between the locus coeruleus and dorsal raphe nucleus was investigated by means of dual-probe microdialysis in conscious rats. The release of noradrenaline and 5-hydroxytryptamine (5-HT) after inhibition or stimulation of locus cocruleus and dorsal raphe activity was sampled in both

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

    Indian Academy of Sciences (India)

    Home; Journals; Journal of Earth System Science; Volume 125; Issue 4. Geomorphic observations from southwestern terminus of Palghat Gap, south India and their tectonic implications. Yogendra Singh Biju John G P Ganapathy Abhilash Abhilash George S Harisanth K S Divyalakshmi Sreekumari Kesavan. Volume 125 ...

  8. A dynamic physical characterization of the receding Mendenhall Glacier lake front terminus Juneau, Alaska (United States)

    Connor, C. L.; Fatland, D. R.; Heavner, M.; Korzen, N.; Galbraith, J.; Sauer, D.; Hood, E. W.


    Extrapolation of 2000-2009 GPS results from terminus position surveys of the Mendenhall Glacier near Juneau, Alaska suggests that the lake front glacier terminus will no longer be in contact with proglacial Mendenhall Lake by July 2011. Meteorologic stations located near the glacier terminus at 44m asl, on the glacier surface at 430m (Northstar Camp), and at 1569m near the Mendenhall-Taku Glacier ice divide, provide data from rainfall events and temperature variation which contribute to glacier velocity and ultimately ice mass transfer to the lower glacier. Mendenhall weather data in combination with wind direction, wind velocity, and lake water temperature profiles (0-40m) and bathymetric surveys in 2009 provide detailed information about the physical conditions of the glacier and lake which are also captured visually by hourly and 30 second image records of the glacier terminus. Cameras are located at 500m from the terminus on bedrock and at ~2km from the terminus at the USFS Mendenhall Glacier Visitor Center roof. Ice berg motions and their changing positions in Mendenhall Lake can be used to create a gyre model for lake circulation. Summer 2009 lake water column temperature profiles collected at 15 minute intervals can also be linked with met station data, and USGS discharge data for the Mendenhall River to identify subglacial meltwater discharge events into the lake. We present here a synthetic view of these sensor data to evaluate what can be inferred and what remains mysterious concerning Mendenhall Glacier recession. Webcam photo Mendenhall Glacier Terminus 01-Sept-2009 10:02 am

  9. Dorsal Raphe Dopamine Neurons Represent the Experience of Social Isolation (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.


    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

  10. Effects of Hydro Alcoholic Extraction of Valeriana on Astrocyte Raphe Magnus in Adult Rats

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    sajad Hatami joni


    Conclusion: Oral administration of hydro alcoholic extract of valerian increases astrocytes number and decreases their size in nucleus of raphe Magna, which indicated the effect of this extraction on proliferation of astrocytes increasing.

  11. Pontine-ventral respiratory column interactions through raphe circuits detected using multi-array spike train recordings. (United States)

    Nuding, Sarah C; Segers, Lauren S; Baekey, David M; Dick, Thomas E; Solomon, Irene C; Shannon, Roger; Morris, Kendall F; Lindsey, Bruce G


    Recently, Segers et al. identified functional connectivity between the ventrolateral respiratory column (VRC) and the pontine respiratory group (PRG). The apparent sparseness of detected paucisynaptic interactions motivated consideration of other potential functional pathways between these two regions. We report here evidence for "indirect" serial functional linkages between the PRG and VRC via intermediary brain stem midline raphé neurons. Arrays of microelectrodes were used to record sets of spike trains from a total of 145 PRG, 282 VRC, and 340 midline neurons in 11 decerebrate, vagotomized, neuromuscularly blocked, ventilated cats. Spike trains of 13,843 pairs of neurons that included at least one raphé cell were screened for respiratory modulation and short-time scale correlations. Significant correlogram features were detected in 7.2% of raphé-raphé (291/4,021), 4.3% of VRC-raphé (292/6,755), and 4.0% of the PRG-raphé (124/3,067) neuron pairs. Central peaks indicative of shared influences were the most common feature in correlations between pairs of raphé neurons, whereas correlated raphé-PRG and raphé-VRC neuron pairs displayed predominantly offset peaks and troughs, features suggesting a paucisynaptic influence of one neuron on the other. Overall, offset correlogram features provided evidence for 33 VRC-to-raphé-to-PRG and 45 PRG-to-raphé-to-VRC correlational linkage chains with one or two intermediate raphé neurons. The results support a respiratory network architecture with parallel VRC-to-PRG and PRG-to-VRC links operating through intervening midline circuits, and suggest that raphé neurons contribute to the respiratory modulation of PRG neurons and shape the respiratory motor pattern through coordinated divergent actions on both the PRG and VRC.


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    G. R. Hassanzadeh G. Behzadi


    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.

  13. Central and peripheral chemoreceptors evoke distinct responses in simultaneously recorded neurons of the raphé-pontomedullary respiratory network


    Nuding, Sarah C.; Segers, Lauren S.; Shannon, Roger; O'Connor, Russell; Morris, Kendall F.; Lindsey, Bruce G.


    The brainstem network for generating and modulating the respiratory motor pattern includes neurons of the medullary ventrolateral respiratory column (VRC), dorsolateral pons (PRG) and raphé nuclei. Midline raphé neurons are proposed to be elements of a distributed brainstem system of central chemoreceptors, as well as modulators of central chemoreceptors at other sites, including the retrotrapezoid nucleus. Stimulation of the raphé system or peripheral chemoreceptors can induce a long-term fa...

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

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    Blackshear, M.A.; Sanders-Bush, E. (Vanderbilt Univ., Nashville, TN (USA). School of Medicine); Steranka, L.R. (Indiana University, Northwest Center for Medical Education, Gary, IN, USA)


    These studies confirm and extend the recent work suggesting that (/sup 3/H)lysergic acid diethylamide (LSD) labels two distinct binding sites in rat brain resembling serotonin (5HT) receptors. Although Scatchard analyses of (/sup 3/H)LSD binding to membranes prepared from cortex/hippocampus were linear, the heterogeneity of the (/sup 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/sub 1/ and 5HT/sub 2/ respectively. Regional analyses showed, that in the frontal cortex, the density of the 5HT/sub 2/ site was slightly greater than the 5HT/sub 1/ site whereas the 5HT/sub 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.

  15. Nicotinic modulation of serotonergic activity in the dorsal raphe nucleus. (United States)

    Hernandez-Lopez, Salvador; Garduño, Julieta; Mihailescu, Stefan


    Cholinergic signaling mediated by nicotinic receptors has been associated to a large number of physiological and behavioral processes such as learning, memory, attention, food-intake and mood disorders. Although it is well established that many nicotinic actions are mediated through an increase in serotonin (5-HT) release, the physiological mechanisms by which nicotine produces these effects are still unclear. The dorsal raphe nucleus (DRN) contains the major amount of 5-HT neurons projecting to different parts of the brain. DRN also contains nicotinic acetylcholine receptors (nAChRs) located at somatic and presynaptic elements. Nicotine produces both inhibitory and excitatory effects on different subpopulations of 5-HT DRN neurons. In this review, we describe the presynaptic and postsynaptic mechanisms by which nicotine increases the excitability of DRN neurons as well as the subtypes of nAChRs involved. We also describe the inhibitory effects of nicotine and the role of 5-HT1A receptors in this effect. These nicotinic actions modulate the activity of different neuronal subpopulations in the DRN, changing the 5-HT tone in the brain areas where these groups of neurons project. Some of the physiological implications of nicotine-induced 5-HT release are discussed.

  16. Glaciological and marine geological controls on terminus dynamics of Hubbard Glacier, southeast Alaska (United States)

    Stearns, L. A.; Hamilton, G. S.; van der Veen, C. J.; Finnegan, D. C.; O'Neel, S.; Scheick, J. B.; Lawson, D. E.


    Hubbard Glacier, located in southeast Alaska, is the world's largest nonpolar tidewater glacier. It has been steadily advancing since it was first mapped in 1895; occasionally, the advance creates an ice or sediment dam that blocks a tributary fjord (Russell Fiord). The sustained advance raises the probability of long-term closure in the near future, which will strongly impact the ecosystem of Russell Fiord and the nearby community of Yakutat. Here, we examine a 43 year record of flow speeds and terminus position to understand the large-scale dynamics of Hubbard Glacier. Our long-term record shows that the rate of terminus advance has increased slightly since 1895, with the exception of a slowed advance between approximately 1972 and 1984. The short-lived closure events in 1986 and 2002 were not initiated by perturbations in ice velocity or environmental forcings but were likely due to fluctuations in sedimentation patterns at the terminus. This study points to the significance of a coupled system where short-term velocity fluctuations and morainal shoal development control tidewater glacier terminus position.

  17. Glaciological and marine geological controls on terminus dynamics of Hubbard Glacier, southeast Alaska (United States)

    Stearns, Leigh A.; Hamilton, Gordon S.; van der Veen, C. J.; Finnegan, D. C.; O'Neel, Shad; Scheick, J. B.; Lawson, D. E.


    Hubbard Glacier, located in southeast Alaska, is the world's largest non-polar tidewater glacier. It has been steadily advancing since it was first mapped in 1895; occasionally, the advance creates an ice or sediment dam that blocks a tributary fjord (Russell Fiord). The sustained advance raises the probability of long-term closure in the near-future, which will strongly impact the ecosystem of Russell Fiord and the nearby community of Yakutat. Here, we examine a 43-year record of flow speeds and terminus position to understand the large-scale dynamics of Hubbard Glacier. Our long-term record shows that the rate of terminus advance has increased slightly since 1895, with the exception of a slowed advance between approximately 1972 and 1984. The short-lived closure events in 1986 and 2002 were not initiated by perturbations in ice velocity or environmental forcings, but were likely due to fluctuations in sedimentation patterns at the terminus. This study points to the significance of a coupled system where short-term velocity fluctuations and morainal shoal development control tidewater glacier terminus position.

  18. Compliance crash testing of the Type 60K terminus. (United States)


    Crash testing for compliance with NCHRP Report 350 was performed on a Type 60K terminus. The Type 60K terminus was : comprised of Type 60K portable concrete barrier (TL-3 approved) anchored to Type 60 concrete barrier at one end but free at the : oth...

  19. Serotoninergic dorsal raphe neurons possess functional postsynaptic nicotinic acetylcholine receptors. (United States)

    Galindo-Charles, Luis; Hernandez-Lopez, Salvador; Galarraga, Elvira; Tapia, Dagoberto; Bargas, José; Garduño, Julieta; Frías-Dominguez, Carmen; Drucker-Colin, René; Mihailescu, Stefan


    Very few neurons in the telencephalon have been shown to express functional postsynaptic nicotinic acetylcholine receptors (nAChRs), among them, the noradrenergic and dopaminergic neurons. However, there is no evidence for postsynaptic nAChRs on serotonergic neurons. In this study, we asked if functional nAChRs are present in serotonergic (5-HT) and nonserotonergic (non-5-HT) neurons of the dorsal raphe nucleus (DRN). In rat midbrain slices, field stimulation at the tegmental pedunculopontine (PPT) nucleus evoked postsynaptic currents (eEPSCs) with different components in DRN neurons. After blocking the glutamatergic and GABAergic components, the remaining eEPSCs were blocked by mecamylamine and reduced by either the selective alpha7 nAChR antagonist methyllycaconitine (MLA) or the selective alpha4beta2 nAChR antagonist dihydro-beta-eritroidine (DHbetaE). Simultaneous addition of MLA and DHbetaE blocked all eEPSCs. Integrity of the PPT-DRN pathway was assessed by both anterograde biocytin tracing and antidromic stimulation from the DRN. Inward currents evoked by the direct application of acetylcholine (ACh), in the presence of atropine and tetrodotoxin, consisted of two kinetically different currents: one was blocked by MLA and the other by DHbetaE; in both 5-HT and non-5-HT DR neurons. Analysis of spontaneous (sEPSCs) and evoked (eEPSCs) synaptic events led to the conclusion that nAChRs were located at the postsynaptic membrane. The possible implications of these newly described nAChRs in various physiological processes and behavioral events, such as the wake-sleep cycle, are discussed.

  20. Control of breathing by raphe obscurus serotonergic neurons in mice. (United States)

    Depuy, Seth D; Kanbar, Roy; Coates, Melissa B; Stornetta, Ruth L; Guyenet, Patrice G


    We used optogenetics to determine the global respiratory effects produced by selectively stimulating raphe obscurus (RO) serotonergic neurons in anesthetized mice and to test whether these neurons detect changes in the partial pressure of CO(2), and hence function as central respiratory chemoreceptors. Channelrhodopsin-2 (ChR2) was selectively (∼97%) incorporated into ∼50% of RO serotonergic neurons by injecting AAV2 DIO ChR2-mCherry (adeno-associated viral vector double-floxed inverse open reading frame of ChR2-mCherry) into the RO of ePet-Cre mice. The transfected neurons heavily innervated lower brainstem and spinal cord regions involved in autonomic and somatic motor control plus breathing but eschewed sensory related regions. Pulsed laser photostimulation of ChR2-transfected serotonergic neurons increased respiratory frequency (fR) and diaphragmatic EMG (dEMG) amplitude in relation to the duration and frequency of the light pulses (half saturation, 1 ms; 5-10 Hz). dEMG amplitude and fR increased slowly (half saturation after 10-15 s) and relaxed monoexponentially (tau, 13-15 s). The breathing stimulation was reduced ∼55% by methysergide (broad spectrum serotonin antagonist) and potentiated (∼16%) at elevated levels of inspired CO(2) (8%). RO serotonergic neurons, identified by their entrainment to short light pulses (threshold, 0.1-1 ms) were silent (nine cells) or had a low and regular level of activity (2.1 ± 0.4 Hz; 11 cells) that was not synchronized with respiration. These and nine surrounding neurons with similar characteristics were unaffected by adding up to 10% CO(2) to the breathing mixture. In conclusion, RO serotonergic neurons activate breathing frequency and amplitude and potentiate the central respiratory chemoreflex but do not appear to have a central respiratory chemoreceptor function.

  1. Effects of damage to median raphe nucleus on ingestive behavior and wheel running activity. (United States)

    Shahid Salles, M S; Heym, J; Gladfelter, W E


    The effects of damage to the median raphe nucleus on the ingestive behavior and wheel running activity of rats were studied. This nucleus was damaged by the placement of either electrolytic or chemical (5,7-dihydroxytryptamine) lesions. After the placement of either type of lesion, wheel running activity was significantly decreased for the duration of the 8 week post-operative period. Although there were transient decreases in both food and water intakes after damage to the median raphe nucleus, these decreases did not appear to result from impairments in neuro-regulatory mechanisms. Rather, the decrease in food intake seemed to be related to the decrease in locomotor activity, and the decrease in water intake appeared to be linked to the decrease in food intake. In some rats with electrolytic lesions in the median raphe nucleus, the decrease in water intake was followed by a transient period of hyperdipsia.

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

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    Jayesh Sagar


    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.

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

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


    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...

  4. Reward Processing by the Dorsal Raphe Nucleus: 5-HT and Beyond (United States)

    Luo, Minmin; Zhou, Jingfeng; Liu, Zhixiang


    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…

  5. Median raphe cysts in men. Presentation of our experience and literature review. (United States)

    Navalón-Monllor, V; Ordoño-Saiz, M V; Ordoño-Domínguez, F; Sabater-Marco, V; Pallás-Costa, Y; Navalón-Verdejo, P


    To present our experience with the diagnosis and treatment of median raphe cysts treated in our department in the last 25years. We conducted a retrospective study of 28men with median raphe cysts who underwent surgery in our department from June 1990 to March 2015. We analysed the age of presentation, reason for consultation, clinical manifestations, histological findings, treatment and outcome after exeresis. The majority of the patients (22; 79%) were asymptomatic and consulted for the aesthetic defect. Four cases (14%) presented urinary abnormalities, and 2 cases (7%) reported discomfort during sexual intercourse. In all cases, the treatment consisted of surgical extirpation of the cysts, with excellent aesthetic and functional results and no lesion recurrence in any of the patients during a mean follow-up of more than 10years. The most common histological type was the transitional cell type in 15 cases (54%), followed by the mixed type (transitional and squamous) in 11 cases (39%). One case (6%) was pure squamous type, and in another case (6%) the epithelium was glandular. Median raphe cysts are an uncommon type of disembryoplasia that can occur in any location of the median raphe, from the balanic meatus to the edges of the anus. These cysts are generally asymptomatic and their treatment of choice is surgical extirpation. Copyright © 2016 AEU. Publicado por Elsevier España, S.L.U. All rights reserved.

  6. Involvement of the caudal raphe nuclei in the feeding behavior of rats

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    Takase L.F.


    Full Text Available Involvement of the caudal raphe nuclei (raphe pallidus, RPa; raphe magnus, RMg, and raphe obscurus, ROb in feeding behavior of adult rats was studied by measuring c-Fos protein expression, in animals submitted to the "meal-feeding" model of food restriction in which the rats were fed ad libitum only from 7:00 to 9:00 h, for 15 days. The experimental groups submitted to chronic fasting, named 'search for food' (SF, 'ingestion of food' (IF and 'satiety of food' (SaF were scheduled after a previous study in which the body weight and the general and feeding behaviors were evaluated by daily monitoring. Acute, 48-h fasting (AF was used as control. In the chronic group, the animals presented a 16% reduction in body weight in the first week, followed by a continuous, slow rise in weight over the subsequent days. Entrainment of the sleep-wake cycle to the schedule of food presentation was also observed. The RPa was the most Fos immunopositive nucleus in the chronic fasting group, followed by the RMg. The ANOVA and Tukey test (P<0.05 confirmed these results. The IF group was significantly different from the other three groups, as also was the number of labeled cells in the RPa in SF and IF groups. Nevertheless, no significant difference was observed between RMg and RPa, or RMg and ROb in the SaF and AF. However, it is interesting to observe that the groups in which the animals were more active, searching for or ingesting food, presented a larger number of labeled cells. These results suggest a different involvement of the caudal raphe nuclei in the somatic and autonomic events of feeding behavior, corroborating the functions reported for them earlier.

  7. Creating standards: Creating illusions?

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    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...

  8. Functional interactions between KCNE1 C-terminus and the KCNQ1 channel.

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    Jerri Chen

    Full Text Available The KCNE1 gene product (minK protein associates with the cardiac KvLQT1 potassium channel (encoded by KCNQ1 to create the cardiac slowly activating delayed rectifier, I(Ks. Mutations throughout both genes are linked to the hereditary cardiac arrhythmias in the Long QT Syndrome (LQTS. KCNE1 exerts its specific regulation of KCNQ1 activation via interactions between membrane-spanning segments of the two proteins. Less detailed attention has been focused on the role of the KCNE1 C-terminus in regulating channel behavior. We analyzed the effects of an LQT5 point mutation (D76N and the truncation of the entire C-terminus (Delta70 on channel regulation, assembly and interaction. Both mutations significantly shifted voltage dependence of activation in the depolarizing direction and decreased I(Ks current density. They also accelerated rates of channel deactivation but notably, did not affect activation kinetics. Truncation of the C-terminus reduced the apparent affinity of KCNE1 for KCNQ1, resulting in impaired channel formation and presentation of KCNQ1/KCNE1 complexes to the surface. Complete saturation of KCNQ1 channels with KCNE1-Delta70 could be achieved by relative over-expression of the KCNE subunit. Rate-dependent facilitation of K(+ conductance, a key property of I(Ks that enables action potential shortening at higher heart rates, was defective for both KCNE1 C-terminal mutations, and may contribute to the clinical phenotype of arrhythmias triggered by heart rate elevations during exercise in LQTS mutations. These results support several roles for KCNE1 C-terminus interaction with KCNQ1: regulation of channel assembly, open-state destabilization, and kinetics of channel deactivation.

  9. Role of the Oxytocin Receptor Expressed in the Rostral Medullary Raphe in Thermoregulation During Cold Conditions


    Kasahara, Yoshiyuki; Tateishi, Yuko; Hiraoka, Yuichi; Otsuka, Ayano; Mizukami, Hiroaki; Ozawa, Keiya; Sato, Keisuke; Hidema, Shizu; Nishimori, Katsuhiko


    Recent papers have reported that oxytocin (Oxt) and the oxytocin receptor (Oxtr) may be involved in the regulation of food intake in mammals. We therefore suspected the Oxt/Oxtr system to be involved in energy homeostasis. In previous studies, we found a tendency toward obesity in Oxtr-deficient (Oxtr ?/?) mice, as well as impaired thermoregulation when these mice were exposed to cold conditions. In the present study, we observed the expression of Oxtr in the rostral medullary raphe (RMR), th...

  10. Dorsal Raphe Serotonin Neurons Mediate CO2-Induced Arousal from Sleep. (United States)

    Smith, Haleigh R; Leibold, Nicole K; Rappoport, Daniel A; Ginapp, Callie M; Purnell, Benton S; Bode, Nicole M; Alberico, Stephanie L; Kim, Young-Cho; Audero, Enrica; Gross, Cornelius T; Buchanan, Gordon F


    Arousal from sleep in response to CO 2 is a critical protective phenomenon. Dysregulation of CO 2 -induced arousal contributes to morbidity and mortality from prevalent diseases, such as obstructive sleep apnea and sudden infant death syndrome. Despite the critical nature of this protective reflex, the precise mechanism for CO 2 -induced arousal is unknown. Because CO 2 is a major regulator of breathing, prevailing theories suggest that activation of respiratory chemo- and mechano-sensors is required for CO 2 -induced arousal. However, populations of neurons that are not involved in the regulation of breathing are also chemosensitive. Among these are serotonin (5-HT) neurons in the dorsal raphe nucleus (DRN) that comprise a component of the ascending arousal system. We hypothesized that direct stimulation of these neurons with CO 2 could cause arousal from sleep independently of enhancing breathing. Dialysis of CO 2 -rich acidified solution into DRN, but not medullary raphe responsible for modulating breathing, caused arousal from sleep. Arousal was lost in mice with a genetic absence of 5-HT neurons, and with acute pharmacological or optogenetic inactivation of DRN 5-HT neurons. Here we demonstrate that CO 2 can cause arousal from sleep directly, without requiring enhancement of breathing, and that chemosensitive 5-HT neurons in the DRN critically mediate this arousal. Better understanding mechanisms underlying this protective reflex may lead to interventions to reduce disease-associated morbidity and mortality. SIGNIFICANCE STATEMENT Although CO 2 -induced arousal is critical to a number of diseases, the specific mechanism is not well understood. We previously demonstrated that serotonin (5-HT) neurons are important for CO 2 -induced arousal, as mice without 5-HT neurons do not arouse to CO 2 Many have interpreted this to mean that medullary 5-HT neurons that regulate breathing are important in this arousal mechanism. Here we found that direct application of CO 2

  11. 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


    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

  12. Deviation of the penoscrotal median raphe: Is it a normal finding or within the spectrum of hypospadias?

    Directory of Open Access Journals (Sweden)

    Arvind Mohan


    Full Text Available Introduction and Objectives: Hypospadias is the most common congenital abnormality of the penis, and is most commonly diagnosed during the postnatal physical examination. However, milder forms of the condition can be difficult to detect, leading to delayed referral to specialist teams. We aim to determine whether there is an association between hypospadias and the position of the penoscrotal raphe. Materials and Methods: A case - control study was performed where clinical photographs from children undergoing hypospadias correction were compared with a control group of children without the condition. The position of the penoscrotal raphe was documented as midline, left or right. Pearson′s chi squared test was used to determine significance. Results: Images for 80 children undergoing hypospadias correction were compared with 80 normal children in the maternity ward. 88.8% of the children with hypospadias had a penoscrotal raphe deviated from the midline compared with only 13.8% in the control group (P < 0.0003. Conclusions: Our study demonstrates a significant association between hypospadias and deviation of the penoscrotal raphe from the midline. Consideration should be given to whether to include this finding within the spectrum of abnormalities seen in hypospadias. Examination of the penoscrotal raphe is simple to perform and could aid in the early diagnosis in children with milder forms of the condition.

  13. Deficiency of Serotonin in Raphe Neurons and Altered Behavioral Responses in Tryptophan Hydroxylase 2-Knockout Medaka (Oryzias latipes). (United States)

    Ansai, Satoshi; Hosokawa, Hiroshi; Maegawa, Shingo; Naruse, Kiyoshi; Washio, Youhei; Sato, Kenji; Kinoshita, Masato


    Serotonin (5-hydroxytryptamine [5-HT]) is a bioactive monoamine that acts as a neurotransmitter in the central and peripheral nervous system of animals. Teleost fish species have serotonergic neurons in the raphe nuclei of the brainstem; however, the role of 5-HT in the raphe neurons in teleost fish remains largely unknown. Here, we established a medaka (Oryzias latipes) strain with targeted disruption of tryptophan hydroxylase 2 (tph2) gene that is involved in the 5-HT synthesis in the raphe nuclei. Immunohistochemistry and mass spectrometry analysis revealed that the homozygous mutants (tph2Δ13/Δ13) lacked the ability to synthesize 5-HT in the raphe neurons. To investigate the effects of 5-HT deficiency in adult behaviors, the mutant fish were subjected to five behavioral paradigms (diving, open-field, light-dark transition, mirror-biting, and two-fish social interaction). The homozygous mutation caused a longer duration of freezing response in all examined paradigms and reduced the number of entries to the top area in the diving test. In addition, the mutants exhibited a decreased number of mirror-biting in the males and an increased contact time in direct social interaction between the females. These results indicate that this tph2-knockout medaka serves as a good model to analyze the effects of 5-HT deficiency in the raphe neurons.

  14. Activation of dorsal raphe serotonin neurons is necessary for waiting for delayed rewards. (United States)

    Miyazaki, Kayoko W; Miyazaki, Katsuhiko; Doya, Kenji


    The forebrain serotonergic system is a crucial component in the control of impulsive behaviors. We previously reported that the activity of serotonin neurons in the midbrain dorsal raphe nucleus increased when rats performed a task that required them to wait for delayed rewards. However, the causal relationship between serotonin neural activity and the tolerance for the delayed reward remained unclear. Here, we test whether the inhibition of serotonin neural activity by the local application of the 5-HT(1A) receptor agonist 8-hydroxy-2-(di-n-propylamino) tetralin in the dorsal raphe nucleus impairs rats' tolerance for delayed rewards. Rats performed a sequential food-water navigation task that required them to visit food and water sites alternately via a tone site to get rewards at both sites after delays. During the short (2 s) delayed reward condition, the inhibition of serotonin neural activity did not significantly influence the numbers of reward choice errors (nosepoke at an incorrect reward site following a conditioned reinforcer tone), reward wait errors (failure to wait for the delayed rewards), or total trials (sum of reward choice errors, reward wait errors, and acquired rewards). By contrast, during the long (7-11 s) delayed reward condition, the number of wait errors significantly increased while the numbers of total trials and choice errors did not significantly change. These results indicate that the activation of dorsal raphe serotonin neurons is necessary for waiting for long delayed rewards and suggest that elevated serotonin activity facilitates waiting behavior when there is the prospect of forthcoming rewards.

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

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    Masoumeh Kourosh Arami


    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.

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

    DEFF Research Database (Denmark)

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


    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......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...

  17. Respiratory and Mayer wave-related discharge patterns of raphé and pontine neurons change with vagotomy. (United States)

    Morris, K F; Nuding, S C; Segers, L S; Baekey, D M; Shannon, R; Lindsey, B G; Dick, T E


    Previous models have attributed changes in respiratory modulation of pontine neurons after vagotomy to a loss of pulmonary stretch receptor "gating" of an efference copy of inspiratory drive. Recently, our group confirmed that pontine neurons change firing patterns and become more respiratory modulated after vagotomy, although average peak and mean firing rates of the sample did not increase (Dick et al., J Physiol 586: 4265-4282, 2008). Because raphé neurons are also elements of the brain stem respiratory network, we tested the hypotheses that after vagotomy raphé neurons have increased respiratory modulation and that alterations in their firing patterns are similar to those seen for pontine neurons during withheld lung inflation. Raphé and pontine neurons were recorded simultaneously before and after vagotomy in decerebrated cats. Before vagotomy, 14% of 95 raphé neurons had increased activity during single respiratory cycles prolonged by withholding lung inflation; 13% exhibited decreased activity. After vagotomy, the average index of respiratory modulation (eta(2)) increased (0.05 +/- 0.10 to 0.12 +/- 0.18 SD; Student's paired t-test, P waves. These "Mayer wave-related oscillations" (MWROs) were coupled with central respiratory drive and became synchronized with the central respiratory rhythm after vagotomy (7 of 10 animals). Cross-correlation analysis identified functional connectivity in 52 of 360 pairs of neurons with MWROs. Collectively, the results suggest that a distributed network participates in the generation of MWROs and in the coordination of respiratory and vasomotor rhythms.

  18. Prenatal stress alters diazepam withdrawal syndrome and 5HT1A receptor expression in the raphe nuclei of adult rats. (United States)

    Lakehayli, S; Said, N; El Khachibi, M; El Ouahli, M; Nadifi, S; Hakkou, F; Tazi, A


    Early-life events have long-term effects on brain structures and cause behavioral alterations that persist into adulthood. The present experiments were designed to investigate the effects of prenatal stress on diazepam-induced withdrawal syndrome and serotonin-1A (5HT1A) receptor expression in the raphe nuclei of adult offspring. The results of the present study reveal that maternal exposure to chronic footshock stress increased the anxiety-like behavior in the prenatally stressed (PS) animals withdrawn from chronic diazepam (2.5mg/kg/day i.p for 1week). Moreover, prenatal stress induced a down-regulation of 5HT1A mRNA in the raphe nuclei of adult offspring. To our knowledge, this study is the first to demonstrate that maternal exposure to chronic footshock stress enhances diazepam withdrawal symptoms and alters 5HT1A receptor gene expression in the raphe nuclei of adult offspring. Thus, more studies are needed to clarify the mechanisms underlying the decrease of 5HT1A receptors expression in the raphe nuclei of PS rats. Copyright © 2016 IBRO. Published by Elsevier Ltd. All rights reserved.

  19. Regulation of the release of serotonin in the dorsal raphe nucleus by alpha(1) and alpha(2) adrenoceptors

    NARCIS (Netherlands)

    Pudovkina, OL; Cremers, TIFH; Westerink, BHC


    To investigate the modulation of serotonin release in the dorsal raphe nucleus (DRN) by alpha(1) and alpha(2) adrenoceptors, dual-probe microdialysis was performed in conscious rats. The specific alpha(1) and alpha(2) adrenoceptor agonists and antagonists were locally infused into the DRN via

  20. 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


    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...

  1. Cell-Type-Specific Modulation of Sensory Responses in Olfactory Bulb Circuits by Serotonergic Projections from the Raphe Nuclei. (United States)

    Brunert, Daniela; Tsuno, Yusuke; Rothermel, Markus; Shipley, Michael T; Wachowiak, Matt


    Serotonergic neurons in the brainstem raphe nuclei densely innervate the olfactory bulb (OB), where they can modulate the initial representation and processing of olfactory information. Serotonergic modulation of sensory responses among defined OB cell types is poorly characterized in vivo Here, we used cell-type-specific expression of optical reporters to visualize how raphe stimulation alters sensory responses in two classes of GABAergic neurons of the mouse OB glomerular layer, periglomerular (PG) and short axon (SA) cells, as well as mitral/tufted (MT) cells carrying OB output to piriform cortex. In PG and SA cells, brief (1-4 s) raphe stimulation elicited a large increase in the magnitude of responses linked to inhalation of ambient air, as well as modest increases in the magnitude of odorant-evoked responses. Near-identical effects were observed when the optical reporter of glutamatergic transmission iGluSnFR was expressed in PG and SA cells, suggesting enhanced excitatory input to these neurons. In contrast, in MT cells imaged from the dorsal OB, raphe stimulation elicited a strong increase in resting GCaMP fluorescence with only a slight enhancement of inhalation-linked responses to odorant. Finally, optogenetically stimulating raphe serotonergic afferents in the OB had heterogeneous effects on presumptive MT cells recorded extracellularly, with an overall modest increase in resting and odorant-evoked responses during serotonergic afferent stimulation. These results suggest that serotonergic afferents from raphe dynamically modulate olfactory processing through distinct effects on multiple OB targets, and may alter the degree to which OB output is shaped by inhibition during behavior. Modulation of the circuits that process sensory information can profoundly impact how information about the external world is represented and perceived. This study investigates how the serotonergic system modulates the initial processing of olfactory information by the

  2. More tryptophan hydroxylase in the brainstem dorsal raphe nucleus in depressed suicides. (United States)

    Boldrini, Maura; Underwood, Mark D; Mann, J John; Arango, Victoria


    Deficient serotonin neurotransmission in suicide is indicated by reduced brainstem serotonin (5-HT) and 5-hydroxyindoleacetic acid (5-HIAA), fewer 5-HT(1A) autoreceptors and reduced cortical serotonin transporter binding in suicide victims. Tryptophan hydroxylase (TPH) is the rate-limiting enzyme in the synthesis of 5-HT, and alterations in TPH could explain some of these findings. We sought to determine the amount of TPH immunoreactivity (TPH-IR) in the dorsal (DRN) and median (MRN) raphe nuclei in suicides and controls. Brainstems of suicide victims and controls (n = 11 pairs) were collected at autopsy, matched for age, sex and postmortem interval, frozen and sectioned (20 microm). Immunoautoradiography, using an antibody to label TPH, was performed, slides exposed to film and autoradiograms quantified by a computer-based image analysis system. We examined sections every 1000 microm throughout the whole length of the nucleus, performing statistical analysis only on those subjects for whom the raphe was complete (n = 8 pairs). TPH-IR (microCi/g) was higher in suicides than controls (S: 300.8 +/- 70.8 vs. C: 259.6 +/- 40.7, t = 2.57, df = 7, P = 0.04) in the dorsal raphe nucleus (DRN), and not different between suicides and controls (S: 251.3 +/- 44.2 vs. C: 235.9 +/- 27.4, t = 1.49, df = 7, P = 0.18) in the MRN. DRN TPH-IR was higher in male suicide victims (MS) compared to male controls (MC; MS: 318.4 +/- 54.4 vs. MC: 271.9 +/- 22.5, t = 2.66, df = 6, P = 0.03). The analysis of TPH-IR area and density at each DRN rostrocaudal levels showed higher area and density in suicides compared to controls in the rostral DRN and lower area and density in the caudal DRN. TPH-IR, an index of the amount of TPH enzyme, in the DRN is higher in depressed suicides. More TPH may be an upregulatory homeostatic response to impaired serotonin release or less autoreceptor activation. Alternatively, the serotonin impairment in suicide may be due to hypofunctional serotonin

  3. Raphe serotonin neuron-specific oxytocin receptor knockout reduces aggression without affecting anxiety-like behavior in male mice only. (United States)

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


    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.

  4. Intense Activity of the Raphe Spinal Pathway Depresses Motor Activity via a Serotonin Dependent Mechanism

    DEFF Research Database (Denmark)

    Perrier, Jean-François; Rasmussen, Hanne B; Jørgensen, Lone K


    Motor fatigue occurring during prolonged physical activity has both peripheral and central origins. It was previously demonstrated that the excitability of motoneurons was decreased when a spillover of serotonin could activate extrasynaptic 5-HT1A receptors at the axon initial segment (AIS......) of motoneurons. Here we investigated the impact of massive synaptic release of serotonin on motor behavior in an integrated preparation of the adult turtle performing fictive scratching behaviors. We found that a prolonged electrical stimulation of the raphe spinal pathway induced a reversible inhibition...... of the motor behavior that lasted several tens of seconds. The effect disappeared when the spinal cord was perfused with an antagonist for 5-HT1A receptors. By demonstrating a direct impact of serotonin on motor behavior, we suggest a central role of this monoamine behind central fatigue....

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

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    Toshikazu Ikuta


    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.

  6. 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


    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.

  7. Creating Poetry. (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.…

  8. LHC Create

    CERN Multimedia

    CERN. Geneva


    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.

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

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


    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.

  10. Central and peripheral chemoreceptors evoke distinct responses in simultaneously recorded neurons of the raphé-pontomedullary respiratory network. (United States)

    Nuding, Sarah C; Segers, Lauren S; Shannon, Roger; O'Connor, Russell; Morris, Kendall F; Lindsey, Bruce G


    The brainstem network for generating and modulating the respiratory motor pattern includes neurons of the medullary ventrolateral respiratory column (VRC), dorsolateral pons (PRG) and raphé nuclei. Midline raphé neurons are proposed to be elements of a distributed brainstem system of central chemoreceptors, as well as modulators of central chemoreceptors at other sites, including the retrotrapezoid nucleus. Stimulation of the raphé system or peripheral chemoreceptors can induce a long-term facilitation of phrenic nerve activity; central chemoreceptor stimulation does not. The network mechanisms through which each class of chemoreceptor differentially influences breathing are poorly understood. Microelectrode arrays were used to monitor sets of spike trains from 114 PRG, 198 VRC and 166 midline neurons in six decerebrate vagotomized cats; 356 were recorded during sequential stimulation of both receptor classes via brief CO(2)-saturated saline injections in vertebral (central) and carotid arteries (peripheral). Seventy neurons responded to both stimuli. More neurons were responsive only to peripheral challenges than those responsive only to central chemoreceptor stimulation (PRG, 20 : 4; VRC, 41 : 10; midline, 25 : 13). Of 16 474 pairs of neurons evaluated for short-time scale correlations, similar percentages of reference neurons in each brain region had correlation features indicative of a specific interaction with at least one target neuron: PRG (59.6%), VRC (51.0%) and raphé nuclei (45.8%). The results suggest a brainstem network architecture with connectivity that shapes the respiratory motor pattern via overlapping circuits that modulate central and peripheral chemoreceptor-mediated influences on breathing.

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


    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


    Background 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). Methods Rats were given acupuncture at ST41 (Jiexi), LI11 (Quchi) or SI3 (Houxi) acupoint in LPS-treated ...

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

    Directory of Open Access Journals (Sweden)

    A Sanchez-Bahillo


    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

  13. Distinct Contributions of Median Raphe Nucleus to Contextual Fear Conditioning and Fear-Potentiated Startle (United States)

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


    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

  14. Anxiolytic-like effect of mirtazapine mediates its effect in the median raphe nucleus. (United States)

    An, Yan; Inoue, Takeshi; Kitaichi, Yuji; Izumi, Takeshi; Nakagawa, Shin; Song, Ning; Chen, Chong; Li, XiaoBai; Koyama, Tsukasa; Kusumi, Ichiro


    Mirtazapine, a noradrenergic and specific serotonergic antidepressant (NaSSA), blocks the α2-adrenergic autoreceptors and heteroreceptors, which are responsible for controlling noradrenaline and 5-hydroxy-tryptamine (5-HT) release. Though preclinical and clinical studies have shown that mirtazapine exerts an anxiolytic action, its precise brain target sites remain unclear. In the present study, we investigated the brain area(s) in which mirtazapine exerts its anxiolytic-like effects on the expression of contextual conditioned freezing in rats. Mirtazapine (3 μg/site) was directly injected into three brain structures, the median raphe nucleus (MRN), hippocampus and amygdala. Freezing behavior tests were carried out 10 min after injections. Our results showed that the intra-MRN injection of mirtazapine reduced freezing significantly, whereas injections into the hippocampus or the amygdala did not. In addition, the intra-MRN injection of mirtazapine did not affect locomotor activity. These results suggest that the anxiolytic-like effect of mirtazapine might be mediated by its action on the MRN.

  15. A pilot study on predictors of brainstem raphe abnormality in patients with major depressive disorder. (United States)

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


    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.

  16. Extrasynaptic glycine receptors of rodent dorsal raphe serotonergic neurons: a sensitive target for ethanol. (United States)

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


    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 inhibitory 'phasic' post-synaptic currents mediated primarily by synaptic GABAA receptors (GABAAR) and, to a lesser extent, by synaptic glycine receptors (GlyR). In addition to such phasic transmission mediated by the vesicular release of neurotransmitter, the activity of certain neurons may be governed by a 'tonic' conductance resulting from ambient GABA activating extrasynaptic GABAARs. However, for DR neurons extrasynaptic GABAARs exert only a limited influence. By contrast, we report that unusually the GlyR antagonist strychnine reveals a large tonic conductance mediated by extrasynaptic GlyRs, which dominates DR inhibition. In agreement, for DR neurons strychnine increases their input resistance, induces membrane depolarization, and consequently augments their excitability. Importantly, this glycinergic conductance is greatly enhanced in a strychnine-sensitive fashion, by behaviorally relevant ethanol concentrations, by drugs used for the treatment of alcohol withdrawal, and by taurine, an ingredient of certain 'energy drinks' often imbibed with ethanol. These findings identify extrasynaptic GlyRs as critical regulators of DR excitability and a novel molecular target for ethanol.

  17. Increased mRNA expression of cytochrome oxidase in dorsal raphe nucleus of depressive suicide victims (United States)

    Sanchez-Bahillo, A; Bautista-Hernandez, V; Barcia Gonzalez, Carlos; Bañon, R; Luna, A; Hirsch, EC; Herrero, Maria-Trinidad


    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 serotonin re-uptake. No alteration was found in noradrenergic neurons, suggesting that they play no crucial role in the suicidal behavior of depressive patients. PMID:18728743

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

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    Merab G Tsagareli


    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.

  19. Corticotropin-releasing factor in the dorsal raphe nucleus: Linking stress coping and addiction. (United States)

    Valentino, Rita J; Lucki, Irwin; Van Bockstaele, Elisabeth


    Addiction and stress are linked at multiple levels. Drug abuse is often initiated as a maladaptive mechanism for coping with stress. It is maintained in part by negative reinforcement to prevent the aversive consequences of stress associated with abstinence. Finally, stress is a major factor leading to relapse in subjects in which drug seeking behavior has extinguished. These associations imply overlapping or converging neural circuits and substrates that underlie the processes of addiction and the expression of the stress response. Here we discuss the major brain serotonin (5-HT) system, the dorsal raphe nucleus (DRN)-5-HT system as a point of convergence that links these processes and how the stress-related neuropeptide, corticotropin-releasing factor (CRF) directs this by a bimodal regulation of DRN neuronal activity. The review begins by describing a structural basis for CRF regulation of the DRN-5-HT system. This is followed by a review of the effects of CRF and stress on DRN function based on electrophysiological and microdialysis studies. The concept that multiple CRF receptor subtypes in the DRN facilitate distinct coping behaviors is reviewed with recent evidence for a unique cellular mechanism by which stress history can determine the type of coping behavior. Finally, work on CRF regulation of the DRN-5-HT system is integrated with literature on the role of 5-HT-dopamine interactions in addiction. Copyright 2009 Elsevier B.V. All rights reserved.

  20. Normal levels of tryptophan hydroxylase immunoreactivity in the dorsal raphe of depressed suicide victims. (United States)

    Bonkale, Willy L; Murdock, Shayna; Janosky, Janine E; Austin, Mark C


    A variety of evidence suggests that serotonin neurotransmission is altered in the brain of suicide victims and depressed patients. While numerous post-mortem studies have investigated serotonin transporters and receptors, few studies have examined the biosynthetic integrity of the rate-limiting enzyme, tryptophan hydroxylase (TPH), in post-mortem specimens of depressed suicide subjects. Therefore, the aim of the present study was to test the hypothesis that the levels of TPH immunoreactivity (IR) are altered in specific subnuclei of the dorsal raphe (DR) in depressed suicide victims. Suicide victims with a confirmed diagnosis of major depression were matched with non-psychiatric controls based on age, gender and post-mortem interval. Frozen tissue sections containing the DR were selected from two anatomical levels and processed for TPH radioimmunocytochemistry. The optical density corresponding to the regional levels of TPH-IR was quantified in specific subnuclei of the DR from the film autoradiographic images. No significant differences in the levels of TPH-IR were found in any DR subnuclei between depressed suicide victims and control subjects. The lack of change in TPH-IR levels does not necessarily imply that serotonin synthesis or neurotransmission is not altered in the brain of depressed subjects. Many factors influence and regulate serotonin synthesis, and it is conceivable that alterations exist at other levels of regulation of serotonin biosynthesis in depression. Our findings indicate that TPH biosynthesis, at least at the protein level, is not significantly altered in the DR of depressed suicide victims.

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

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    Ross A. McDevitt


    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.

  2. Biophysical properties and computational modeling of calcium spikes in serotonergic neurons of the dorsal raphe nucleus. (United States)

    Tuckwell, Henry C


    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.

  3. Role of the oxytocin receptor expressed in the rostral medullary raphe in thermoregulation during cold conditions

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    Yoshiyuki eKasahara


    Full Text Available Recent papers have reported that oxytocin (Oxt and the oxytocin receptor (Oxtr may be involved in the regulation of food intake in mammals. We therefore suspected the Oxt/Oxtr system to be involved in energy homeostasis. In previous studies, we found a tendency toward obesity in Oxtr-deficient mice, as well as impaired thermoregulation when these mice were exposed to cold conditions. In the present study, we observed the expression of Oxtr in the rostral medullary raphe (RMR, the brain region known to control thermogenesis in brown adipose tissue. Through immunohistochemistry, we detected neurons expressing Oxtr and c-Fos in the RMR of mice exposed to cold conditions. Up to 40% of Oxtr-positive neurons in RMR were classified as glutamatergic neurons, as shown by immunostaining using anti-VGLUT3 antibody. In addition, mice with exclusive expression of Oxtr in the RMR were generated by injecting an AAV-Oxtr vector into the RMR region of Oxtr-deficient mice. We confirmed the recovery of thermoregulatory ability in the manipulated mice during exposure to cold conditions. Moreover, mice with RMR-specific expression of Oxtr lost the typical morphological change in brown adipose tissue observed in Oxtr-deficient mice. Additionally, increased expression of the β3-adrenergic receptor gene, Adrb3 was observed in brown adipose tissue. These results are the first to show the critical role of RMR Oxtr expression in thermoregulation during cold conditions.

  4. Different actions for acute and chronic administration of mirtazapine on serotonergic transmission associated with raphe nuclei and their innervation cortical regions. (United States)

    Yamamura, Satoshi; Abe, Masao; Nakagawa, Masanori; Ochi, Shinichiro; Ueno, Shu-ichi; Okada, Motohiro


    The atypical antidepressant, mirtazapine enhances noradrenergic transmission, but its effects on serotonergic transmission remain to be clarified. The present study determined the effects of acute and chronic administration of mirtazapine on serotonergic transmissions in raphe nuclei and their innervation regions, frontal and entorhinal cortex, using multiple-probes microdialysis with real-time PCR and western blotting. Acute administration of mirtazapine did not affect extracellular serotonin level in raphe nuclei or cortex; however, chronic administration increased extracellular serotonin level in raphe nuclei without affecting that in cortex. Blockade of 5-HT1A receptor, but not that of the 5-HT2A/2C receptor, enhanced the effects of acute administration of mirtazapine on extracellular serotonin level in raphe nuclei. Chronic mirtazapine administration reduced the inhibitory function associated with somatodendritic 5-HT1A receptor in raphe nuclei, but enhanced postsynaptic 5-HT1A receptor in serotonergic innervated cortical regions. Chronic administration reduced the expression of mRNA and protein of serotonin transporter and 5-HT1A receptor in raphe nuclei, but not in the cortices. These results suggested that acute administration of mirtazapine probably activated serotonergic transmission, but its stimulatory action was abolished by activated inhibitory 5-HT1A receptor. Chronic administration of mirtazapine resulted in increased extracellular serotonin level via reduction of serotonin transporter with reduction of somatodendritic 5-HT1A autoreceptor function in raphe nuclei. These pharmacological actions of mirtazapine include its serotonergic profiles as noradrenergic and specific serotonergic antidepressant (NaSSA). Copyright © 2011 Elsevier Ltd. All rights reserved.

  5. Francis Bacon's Valerius Terminus and the Voyage to the "Great Instauration". (United States)

    Serjeantson, Richard


    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.

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


    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.

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

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    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.

  8. Nicotine increases GABAergic input on rat dorsal raphe serotonergic neurons through alpha7 nicotinic acetylcholine receptor. (United States)

    Hernández-Vázquez, F; Chavarría, K; Garduño, J; Hernández-López, S; Mihailescu, S P


    The dorsal raphe nucleus (DRN) contains large populations of serotonergic (5-HT) neurons. This nucleus receives GABAergic inhibitory afferents from many brain areas and from DRN interneurons. Both GABAergic and 5-HT DRN neurons express functional nicotinic acetylcholine receptors (nAChRs). Previous studies have demonstrated that nicotine increases 5-HT release and 5-HT DRN neuron discharge rate by stimulating postsynaptic nAChRs and by increasing glutamate and norepinephrine release inside DRN. However, the influence of nicotine on the GABAergic input to 5-HT DRN neurons was poorly investigated. Therefore, the aim of this work was to determine the effect of nicotine on GABAergic spontaneous inhibitory postsynaptic currents (sIPSCs) of 5-HT DRN neurons and the subtype of nAChR(s) involved in this response. Experiments were performed in coronal slices obtained from young Wistar rats. GABAergic sIPSCs were recorded from post hoc-identified 5-HT DRN neurons with the whole cell voltage patch-clamp technique. Administration of nicotine (1 μM) increased sIPSC frequency in 72% of identified 5-HT DRN neurons. This effect was not reproduced by the α4β2 nAChR agonist RJR-2403 and was not influenced by TTX (1 μM). It was mimicked by the selective agonist for α7 nAChR, PNU-282987, and exacerbated by the positive allosteric modulator of the same receptor, PNU-120596. The nicotine-induced increase in sIPSC frequency was independent on voltage-gated calcium channels and dependent on Ca(2+)-induced Ca(2+) release (CICR). These results demonstrate that nicotine increases the GABAergic input to most 5-HT DRN neurons, by activating α7 nAChRs and producing CICR in DRN GABAergic terminals. Copyright © 2014 the American Physiological Society.

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

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    Kae eNakamura


    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.

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

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    Cavalcante-Lima H.R.


    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.

  11. The Alteration of Neonatal Raphe Neurons by Prenatal-Perinatal Nicotine. Meaning for Sudden Infant Death Syndrome. (United States)

    Cerpa, Verónica J; Aylwin, María de la Luz O; Beltrán-Castillo, Sebastián; Bravo, Eduardo U; Llona, Isabel R; Richerson, George B; Eugenín, Jaime L


    Nicotine may link maternal cigarette smoking with respiratory dysfunctions in sudden infant death syndrome (SIDS). Prenatal-perinatal nicotine exposure blunts ventilatory responses to hypercapnia and reduces central respiratory chemoreception in mouse neonates at Postnatal Days 0 (P0) to P3. This suggests that raphe neurons, which are altered in SIDS and contribute to central respiratory chemoreception, may be affected by nicotine. We therefore investigated whether prenatal-perinatal nicotine exposure affects the activity, electrical properties, and chemosensitivity of raphe obscurus (ROb) neurons in mouse neonates. Osmotic minipumps, implanted subcutaneously in 5- to 7-day-pregnant CF1 mice, delivered nicotine bitartrate (60 mg kg(-1) d(-1)) or saline (control) for up to 28 days. In neonates, ventilation was recorded by head-out plethysmography, c-Fos (neuronal activity marker), or serotonin autoreceptors (5HT1AR) were immunodetected using light microscopy, and patch-clamp recordings were made from raphe neurons in brainstem slices under normocarbia and hypercarbia. Prenatal-perinatal nicotine exposure decreased the hypercarbia-induced ventilatory responses at P1-P5, reduced both the number of c-Fos-positive ROb neurons during eucapnic normoxia at P1-P3 and their hypercapnia-induced recruitment at P3, increased 5HT1AR immunolabeling of ROb neurons at P3-P5, and reduced the spontaneous firing frequency of ROb neurons at P3 without affecting their CO2 sensitivity or their passive and active electrical properties. These findings reveal that prenatal-perinatal nicotine reduces the activity of neonatal ROb neurons, likely as a consequence of increased expression of 5HT1ARs. This hypoactivity may change the functional state of the respiratory neural network leading to breathing vulnerability and chemosensory failure as seen in SIDS.

  12. Molecular cloning and expression of the C-terminus of spider ...

    Indian Academy of Sciences (India)

    A cDNA coding for the C-terminus of spider flagelliform silk protein (AvFlag) was cloned from Araneus ventricosus. Analysis of the cDNA sequence shows that the C-terminus of AvFlag consists of 167 amino acids of a repetitive region and 87 amino acids of a C-terminal non-repetitive region. The peptide motifs found in ...

  13. Snca and Bdnf gene expression in the VTA and raphe nuclei of midbrain in chronically victorious and defeated male mice.

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    Natalia N Kudryavtseva

    Full Text Available BACKGROUND: Alpha-synuclein (α-Syn is a small neuronal protein that has been found to be expressed throughout the brain. It has been shown that α-Syn regulates the homeostasis of monoamine neurotransmitters and is involved in various degenerative and affective disorders. There is indication that α-Syn may regulate expression of the brain-derived neurotropic factor (BDNF which plays an important role in the mood disorders. METHODOLOGY/PRINCIPAL FINDINGS: The study aimed to analyze the mRNA levels of Snca and Bdnf genes in the ventral tegmental area (VTA and raphe nuclei of the midbrain in male mice that had each won or defeated 20 encounters (20-time winners and 20-time losers, respectively in daily agonistic interactions. Groups of animals that had the same winning and losing track record followed by a no-fight period for 14 days (no-fighting winners and no-fighting losers were also studied. Snca mRNA levels were increased in the raphe nuclei in the 20-time losers and in the VTA of the 20-time winners. After no-fight period Snca mRNA levels decreased in both groups. Snca mRNA levels were similar to the control level in the VTA of the 20-time losers and in the raphe nuclei of the 20-time winners. However Snca gene expression increased in these areas in the no-fighting winners and no-fighting losers in comparison with respective mRNA levels in animals before no-fight period. Bdnf mRNA levels increased in VTA of 20-time winners. Significant positive correlations were found between the mRNA levels of Snca and Bdnf genes in the raphe nuclei. CONCLUSIONS/SIGNIFICANCE: Social experience affects Snca gene expression depending on brain areas and functional activity of monoaminergic systems in chronically victorious or defeated mice. These findings may be useful for understanding the mechanisms of forming different alpha-synucleinopathies.

  14. Short-term cold exposure activates TRH neurons exclusively in the hypothalamic paraventricular nucleus and raphe pallidus. (United States)

    Cabral, Agustina; Valdivia, Spring; Reynaldo, Mirta; Cyr, Nicole E; Nillni, Eduardo A; Perello, Mario


    The neuropeptide thyrotropin releasing hormone (TRH) is necessary for adequate cold-induced thermogenesis. TRH increases body temperature via both neuroendocrine and autonomic mechanisms. TRH neurons of the hypothalamic paraventricular nucleus (PVN) regulate thermogenesis through the activation of the hypothalamic-pituitary-thyroid axis during cold exposure. However, little is known about the role that TRH neurons play in mediating the sympathetic response to cold exposure. Here, we examined the response of TRH neurons of rats to cold exposure in hypothalamic regions including the PVN, the dorsomedial nucleus and the lateral hypothalamus along with areas of the ventral medulla including raphe obscurus, raphe pallidus (RPa) and parapyramidal regions. Our results using a double immunohistochemistry protocol to identify TRH and c-Fos (as a marker of cellular activity) followed by analysis of preproTRH gene expression demonstrate that only TRH neurons located in the PVN and the RPa are activated in animals exposed to short-term cold conditions. Copyright © 2012 Elsevier Ireland Ltd. All rights reserved.

  15. Evidence for involvement of the subcoeruleus nucleus and nucleus raphe magnus in urine storage and penile erection in decerebrate rats. (United States)

    Sugaya, K; Ogawa, Y; Hatano, T; Koyama, Y; Miyazato, T; Oda, M


    Micturition and male sexual activity require the lower urinary tract to function. During the sexual act, micturition must be inhibited and urine stored in the bladder. We studied the role of the brainstem in relation to both micturition/urine storage and penile erection in rats. Wire electrodes were placed on the dorsal nerve of the penis and microelectrodes for stimulation were introduced into the brainstem in decerebrate male rats. Electrical stimulation was used to locate optimally responding sites by monitoring the isovolumetric intravesical pressure and intracavernous pressure. Electrical stimulation of the dorsal nerve of the penis, the subcoeruleus nucleus in the rostral pons, and the nucleus raphe magnus in the caudal pons increased intracavernous pressure, but inhibited rhythmic bladder contractions. Electrical stimulation of Barrington's nucleus (the pontine micturition center in the rat) in the rostral pons induced bladder contraction. Stimulation of the pontine reticular formation did not increase intracavernous pressure. Acute transection of the thoracic spinal cord eliminated rhythmic bladder contractions, but gave rise to sporadic increments of intracavernous pressure. This electrophysiological study demonstrated that the subcoeruleus nucleus and nucleus raphe magnus are involved in both urine storage and penile erection, and that their physiological functions are reciprocally controlled; so that erection leads to inhibition of micturition.

  16. Motherhood and infant contact regulate neuroplasticity in the serotonergic midbrain dorsal raphe. (United States)

    Holschbach, M Allie; Lonstein, Joseph S


    The adult brain shows remarkable neuroplasticity in response to hormones and the socioemotional modifications that they influence. In females with reproductive and maternal experience, this neuroplasticity includes the birth and death of cells in several forebrain regions involved in maternal caregiving and postpartum affective state. Such plasticity in midbrain sites critical for these behavioral and emotional processes has never been examined, though. By visualizing bromodeoxyuridine (BrdU) to label mitotic cells, NeuroD for neuronal precursors, and TUNEL to identify dying cells, we found that the midbrain dorsal raphe nucleus (DR, the source of most ascending serotoninergic projections) exhibited significant neuroplasticity in response to motherhood. Specifically, BrdU analyses revealed that DR newborn cell survival (but not proliferation) was regulated by reproductive state, such that cells born early postpartum were less likely to survive 12 days to reach the late postpartum period compared to cells born during late pregnancy that survived 12 days to reach the early postpartum period. Many of the surviving cells in the DR were NeuN immunoreactive, suggesting a neuronal phenotype. Consistent with these findings, late postpartum rats had fewer NeuroD-immunoreactive DR cells than early postpartum rats. Maternal experience contributed to the late postpartum reduction in DR newborn cell survival because removing the litter at parturition increased cell survival as well as reduced cell death. Unlike cytogenesis in the maternal hippocampus, which is reduced by circulating glucocorticoids, DR newborn cell survival was unaffected by postpartum adrenalectomy. These effects of reproductive state and motherhood on DR plasticity were associated with concurrent changes in DR levels of serotonin's precursor, 5-HTP, and its metabolite, 5-HIAA. Our results demonstrate for the first time that cytogenesis occurs in the midbrain DR of any adult mammal, that DR plasticity is

  17. 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


    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.

  18. 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.

  19. 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

  20. Computational modeling of spike generation in serotonergic neurons of the dorsal raphe nucleus. (United States)

    Tuckwell, Henry C; Penington, Nicholas J


    Serotonergic neurons of the dorsal raphe nucleus, with their extensive innervation of limbic and higher brain regions and interactions with the endocrine system have important modulatory or regulatory effects on many cognitive, emotional and physiological processes. They have been strongly implicated in responses to stress and in the occurrence of major depressive disorder and other psychiatric disorders. In order to quantify some of these effects, detailed mathematical models of the activity of such cells are required which describe their complex neurochemistry and neurophysiology. We consider here a single-compartment model of these neurons which is capable of describing many of the known features of spike generation, particularly the slow rhythmic pacemaking activity often observed in these cells in a variety of species. Included in the model are 11 kinds of ion channels: a fast sodium current INa, a delayed rectifier potassium current IKDR, a transient potassium current IA, a slow non-inactivating potassium current IM, a low-threshold calcium current IT, two high threshold calcium currents IL and IN, small and large conductance potassium currents ISK and IBK, a hyperpolarization-activated cation current IH and a leak current ILeak. In Sections 3-8, each current type is considered in detail and parameters estimated from voltage clamp data where possible. Three kinds of model are considered for the BK current and two for the leak current. Intracellular calcium ion concentration Cai is an additional component and calcium dynamics along with buffering and pumping is discussed in Section 9. The remainder of the article contains descriptions of computed solutions which reveal both spontaneous and driven spiking with several parameter sets. Attention is focused on the properties usually associated with these neurons, particularly long duration of action potential, steep upslope on the leading edge of spikes, pacemaker-like spiking, long-lasting afterhyperpolarization

  1. 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 (United States)

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


    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

  2. The N Terminus of FliM Is Essential To Promote Flagellar Rotation in Rhodobacter sphaeroides


    Poggio, Sebastian; Osorio, Aurora; Corkidi, Gabriel; Dreyfus, Georges; Camarena, Laura


    FliM is part of the flagellar switch complex. Interaction of this protein with phospho-CheY (CheY-P) through its N terminus constitutes the main information relay point between the chemotactic system and the flagellum. In this work, we evaluated the role of the N terminus of FliM in the swimming behavior of Rhodobacter sphaeroides. Strains expressing the FliM protein with substitutions in residues previously reported in Escherichia coli as being important for interaction with CheY showed an i...

  3. 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.

  4. TrkB Signaling in Dorsal Raphe Nucleus is Essential for Antidepressant Efficacy and Normal Aggression Behavior. (United States)

    Adachi, Megumi; Autry, Anita E; Mahgoub, Melissa; Suzuki, Kanzo; Monteggia, Lisa M


    Brain-derived neurotrophic factor (BDNF) and its high affinity receptor, tropomyosin receptor kinase B (TrkB), have important roles in neural plasticity and are required for antidepressant efficacy. Studies examining the role of BDNF-TrkB signaling in depression and antidepressant efficacy have largely focused on the limbic system, leaving it unclear whether this signaling is important in other brain regions. BDNF and TrkB are both highly expressed in the dorsal raphe nucleus (DRN), a brain region that has been suggested to have a role in depression and antidepressant action, although it is unknown whether BDNF and TrkB in the dorsal raphe nucleus are involved in these processes. We combined the adeno-associated virus (AAV) with the Cre-loxP site-specific recombination system to selectively knock down either Bdnf or TrkB in the DRN. These mice were then characterized in several behavioral paradigms including measures of depression-related behavior and antidepressant efficacy. We show that knockdown of TrkB, but not Bdnf, in the DRN results in loss of antidepressant efficacy and increased aggression-related behavior. We also show that knockdown of TrkB or Bdnf in this brain region does not have an impact on weight, activity levels, anxiety, or depression-related behaviors. These data reveal a critical role for TrkB signaling in the DRN in mediating antidepressant responses and normal aggression behavior. The results also suggest a non-cell autonomous role for BDNF in the DRN in mediating antidepressant efficacy.

  5. 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: [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: [Institute of Chemistry, Academia Sinica, Taipei 115, Taiwan (China)


    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.

  6. Subaqueous terminus evolution at Tasman Glacier, New Zealand, as determined by remote-controlled survey (United States)

    Purdie, Heather; Bealing, Paul; Tidey, Emily; Harrison, Justin


    The presence of subaqueous ice ramps at the terminus of calving glaciers result from a combination of subaerial and subaqueous processes. These ice ramps eventually buoyantly calve, an event that can be hazardous to companies operating boat tours on proglacial lakes. However our knowledge of ice ramp forming processes, and feedbacks associated with their evolution, is sparse. We are using a remote controlled jet boat to survey bathymetry at an active calving margin. This vessel, mounted with both depth and side-scan sonar, can map subaqueous portions of the terminus right up to the active calving face at no risk to the operators. Surveys at the Tasman Glacier terminus over three consecutive years have revealed that subaqueous ice ramps are ephemeral features. In 2015 multiple ice ramps extended out into the lake from the terminus by 100-200 m, with the ramp surface being as much as 60 m below the water line at its outer perimeter. The maximum depth of the Tasman Lake at this time was 240 m. Within one month of the survey taking place, the largest of these ice ramps had calved and disintegrated. The consistent location of ice ramps between surveys indicates that other factors, like subglacial hydrology, may influence ice ramp evolution.

  7. Total Synthesis of Septocylindrin B and C-Terminus Modified Analogues (United States)

    Nelissen, Jo; Nuyts, Koen; De Zotti, Marta; Lavigne, Rob; Lamberigts, Chris; De Borggraeve, Wim M.


    The total synthesis is reported of the peptaibol Septocylindrin B which is related to the well documented channel forming peptaibol antibiotic Alamethicin. Several analogues were synthesized with a modified C-terminus, to investigate the SAR of the terminal residue Phaol. All these peptides were tested for their membrane perturbation properties by fluorescent dye leakage assay and for their antibacterial activity. PMID:23284749

  8. Total synthesis of Septocylindrin B and C-terminus modified analogues.

    Directory of Open Access Journals (Sweden)

    Jo Nelissen

    Full Text Available The total synthesis is reported of the peptaibol Septocylindrin B which is related to the well documented channel forming peptaibol antibiotic Alamethicin. Several analogues were synthesized with a modified C-terminus, to investigate the SAR of the terminal residue Phaol. All these peptides were tested for their membrane perturbation properties by fluorescent dye leakage assay and for their antibacterial activity.

  9. Analysis of the terminus region of the Caulobacter crescentus chromosome and identification of the dif site

    DEFF Research Database (Denmark)

    Jensen, Rasmus Bugge


    The terminus region of the Caulobacter crescentus chromosome and the dif chromosome dimer resolution site were characterized. The Caulobacter genome contains skewed sequences that abruptly switch strands at dif and may have roles in chromosome maintenance and segregation. Absence of dif or the Xer...

  10. 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.


    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...... of neurons in the midbrain raphe complex that projects to forebrain circuits regulating anxiety states, we used cholera toxin B subunit (CTb) as a retrograde tracer to identify neurons projecting to the basolateral amygdaloid complex (BL) in combination with c-Fos immunostaining to identify cells...... that activated neurons were serotonergic, non-serotonergic, or both. These data are consistent with the hypothesis that exposure to anxiogenic stimuli activates a subset of neurons in the midbrain raphe complex projecting to amygdala anxiety circuits Udgivelsesdato: 2008/12/10...

  11. Role of corticotropin-releasing factor in the median raphe nucleus in yohimbine-induced reinstatement of alcohol seeking in rats


    Lê, A.D.; Funk, Douglas; Coen, Kathleen; Li, Zhaoxia; Shaham, Yavin


    The pharmacological stressor yohimbine increases ongoing alcohol self-administration and reinstates alcohol seeking in rats. This effect is attenuated by systemic injections of a corticotropin-releasing factor (CRF) antagonist. The brain sites involved in CRF's role in yohimbine-induced alcohol taking and seeking are unknown. We report that injections of the CRF receptor antagonist d-Phe CRF into the median raphe nucleus (MRN) attenuated yohimbine-induced reinstatement of alcohol seeking but ...

  12. Differential role of serotonin projections from the dorsal and median raphe nuclei in phencyclidine-induced hyperlocomotion and fos-like immunoreactivity in rats. (United States)

    Kusljic, Snezana; Van Den Buuse, Maarten


    Altered brain serotonin activity is implicated in schizophrenia. We have previously shown differential involvement of serotonergic projections from the dorsal or median raphe nucleus in phencyclidine-induced hyperlocomotion in rats, a behavioral model of aspects of schizophrenia. Here we further investigated the effects of serotonergic lesions of the raphe nuclei on phencyclidine-induced hyperlocomotion by parallel assessment of Fos-like immunoreactivity (FLI), a marker of neuronal activation in the brain. Male Sprague-Dawley rats were anesthetized with pentobarbitone and stereotaxically microinjected with 5 μg of the serotonergic neurotoxin, 5,7-dihydroxytryptamine (5,7-DHT), into either the dorsal raphe (DRN) or median raphe nucleus (MRN). Two weeks after the surgery, rats with lesions of the MRN, but not those with lesions of the DRN, showed significant enhancement of the hyperlocomotion induced by injection of 2.5 mg/kg of phencyclidine. Rats with MRN lesions also showed significantly higher levels of FLI in the polymorphic layer of the dentate gyrus in the dorsal hippocampus (PoDG) when compared with sham-operated controls. Rats with lesions of the DRN showed significantly higher levels of FLI in the nucleus accumbens (NAcc). These results indicate that FLI in the PoDG, but not the NAcc, correlates with enhanced phencyclidine-induced locomotor hyperactivity in MRN-lesioned rats. These results support our previous studies suggesting a role of serotonergic projections from the MRN to the dorsal hippocampus in some of the symptoms of schizophrenia. Copyright © 2012 Wiley Periodicals, Inc.

  13. 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


    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

  14. Estradiol Valerate and Remifemin ameliorate ovariectomy-induced decrease in a serotonin dorsal raphe-preoptic hypothalamus pathway in rats. (United States)

    Wang, Wenjuan; Cui, Guangxia; Jin, Biao; Wang, Ke; Chen, Xing; Sun, Yu; Qin, Lihua; Bai, Wenpei


    Perimenopausal syndromes begin as ovarian function ceases and the most common symptoms are hot flushes. Data indicate that the projections of serotonin to hypothalamus may be involved in the mechanism of hot flushes. Therefore, the aim of this study is to investigate the potential role of the serotonin dorsal raphe-preoptic hypothalamus pathway for hot flushes in an animal model of menopause. We determined the changes in serotonin expression in the dorsal raphe (DR) and preoptic anterior hypothalamus (POAH) in ovariectomized rats. We also explored the therapeutical effects of estradiol valerate and Remifemin in this model. Eighty female Sprague-Dawley rats were randomly assigned to sham-operated (SHAM) group, ovariectomy (OVX) group with vehicle, ovariectomy with estradiol valerate treatment (OVX+E) group and ovariectomy with Remifemin (OVX+ICR) group. Serotonin expression was evaluated in the DR and POAH using immunofluorescence and quantified in the DR using an enzyme-linked immunosorbent assay (ELISA). Apoptosis was analyzed in the DR by TUNEL assay. The number of serotonin immunoreactive neurons and the level of serotonin expression in the DR decreased significantly following OVX compared to the SHAM group. No TUNEL-positive cells were detected in the DR in any group. In addition, following OVX, the number of serotonin-positive fibers decreased significantly in the ventromedial preoptic nucleus (VMPO), especially in the ventrolateral preoptic nucleus (VLPO). Treatment with either estradiol or Remifemin for 4 weeks countered the OVX-induced decreases in serotonin levels in both the DR and the hypothalamus, with levels in the treated rats similar to those in the SHAM group. A fluorescently labeled retrograde tracer was injected into the VLPO at the 4-week time point. A significantly lower percentage of serotonin with CTB double-labeled neurons in CTB-labeled neurons was demonstrated after ovariectomy, and both estradiol and Remifemin countered this OVX

  15. 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: [Myriad Genetic Laboratories Inc., 320 Wakara Way, Salt Lake City, UT 84108 (United States); Min, Jinrong, E-mail: [University of Toronto, 101 College Street, Toronto, ON M5G 1L7 (Canada); University of Toronto, Toronto, ON M5G 1L7 (Canada)


    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.

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    Medline Plus

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    Medline Plus

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  18. The Structure and Orientation of the C-Terminus of LRAP


    Shaw, Wendy J.; Ferris, Kim; Tarasevich, Barbara; Larson, Jenna L.


    Amelogenin is the predominant protein found during enamel development and is thought to be the biomineralization protein controlling the unique elongated hydroxyapatite crystals that constitute enamel. The secondary structure of biomineralization proteins is thought to be important in the interaction with hydroxyapatite. Unfortunately, very little data are available on the structure or the orientation of amelogenin, either in solution or bound to hydroxyapatite. The C-terminus contains the ma...

  19. The Cannabinoid Type-1 Receptor Carboxyl-Terminus, More Than Just a Tail (United States)

    Stadel, Rebecca; Ahn, Kwang H.; Kendall, Debra A.


    The cannabinoid type-1 (CB1) receptor is a G protein-coupled receptor (GPCR) that binds the main active ingredient of marijuana, Δ9-tetrahydrocannabinol, and has been implicated in several disease states, including drug addiction, anxiety, depression, obesity, and chronic pain. In the two decades since the discovery of CB1, studies at the molecular level have centered on the transmembrane core. This interest has now expanded as we discover that other regions of CB1, including the CB1 carboxyl-terminus, have critical structures that are important for CB1 activity and regulation. Following the recent description of the three dimensional structure of the full-length CB1 carboxyl-terminal tail (Ahn et al., Biopolymers (2009) 91: 565–573), several residues and structural motifs including two α-helices (termed H8 and H9) have been postulated to interact with common GPCR accessory proteins, such as G-proteins and β-arrestins. This discourse will focus on the CB1 carboxyl-terminus; our current understanding of the structural features of this region, evidence for its interaction with proteins, and the impact of structure on the binding and regulatory function of CB1 accessory proteins. The involvement of the carboxyl-terminus in the receptor life cycle including activation, desensitization, and internalization will be highlighted. PMID:21244428

  20. The hippocampus and dorsal raphe nucleus are key brain areas associated with the antidepressant effects of lithium augmentation of desipramine. (United States)

    Cussotto, Sofia; Cryan, John F; O'Leary, Olivia F


    Approximately 50% of depressed individuals fail to achieve remission with first-line antidepressant drugs and a third remain treatment-resistant. When first-line antidepressant treatment is unsuccessful, second-line strategies include dose optimisation, switching to another antidepressant, combination with another antidepressant, or augmentation with a non-antidepressant medication. Much of the evidence for the efficacy of augmentation strategies comes from studies using lithium to augment the effects of tricyclic antidepressants. The neural circuitry underlying the therapeutic effects of lithium augmentation is not yet fully understood. Recently, we reported that chronic treatment with a combination of lithium and the antidepressant desipramine, exerted antidepressant-like behavioural effects in a mouse strain (BALB/cOLaHsd) that did not exhibit an antidepressant-like behavioural response to either drug alone. In the present study, we used this model in combination with ΔFosB/FosB immunohistochemistry to identify brain regions chronically affected by lithium augmentation of desipramine when compared to either treatment alone. The data suggest that the dorsal raphe nucleus and the CA3 regions of the dorsal hippocampus are key nodes in the neural circuitry underlying antidepressant action of lithium augmentation of desipramine. These data give new insight into the neurobiology underlying the mechanism of lithium augmentation in the context of treatment-resistant depression. Copyright © 2017 Elsevier B.V. All rights reserved.

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


    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.

  2. 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


    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.

  3. Sleep deprivation reduces the citalopram-induced inhibition of serotoninergic neuronal firing in the nucleus raphe dorsalis of the rat. (United States)

    Prévot, E; Maudhuit, C; Le Poul, E; Hamon, M; Adrien, J


    Sleep deprivation (SD) for one night induces mood improvement in depressed patients. However, relapse often occurs on the day after deprivation subsequently to a sleep episode. In light of the possible involvement of central serotonin (5-hydroxytryptamine, 5-HT) neurotransmission in both depression and sleep mechanisms, we presently investigated, in the rat, the effects of SD and recovery sleep on the electrophysiological response of 5-HT neurons in the nucleus raphe dorsalis (NRD) to an acute challenge with the 5-HT reuptake blocker citalopram. In all rats, citalopram induced a dose-dependent inhibition of the firing of NRD neurons recorded under chloral hydrate anaesthesia. After SD, achieved by placing rats in a slowly rotating cylinder for 24 h, the inhibitory action of citalopram was significantly reduced (with a concomitant 53% increase in its ED50 value). After a recovery period of 4 h, a normal susceptibility of the firing to citalopram was restored. The decreased sensitivity of 5-HT neuronal firing to the inhibitory effect of citalopram after SD probably results in an enhancement of 5-HT neurotransmission. Such an adaptive phenomenon (similar to that reported after chronic antidepressant treatment), and its normalization after recovery sleep, parallel the mood improvement effect of SD and the subsequent relapse observed in depressed patients. These data suggest that the associated changes in 5-HT autocontrol of the firing of NRD serotoninergic neurons are relevant to the antidepressant action of SD.

  4. Monorail/Foxa2 regulates floorplate differentiation and specification of oligodendrocytes, serotonergic raphé neurones and cranial motoneurones (United States)

    Norton, Will H.; Mangoli, Maryam; Lele, Zsolt; Pogoda, Hans-Martin; Diamond, Brianne; Mercurio, Sara; Russell, Claire; Teraoka, Hiroki; Stickney, Heather L.; Rauch, Gerd-Jörg; Heisenberg, Carl-Philipp; Houart, Corinne; Schilling, Thomas F.; Frohnhoefer, Hans-Georg; Rastegar, Sepand; Neumann, Carl J.; Gardiner, R. Mark; Strähle, Uwe; Geisler, Robert; Rees, Michelle; Talbot, William S.; Wilson, Stephen W.


    Summary In this study, we elucidate the roles of the winged-helix transcription factor Foxa2 in ventral CNS development in zebrafish. Through cloning of monorail (mol), which we find encodes the transcription factor Foxa2, and phenotypic analysis of mol-/- embryos, we show that floorplate is induced in the absence of Foxa2 function but fails to further differentiate. In mol-/- mutants, expression of Foxa and Hh family genes is not maintained in floorplate cells and lateral expansion of the floorplate fails to occur. Our results suggest that this is due to defects both in the regulation of Hh activity in medial floorplate cells as well as cell-autonomous requirements for Foxa2 in the prospective laterally positioned floorplate cells themselves. Foxa2 is also required for induction and/or patterning of several distinct cell types in the ventral CNS. Serotonergic neurones of the raphé nucleus and the trochlear motor nucleus are absent in mol-/- embryos, and oculomotor and facial motoneurones ectopically occupy ventral CNS midline positions in the midbrain and hindbrain. There is also a severe reduction of prospective oligodendrocytes in the midbrain and hindbrain. Finally, in the absence of Foxa2, at least two likely Hh pathway target genes are ectopically expressed in more dorsal regions of the midbrain and hindbrain ventricular neuroepithelium, raising the possibility that Foxa2 activity may normally be required to limit the range of action of secreted Hh proteins. PMID:15677724

  5. The effect of continuous ELF-MFs on the level of 5-HIAA in the raphe nucleus of the rat. (United States)

    Shahbazi-Gahrouei, Daryoush; Shiri, Leila; Alaei, Hojjatollah; Naghdi, Naser


    The aim of this study was to investigate the effect of continuous extremely low frequency magnetic fields (ELF-MFs) with a frequency of 10 Hz and an intensity of 690-720 μT on the level of 5-hydroxyindolacetic acid (5-HIAA) in adult male Wistar rats. A total of 24 adult Wistar male rats were used, and after exposure with an ELF-MF for 15 successive days, all rats in each test were anesthetized with chloral hydrate. Then, they were placed in a stereotaxic frame for surgery and a microdialysis process. Dialysate samples were analyzed to measure the amount of 5-HIAA by high performance liquid chromatography (HPLC) using electrochemical detection. Results showed that ELF-MF exposure for 15 days, 1 h daily, was not effective in altering the level of 5-HIAA. However, ELF-MF exposure for 15 days, 3 h daily, decreased the level of the 5-HIAA in the raphe nucleus. It can be concluded that ELF-MFs affect the serotonergic system and may be used to treat nervous system diseases. This study is an initial step towards helping cure depression using ELF-MFs. © The Author 2016. Published by Oxford University Press on behalf of The Japan Radiation Research Society and Japanese Society for Radiation Oncology.

  6. The habenulo-raphe serotonergic circuit encodes an aversive expectation value essential for adaptive active avoidance of danger. (United States)

    Amo, Ryunosuke; Fredes, Felipe; Kinoshita, Masae; Aoki, Ryo; Aizawa, Hidenori; Agetsuma, Masakazu; Aoki, Tazu; Shiraki, Toshiyuki; Kakinuma, Hisaya; Matsuda, Masaru; Yamazaki, Masako; Takahoko, Mikako; Tsuboi, Takashi; Higashijima, Shin-ichi; Miyasaka, Nobuhiko; Koide, Tetsuya; Yabuki, Yoichi; Yoshihara, Yoshihiro; Fukai, Tomoki; Okamoto, Hitoshi


    Anticipation of danger at first elicits panic in animals, but later it helps them to avoid the real threat adaptively. In zebrafish, as fish experience more and more danger, neurons in the ventral habenula (vHb) showed tonic increase in the activity to the presented cue and activated serotonergic neurons in the median raphe (MR). This neuronal activity could represent the expectation of a dangerous outcome and be used for comparison with a real outcome when the fish is learning how to escape from a dangerous to a safer environment. Indeed, inhibiting synaptic transmission from vHb to MR impaired adaptive avoidance learning, while panic behavior induced by classical fear conditioning remained intact. Furthermore, artificially triggering this negative outcome expectation signal by optogenetic stimulation of vHb neurons evoked place avoidance behavior. Thus, vHb-MR circuit is essential for representing the level of expected danger and behavioral programming to adaptively avoid potential hazard. Copyright © 2014 Elsevier Inc. All rights reserved.

  7. The role of lateral habenula-dorsal raphe nucleus circuits in higher brain functions and psychiatric illness. (United States)

    Zhao, Hua; Zhang, Bei-Lin; Yang, Shao-Jun; Rusak, Benjamin


    Serotonergic neurons in the dorsal raphe nucleus (DRN) play an important role in regulation of many physiological functions. The lateral nucleus of the habenular complex (LHb) is closely connected to the DRN both morphologically and functionally. The LHb is a key regulator of the activity of DRN serotonergic neurons, and it also receives reciprocal input from the DRN. The LHb is also a major way-station that receives limbic system input via the stria medullaris and provides output to the DRN and thereby indirectly connects a number of other brain regions to the DRN. The complex interactions of the LHb and DRN contribute to the regulation of numerous important behavioral and physiological mechanisms, including those regulating cognition, reward, pain sensitivity and patterns of sleep and waking. Disruption of these functions is characteristic of major psychiatric illnesses, so there has been a great deal of interest in how disturbed LHb-DRN interactions may contribute to the symptoms of these illnesses. This review summarizes recent research related to the roles of the LHb-DRN system in regulation of higher brain functions and the possible role of disturbed LHb-DRN function in the pathogenesis of psychiatric disorders, especially depression. Copyright © 2014 Elsevier B.V. All rights reserved.

  8. Viral vector mediated expression of mutant huntingtin in the dorsal raphe produces disease-related neuropathology but not depressive-like behaviors in wildtype mice. (United States)

    Pitzer, Mark; Lueras, Jordan; Warden, Anna; Weber, Sydney; McBride, Jodi


    Huntington׳s disease (HD) is a neurodegenerative disorder caused by a mutation in the HTT gene (mHTT) encoding the protein huntingtin. An expansion in the gene׳s CAG repeat length renders a misfolded, dysfunctional protein with an abnormally long glutamine (Q) stretch at the N terminus that often incorporates into inclusion bodies and leads to neurodegeneration in many regions of the brain. HD is characterized by motor and cognitive decline as well as mood disorders, with depression being particularly common. Approximately 40% of the HD population suffers from depressive symptoms. Because these symptoms often manifest a decade or more prior to the knowledge that the person is at risk for the disease, a portion of the early depression in HD appears to be a consequence of the pathology arising from expression of the mutant gene. While the depression in HD patients is often treated with serotonin agonists, there is scant experimental evidence that the depression in HD responds well to these serotonin treatments or in a similar manner to how non-HD depression tends to respond. Additionally, at very early sub-threshold depression levels, abnormal changes in several neuronal populations are already detectable in HD patients, suggesting that a variety of brain structures may be involved. Taken together, the serotonin system is a viable candidate. However, at present there is limited evidence of the precise nuclei or circuits that play a role in HD depression. With this in mind, the current study was designed to control for the widespread brain neuropathology that occurs in HD and in transgenic mouse models of HD and focuses specifically on the influence of the midbrain dorsal raphe nucleus (DRN). The DRN provides the majority of the serotonin to the forebrain and exhibits cell loss in non-HD depression. Therefore, we employed a viral vector delivery system to investigate whether the over-expression of mHTT in the DRN׳s ventral sub-nuclei alone is sufficient to produce

  9. 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.

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  12. Computational Analysis of the CB1 Carboxyl-terminus in the Receptor-G Protein Complex


    Shim, Joong-Youn; Khurana, Leepakshi; Kendall, Debra A.


    Despite the important role of the carboxyl-terminus (Ct) of the activated brain cannabinoid receptor one (CB1) in the regulation of G protein signaling, a structural understanding of interactions with G proteins is lacking. This is largely due to the highly flexible nature of the CB1 Ct that dynamically adapts its conformation to the presence of G proteins. In the present study, we explored how the CB1 Ct can interact with the G protein by building on our prior modeling of the CB1-Gi complex ...

  13. Dorsal-to-Ventral Shift in Midbrain Dopaminergic Projections and Increased Thalamic/Raphe Serotonergic Function in Early Parkinson Disease. (United States)

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


    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.

  14. Presynaptic α4β2 nicotinic acetylcholine receptors increase glutamate release and serotonin neuron excitability in the dorsal raphe nucleus. (United States)

    Garduño, Julieta; Galindo-Charles, Luis; Jiménez-Rodríguez, Javier; Galarraga, Elvira; Tapia, Dagoberto; Mihailescu, Stefan; Hernandez-Lopez, Salvador


    Several behavioral effects of nicotine are mediated by changes in serotonin (5-HT) release in brain areas that receive serotonergic afferents from the dorsal raphe nucleus (DRN). In vitro experiments have demonstrated that nicotine increases the firing activity in the majority of DRN 5-HT neurons and that DRN contains nicotinic acetylcholine receptors (nAChRs) located at both somata and presynaptic elements. One of the most common presynaptic effects of nicotine is to increase glutamate release. Although DRN receives profuse glutamatergic afferents, the effect of nicotine on glutamate release in the DRN has not been studied in detail. Using whole-cell recording techniques, we investigated the effects of nicotine on the glutamatergic input to 5-HT DRN neurons in rat midbrain slices. Low nicotine concentrations, in the presence of bicuculline and tetrodotoxin (TTX), increased the frequency but did not change the amplitude of glutamate-induced EPSCs, recorded from identified 5-HT neurons. Nicotine-induced increase of glutamatergic EPSC frequency persisted 10-20 min after drug withdrawal. This nicotinic effect was mimicked by exogenous administration of acetylcholine (ACh) or inhibition of ACh metabolism. In addition, the nicotine-induced increase in EPSC frequency was abolished by blockade of α4β2 nAChRs, voltage-gated calcium channels, or intracellular calcium signaling but not by α7 nAChR antagonists. These data suggest that both nicotine and endogenous ACh can increase glutamate release through activation of presynaptic α4β2 but not α7 nAChRs in the DRN. The effect involves long-term changes in synaptic function, and it is dependent on voltage-gated calcium channels and presynaptic calcium stores.

  15. GABAA receptors in the dorsal raphé nucleus of mice: escalation of aggression after alcohol consumption (United States)

    Takahashi, Aki; Kwa, Carolyn; DeBold, Joseph F.


    Rationale The dorsal raphé nucleus (DRN), the origin for serotonin (5-HT) in forebrain areas, has been implicated in the neural control of escalated aggression. Gamma aminobutyric acid type-A (GABAA) and type-B (GABAB) receptors are expressed in the DRN and modulate 5-HT neuronal activity, and both play a role in the behavioral effect of alcohol. Objective The purpose of this study is to examine the interaction between drugs acting on GABA receptors in the DRN and alcohol in their effects on aggressive behaviors. Method Male CFW mice, housed with a female, were trained to self-administer ethanol (1.0 g/kg) or water via an operant conditioning panel in their home cage. Immediately after they drank either ethanol or water, the animals were microinfused with a GABAergic drug into the DRN, and their aggressive behaviors were assessed 10 min later. Muscimol (0.006 nmol), a GABAA receptor agonist, escalated alcohol-heightened aggression but had no effect in the absence of ethanol. This effect of muscimol was prominent in the animals that showed alcohol-heightened aggression, but not the animals that reduced or did not change aggressive behavior after ethanol infusion compared to water. On the other hand, the GABAB agonist baclofen (0.06 nmol) increased aggressive behavior similarly in both water and ethanol conditions. Antagonists of the GABAA and GABAB receptors, bicuculline (0.006 nmol) and phaclofen (0.3 nmol) respectively, did not suppress heightened-aggressive behavior induced by ethanol self-administration. Conclusion GABAA receptors in the DRN are one of the neurobiological targets of alcohol-heightened aggression. Activation of the GABAB receptors in the DRN also produced escalated aggression, but that is independent of the effect of alcohol. PMID:20589493

  16. GABA(A) receptors in the dorsal raphé nucleus of mice: escalation of aggression after alcohol consumption. (United States)

    Takahashi, Aki; Kwa, Carolyn; Debold, Joseph F; Miczek, Klaus A


    The dorsal raphé nucleus (DRN), the origin for serotonin (5-HT) in forebrain areas, has been implicated in the neural control of escalated aggression. Gamma aminobutyric acid type-A (GABA(A)) and type-B (GABA(B)) receptors are expressed in the DRN and modulate 5-HT neuronal activity, and both play a role in the behavioral effect of alcohol. The purpose of this study is to examine the interaction between drugs acting on GABA receptors in the DRN and alcohol in their effects on aggressive behaviors. Male CFW mice, housed with a female, were trained to self-administer ethanol (1.0 g/kg) or water via an operant conditioning panel in their home cage. Immediately after they drank either ethanol or water, the animals were microinfused with a GABAergic drug into the DRN, and their aggressive behaviors were assessed 10 min later. Muscimol (0.006 nmol), a GABA(A) receptor agonist, escalated alcohol-heightened aggression but had no effect in the absence of ethanol. This effect of muscimol was prominent in the animals that showed alcohol-heightened aggression, but not the animals that reduced or did not change aggressive behavior after ethanol infusion compared to water. On the other hand, the GABA(B) agonist baclofen (0.06 nmol) increased aggressive behavior similarly in both water and ethanol conditions. Antagonists of the GABA(A) and GABA(B) receptors, bicuculline (0.006 nmol) and phaclofen (0.3 nmol) respectively, did not suppress heightened-aggressive behavior induced by ethanol self-administration. GABA(A) receptors in the DRN are one of the neurobiological targets of alcohol-heightened aggression. Activation of the GABA(B) receptors in the DRN also produced escalated aggression, but that is independent of the effect of alcohol.

  17. Effects of sleep deprivation on serotonergic neuronal activity in the dorsal raphe nucleus of the freely moving cat. (United States)

    Gardner, J P; Fornal, C A; Jacobs, B L


    Total sleep deprivation (TSD) for one or more nights produces a rapid antidepressant response in humans. Since most pharmacological treatments for depression increase brain serotonin neurotransmission, the purpose of the present study was to determine whether TSD increases the activity of serotonergic neurons in the dorsal raphe nucleus (DRN) in cats. Cats were prevented from sleeping by the experimenter, who monitored the behavioral state of each animal on a polygraph. Firing rates during quiet waking (QW) and active waking (AW) were obtained throughout a 24-h sleep deprivation period and subsequent 6-h recovery period. During the experiments, unit activity was also recorded during exposure to loud white noise, which elicited strong behavioral arousal. The inhibitory response of serotonergic DRN neurons to systemic administration of the selective 5-HT1A agonist 8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT) was determined before and after TSD to assess possible changes in 5-HT1A autoreceptor sensitivity. TSD increased mean firing rates by as much as 18% during both AW and white noise exposure. Maximal effects were observed after 15 h of TSD for AW, and after 18 h for white noise. QW firing rates also tended to be elevated throughout TSD. Firing rates for all conditions during the recovery period were not significantly different from baseline. The neuronal inhibition produced by 8-OH-DPAT was significantly diminished after TSD. Overall, these results indicate that TSD increases the firing rate of serotonergic DRN neurons during AW and arousal. This effect may be attributable to a decrease in the sensitivity of 5-HT1A autoreceptors. These findings are consistent with the hypothesis that TSD exerts its antidepressant action, at least in part, through an activation of brain serotonergic neurons.

  18. Cellular adaptations of dorsal raphe serotonin neurons associated with the development of active coping in response to social stress. (United States)

    Wood, Susan K; Zhang, Xiao-Yan; Reyes, Beverly A S; Lee, Catherine S; Van Bockstaele, Elisabeth J; Valentino, Rita J


    Social stress is a risk factor for affective disorders for certain vulnerable individuals. Stress and depression are linked in part through regulation of the dorsal raphe (DR)-serotonin (5-HT) system by the stress-related neuropeptide, corticotropin-releasing factor (CRF). We used a rat social stress model that shows individual differences in coping strategies to determine whether differences in CRF-5-HT interactions underlie individual differences in the vulnerability to social stress. Rats were exposed to the resident-intruder model of social stress for 5 days. In vivo single-unit recordings assessed DR-5-HT neuronal responses to CRF and immunoelectron microscopy assessed CRF1 and CRF2 cellular localization 24 hours after the last stress. Rats responded to social stress passively, assuming defeat with short latencies (48%), or actively, with proactive behaviors and longer defeat latencies (LL, 52%). Whereas CRF (30 ng, intra-DR) inhibited 5-HT neuronal activity of control and SL rats, it activated 5-HT neurons of LL rats, an effect that was CRF2-mediated. Consistent with this, social stress promoted CRF1 internalization together with CRF2 recruitment to the plasma membrane of DR neurons selectively in LL rats. These data suggest that a proactive coping strategy toward social stress is associated with a redistribution of CRF1 and CRF2 in DR-5-HT neurons that primes the system to be activated by subsequent stress. The lack of this adaptation in passive coping rats may contribute to their depressive-like phenotype. These studies provide a cellular mechanism for individual differences in stress responses and consequences. Copyright © 2013 Society of Biological Psychiatry. Published by Elsevier Inc. All rights reserved.

  19. Median raphe stimulation-induced motor inhibition concurrent with suppression of type 1 and type 2 hippocampal theta. (United States)

    Bland, Brian H; Bland, Cheryl E; MacIver, M Bruce


    This study investigated behavioral, anatomical and electrophysiological effects produced by electrical stimulation of posterior hypothalamic (PH) or median raphe (MR) nuclei, independently and during combined stimulation of both PH and MR. These three stimulation conditions were applied during spontaneous behavior in an open field and during PH stimulation-induced wheel running, while simultaneously recording hippocampal (HPC) field activity. An additional objective was to determine the effects of MR stimulation on Type 1 movement related theta and Type 2 sensory processing related theta. To achieve the latter, when behavioral studies were completed we studied the same rats under urethane anesthesia and then during urethane anesthesia with the addition of atropine sulfate (ATSO4). Here we demonstrated that electrical stimulation of a localized region of the MR nucleus resulted in a profound inhibition of both spontaneously occurring theta related motor behaviors and the theta related motor behaviors induced by electrical stimulation of the PH nucleus. Furthermore, this motor inhibition occurred concurrently with strong suppression of hippocampal theta field oscillations in the freely moving rat, a condition where the theta recorded is Type 2 sensory processing theta occurring coincidently with Type 1 movement related theta (Bland, 1986). Our results indicate that motor inhibition resulted from stimulation of neurons located in the mid central region of the MR, while stimulation in adjacent regions produced variable responses, including movements and theta activity. The present study provided evidence that the pharmacological basis of the suppression of Type 2 sensory processing HPC theta was cholinergic. However, MR inhibition of PH-induced wheel running was not affected by cholinergic blockade, which blocks Type 2 theta, indicating that MR stimulation-induced motor inhibition also requires the suppression of Type 1 theta. © 2015 Wiley Periodicals, Inc.

  20. Cytomorphometric changes in the dorsal raphe neurons after rapid eye movement sleep deprivation are mediated by noradrenalin in rats

    Directory of Open Access Journals (Sweden)

    Biswas Sudipta


    Full Text Available Abstract Objectives This study was carried out to investigate the effect of rapid eye movement sleep (REMS deprivation (REMSD on the cytomorphology of the dorsal raphe (DR neurons and to evaluate the possible role of REMSD-induced increased noradrenalin (NA in mediating such effects. Methods Rats were REMS deprived by the flowerpot method; free moving normal home cage rats, large platform and post REMS-deprived recovered rats were used as controls. Further, to evaluate if the effects were induced by NA, separate sets of experimental rats were treated (i.p. with α1-adrenoceptor antagonist, prazosin (PRZ. Histomorphometric analysis of DR neurons in stained brain sections were performed in experimental and control rats; neurons in inferior colliculus (IC served as anatomical control. Results The mean size of DR neurons was larger in REMSD group compared to controls, whereas, neurons in the recovered group of rats did not significantly differ than those in the control animals. Further, mean cell size in the post-REMSD PRZ-treated animals was comparable to those in the control groups. IC neurons were not affected by REMSD. Conclusions REMS loss has been reported to impair several physiological, behavioral and cellular processes. The mean size of the DR neurons was larger in the REMS deprived group of rats than those in the control groups; however, in the REMS deprived and prazosin treated rats the size was comparable to the normal rats. These results showed that REMSD induced increase in DR neuronal size was mediated by NA acting on α1-adrenoceptor. The findings suggest that the sizes of DR neurons are sensitive to REMSD, which if not compensated could lead to neurodegeneration and associated disorders including memory loss and Alzheimer's disease.

  1. Cannabidiol inhibits the reward-facilitating effect of morphine: involvement of 5-HT1A receptors in the dorsal raphe nucleus. (United States)

    Katsidoni, Vicky; Anagnostou, Ilektra; Panagis, George


    Cannabidiol is a non-psychotomimetic constituent of Cannabis sativa, which induces central effects in rodents. It has been shown that cannabidiol attenuates cue-induced reinstatement of heroin seeking. However, to the best of our knowledge, its effects on brain stimulation reward and the reward-facilitating effects of drugs of abuse have not yet been examined. Therefore, we investigated the effects of cannabidiol on brain reward function and on the reward-facilitating effect of morphine and cocaine using the intracranial self-stimulation (ICSS) paradigm. Rats were prepared with a stimulating electrode into the medial forebrain bundle (MFB), and a guide cannula into the dorsal raphe (microinjection experiments), and were trained to respond for electrical brain stimulation. A low dose of cannabidiol did not affect the reinforcing efficacy of brain stimulation, whereas higher doses significantly elevated the threshold frequency required for MFB ICSS. Both cocaine and morphine lowered ICSS thresholds. Cannabidiol inhibited the reward-facilitating effect of morphine, but not cocaine. This effect was reversed by pre-treatment with an intra-dorsal raphe injection of the selective 5-HT1A receptor antagonist WAY-100635. The present findings indicate that cannabidiol does not exhibit reinforcing properties in the ICSS paradigm at any of the doses tested, while it decreases the reward-facilitating effects of morphine. These effects were mediated by activation of 5-HT1A receptors in the dorsal raphe. Our results suggest that cannabidiol interferes with brain reward mechanisms responsible for the expression of the acute reinforcing properties of opioids, thus indicating that cannabidiol may be clinically useful in attenuating the rewarding effects of opioids. © 2012 The Authors, Addiction Biology © 2012 Society for the Study of Addiction.

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

    DEFF Research Database (Denmark)

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


    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...

  3. 5-HT(1A) receptor binding in the dorsal raphe nucleus is implicated in the anxiolytic-like effects of Cinnamomum cassia. (United States)

    Jung, Yang-Hee; Kwon, Seung-Hwan; Hong, Sa-Ik; Lee, Sung-Ok; Kim, Sun-Yeou; Lee, Seok-Yong; Jang, Choon-Gon


    Previously we reported that the 50% EtOH extract of Cinnamomum cassia (C. cassia) possesses anxiolytic-like activity in the mouse elevated plus maze (EPM) test. This activity was blocked by the 5-HT(1A) receptor antagonist, WAY 100635. Therefore, in order to investigate the effect of C. cassia on 5-HT(1A) receptor binding, quantitative autoradiography of 5-HT(1A) receptors was carried out in brains of mice treated acutely and repeatedly with C. cassia. Binding of [(3)H]8-OH-DPAT to the 5-HT(1A) receptor was investigated in the mouse brain. After a single treatment of C. cassia (750 mg/kg, p.o.), [(3)H]8-OH-DPAT binding showed a significant increase in the dorsal raphe nucleus (DRN). After repeated treatment with C. cassia (100mg/kg, once a day for 5 days, p.o.), [(3)H]8-OH-DPAT binding showed no significant change in any brain region. Taken together, the anxiolytic-like effect of the 50% EtOH extract of C. cassia might be mediated by region specific change of 5-HT(1A) receptors in the dorsal raphe nucleus. Copyright © 2012. Published by Elsevier Inc.

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

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


    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

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

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


    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. 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


    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...... on GPCR-to-effector specificity. Dually modified Galpha proteins were also superior in conferring high-affinity agonist sites onto a coexpressed GPCR in the absence, but not in the presence, of guanine nucleotides. Together, our data suggest that receptor-G protein coupling selectivity involves...

  7. Create Your Plate

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  8. 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 ...

  9. Create Your Plate

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    Full Text Available ... Share: Print Page Text Size: A A A Listen En Español Create Your Plate Create Your Plate is a simple and effective way to manage your blood glucose levels and lose weight. With this method, you fill your plate with more non-starchy veggies ...

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

    Directory of Open Access Journals (Sweden)

    Toratane Munegumi


    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.

  11. 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


    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...... terminus in mediating the action of amphetamines by focusing initially on the highly conserved threonine residue at position 81, a candidate site for phosphorylation by protein kinase C. Molecular dynamics simulations of the wild type SERT, compared with its mutations SERT(T81A) and SERT(T81D), suggested......, SERT(T81A) (and the homologous mutations in noradrenaline and dopamine) failed to support amphetamine-induced efflux, and this was not remedied by aspartate at this position. Amphetamine-induced currents through SERT(T81A) were comparable with those through the wild type transporter. Both abundant Na...

  12. Carboxyl terminus heterogeneity of type IV fimbrial subunit protein of Pasteurella multocida isolates. (United States)

    Shivachandra, Sathish Bhadravati; Kumar, Abhinendra; Yogisharadhya, Revanaiah; Ramakrishnan, M A; Viswas, K N


    Pasteurella multocida, a Gram-negative bacterial pathogen, known to affect a wide range of domestic as well as wild animal and avian species throughout the world by causing either systemic or localized infections termed as 'pasteurellosis'. P. multocida isolates are known to possess type IV fimbriae (pili) as one of the major virulence factors based on their role in adhesion to host surfaces and subsequent pathogenesis. In the present study, ptfA gene of Indian P. multocida isolates (n = 8) originated from different animal (buffalo, sheep, goat, pig) and avian host species (chicken, turkey, duck, quail) were amplified, cloned, sequenced and compared with available ptfA/fimbrial protein sequences in GenBank/publications (n = 22) to understand its variability with respect to geography/host/serogroup/disease specific patterns. Multiple sequence alignment revealed highly conserved N-terminus α-1 helix region and heterogeneous C-terminus (68-137 aa) comprised of β-strand regions (β1, β2, β3, β4) with conserved two pairs of cysteine residues. Interestingly, an existence of absolute homogeneity among the P. multocida isolates that caused haemorrhagic septicaemia in bovines and septicaemic pasteurellosis in sheep and goats was noticed. Pig isolates had 99.3% homogeneity. On contrary, more diversity (35.8%) was observed among isolates that caused fowl cholera in avians irrespective of identical capsular/somatic serogroup and similar host species. Phylogenetic analysis based on nucleotide sequences of ptfA gene revealed formation of mixed clusters with isolates representing different disease conditions as well as serogroups irrespective of country of origin which indicated the possible role of cross-species transmission among different animal/avian species. The study indicated highly conserved and host specific fimbriae among animal species than relatively divergent fimbriae among avian species.

  13. The N-terminus of GalE induces tmRNA activity in Escherichia coli. (United States)

    Ruhe, Zachary C; Hayes, Christopher S


    The tmRNA quality control system recognizes stalled translation complexes and facilitates ribosome recycling in a process termed 'ribosome rescue'. During ribosome rescue, nascent chains are tagged with the tmRNA-encoded SsrA peptide, which targets tagged proteins for degradation. In Escherichia coli, tmRNA rescues ribosomes arrested on truncated messages, as well as ribosomes that are paused during elongation and termination. Here, we describe a new translational pausing determinant that leads to SsrA peptide tagging of the E. coli GalE protein (UDP-galactose 4-epimerase). GalE chains are tagged at more than 150 sites, primarily within distinct clusters throughout the C-terminal domain. These tagging sites do not correspond to rare codon clusters and synonymous recoding of the galE gene had little effect on tagging. Moreover, tagging was largely unaffected by perturbations that either stabilize or destabilize the galE transcript. Examination of GalE-thioredoxin (TrxA) fusion proteins showed that the GalE C-terminal domain is no longer tagged when fused to an N-terminal TrxA domain. Conversely, the N-terminus of GalE induced tagging within the fused C-terminal TrxA domain. These findings suggest that translation of the GalE N-terminus induces subsequent tagging of the C-terminal domain. We propose that co-translational maturation of the GalE N-terminal domain influences ribosome pausing and subsequent tmRNA activity.

  14. Creating more effective graphs

    CERN Document Server

    Robbins, Naomi B


    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

  15. Create Your Plate

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    Full Text Available ... Health Care Professionals Law Enforcement Driver's License For Lawyers Food & Fitness Home Food MyFoodAdvisor Recipes Association Cookbook Recipes Planning Meals Diabetes Meal Plans Create Your Plate Gluten ...

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    Full Text Available ... Care Blood Glucose Testing Medication Doctors, Nurses & More Oral Health & Hygiene Women A1C Insulin Pregnancy 8 Tips for ... Diabetes Meal Plans Create Your Plate Gluten Free Diets Meal Planning for Vegetarian Diets Cook with Heart- ...

  17. Create Your Plate (United States)

    ... 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 ...

  18. 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 ...

  19. Create Your Plate

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    Full Text Available ... Pacific Islanders American Indian/Alaska Native Programs Older Adults Family Link Diabetes EXPO Upcoming Diabetes EXPOs EXPO ... Plate! Click on the plate sections below to add your food choices. Reset Plate Share Create Your ...

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    Full Text Available ... of Diabetes Research & Practice Home We Are Research Leaders World's Largest Diabetes Meeting Recent Advances Type 1 ... Your Plate It's simple and effective for both managing diabetes and losing weight. Creating your plate lets ...

  1. Create Your Plate

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    Full Text Available ... Now - match-donate-now.html Match – Donate Now Make your year-end donation today and ... Tour Registration Is Open It starts with you. Sign up to ride in Tour de Cure and create ...

  2. Create Your Plate

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    Full Text Available ... this interactive tool. The healthy meal combinations are endless! Ready to try it at home? Just follow ... non-starchy vegetables and that your options are endless. Create Your Plate! Click on the plate sections ...

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    Full Text Available ... and Pacific Islanders American Indian/Alaska Native Programs Older Adults Family Link Diabetes EXPO Upcoming Diabetes EXPOs ... Planning Meals Diabetes Meal Plans and a Healthy Diet Create Your Plate Meal Planning for Vegetarian Diets ...

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  9. Creating unreal audio


    Rudsengen, Mathias Flaten


    Creating unreal audio” refers to the act of designing a sound effect that is intended to sound like a completely fictional object. This thesis is a practical venture into digital audio design. During the process of creating a sound effect anchored in a specific thematic framework, I will describe my work process and the challenges and problems faced, showing my personal work process and how modern digital sound effect creation can be undertaken. To provide context, I will also describe and re...

  10. Detection of ALMB-toxin in the larval body of Myrmeleon bore by anti-N-terminus peptide antibodies. (United States)

    Yoshida, N; Sugama, H; Gotoh, S; Matsuda, K; Nishimura, K; Komai, K


    Antibodies were raised against a synthetic antigen carrying the N-terminus peptide of ALMB-toxin, which had been isolated from the antlion, Myrmeleon bore, that exhibited high specificity to the toxin. Analyses with the antibodies showed the toxin to be present mainly at the larval stage and localized in a region from the thorax to abdomen of the larval body.

  11. Harvey murine sarcoma virus p21 ras protein: biological and biochemical significance of the cysteine nearest the carboxy terminus

    DEFF Research Database (Denmark)

    Willumsen, B M; Norris, K; Papageorge, A G


    Previous studies of premature chain termination mutants and in frame deletion mutants of the p21 ras transforming protein encoded by the transforming gene of Harvey murine sarcoma virus (Ha-MuSV) have suggested that the C terminus is required for cellular transformation, lipid binding, and membrane...

  12. Structural and functional analysis of a bipolar replication terminus - Implications for the origin of polarity of fork arrest

    NARCIS (Netherlands)

    Mohanty, BK; Bussiere, DE; Sahoo, T; Pai, KS; Meijer, WJJ; Bron, S; Bastia, D


    We have delineated the amino acid to nucleotide contacts made by two interacting dimers of the replication terminator protein (RTP) of Bacillus subtilis with a novel naturally occurring bipolar replication terminus by converting RTP to a site-directed chemical nuclease and mapping its cleavage sites

  13. 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: [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)


    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

  14. Creating a learning culture. (United States)

    Mathewson, Karyn


    This column describes the efforts of an agency to build a learning culture as part of changing their approach to service delivery, when adopting a focus on psychiatric rehabilitation and recovery. This example of one organization's challenges and successes in workforce development provides an alternative approach to the common single-session staff training that typically fails to change practice. This description draws from published material on communities of practice, technical consultation, and agency experience. Training alone is not enough to create change. An organizational commitment to ongoing quality improvement, along with available and accessible technical assistance for staff, creates an environment where change is anticipated, managed, and celebrated.

  15. Creating Web Pages Simplified

    CERN Document Server

    Wooldridge, Mike


    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

  16. Disturbances in the FGFR1-5-HT1A Heteroreceptor Complexes in the Raphe-Hippocampal 5-HT System Develop in a Genetic Rat Model of Depression

    Directory of Open Access Journals (Sweden)

    Dasiel O. Borroto-Escuela


    Full Text Available The FGFR1-5-HT1A heteroreceptor complexes are involved in neuroplasticity in the rat hippocampus and in the mesencephalic raphe 5-HT nerve cells. There exists a 5-HT1A protomer enhancement of FGFR1 protomer signaling. Acute and 10 day treatment with intracerebroventricular (i.c.v. FGF-2 and the 5-HT1A agonist 8-OH-DPAT produced enhanced antidepressant effects in the forced swim test (FST. We studied in the current work the disturbances in the FGFR1-5-HT1A heterocomplexes in a genetic rat model of depression, the Flinders sensitive line (FSL rats of Sprague-Dawley (SD origin, by means of neurochemical, neurophysiological and behavioral techniques. In control SD rats, the FGFR1 agonist SUN11602 and FGF2 produced a significant reduction of G protein-coupled inwardly rectifying K+ channel (GIRK currents induced by 8-OH-DPAT in the CA1 area of the hippocampus. In FSL rats, only i.c.v. 8-OH-DPAT alone treatment produced a significant reduction in the immobility time. The combined i.c.v. treatment (FGF2 + 8-OH-DPAT in FSL rats did not cause a significant decrease in immobility time in the FST. However, in the SD rats this combined treatment produced a significant reduction. Furthermore, in the FSL rat a significant increase in the density of FGFR1-5-HT1A proximity ligation assay (PLA positive clusters was only found after i.c.v. 8-OH-DPAT treatment alone in the CA2 and CA3 areas. In the SD rat a significant increase in the density of specific PLA clusters was only observed in the CA2 area of the i.c.v. combined treatment (FGF2 + 8-OH-DPAT group. No treatment led to significant changes in the PLA clusters of the dorsal raphe in the FSL rat. However, significant changes in the density of specific PLA clusters were only found in the dorsal raphe of SD rats after combined treatment and treatment with 8-OH-DPAT alone. The results indicate that in FSL rats compared with SD rats alterations may develop in the ability of 8-OH-DPAT and combined FGFR1 and 5-HT1A

  17. 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.


    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.

  18. Disturbances in the FGFR1-5-HT1A Heteroreceptor Complexes in the Raphe-Hippocampal 5-HT System Develop in a Genetic Rat Model of Depression. (United States)

    Borroto-Escuela, Dasiel O; DuPont, Caitlin M; Li, Xiang; Savelli, David; Lattanzi, Davide; Srivastava, Ipsit; Narváez, Manuel; Di Palma, Michael; Barbieri, Elisa; Andrade-Talavera, Yuniesky; Cuppini, Riccardo; Odagaki, Yuji; Palkovits, Miklos; Ambrogini, Patrizia; Lindskog, Maria; Fuxe, Kjell


    The FGFR1-5-HT1A heteroreceptor complexes are involved in neuroplasticity in the rat hippocampus and in the mesencephalic raphe 5-HT nerve cells. There exists a 5-HT1A protomer enhancement of FGFR1 protomer signaling. Acute and 10 day treatment with intracerebroventricular (i.c.v.) FGF-2 and the 5-HT1A agonist 8-OH-DPAT produced enhanced antidepressant effects in the forced swim test (FST). We studied in the current work the disturbances in the FGFR1-5-HT1A heterocomplexes in a genetic rat model of depression, the Flinders sensitive line (FSL) rats of Sprague-Dawley (SD) origin, by means of neurochemical, neurophysiological and behavioral techniques. In control SD rats, the FGFR1 agonist SUN11602 and FGF2 produced a significant reduction of G protein-coupled inwardly rectifying K(+) channel (GIRK) currents induced by 8-OH-DPAT in the CA1 area of the hippocampus. In FSL rats, only i.c.v. 8-OH-DPAT alone treatment produced a significant reduction in the immobility time. The combined i.c.v. treatment (FGF2 + 8-OH-DPAT) in FSL rats did not cause a significant decrease in immobility time in the FST. However, in the SD rats this combined treatment produced a significant reduction. Furthermore, in the FSL rat a significant increase in the density of FGFR1-5-HT1A proximity ligation assay (PLA) positive clusters was only found after i.c.v. 8-OH-DPAT treatment alone in the CA2 and CA3 areas. In the SD rat a significant increase in the density of specific PLA clusters was only observed in the CA2 area of the i.c.v. combined treatment (FGF2 + 8-OH-DPAT) group. No treatment led to significant changes in the PLA clusters of the dorsal raphe in the FSL rat. However, significant changes in the density of specific PLA clusters were only found in the dorsal raphe of SD rats after combined treatment and treatment with 8-OH-DPAT alone. The results indicate that in FSL rats compared with SD rats alterations may develop in the ability of 8-OH-DPAT and combined FGFR1 and 5-HT1A agonist

  19. Role of corticotropin-releasing factor in the median raphe nucleus in yohimbine-induced reinstatement of alcohol seeking in rats. (United States)

    Lê, A D; Funk, Douglas; Coen, Kathleen; Li, Zhaoxia; Shaham, Yavin


    The pharmacological stressor yohimbine increases ongoing alcohol self-administration and reinstates alcohol seeking in rats. This effect is attenuated by systemic injections of a corticotropin-releasing factor (CRF) antagonist. The brain sites involved in CRF's role in yohimbine-induced alcohol taking and seeking are unknown. We report that injections of the CRF receptor antagonist d-Phe CRF into the median raphe nucleus (MRN) attenuated yohimbine-induced reinstatement of alcohol seeking but had no effect on yohimbine-induced increases in alcohol intake during ongoing self-administration. Results indicate an important role of MRN CRF receptors in yohimbine-induced reinstatement of alcohol seeking but not yohimbine-induced increases in alcohol intake. © 2011 The Authors, Addiction Biology © 2011 Society for the Study of Addiction.

  20. Infralimbic and dorsal raphé microinjection of the 5-HT1B receptor agonist CP-93,129: attenuation of aggressive behavior in CFW male mice (United States)

    Faccidomo, S; Quadros, IMH; Takahashi, A; Fish, EW; Miczek, KA


    Rationale Aggressive behavior and impaired impulse control have been associated with dysregulations in the serotonergic system and with impaired functioning of the prefrontal cortex. 5-HT1B receptors have been shown to specifically modulate several types of offensive aggression. Objective To characterize the relative importance of 2 populations of 5-HT1B receptors in the dorsal raphé nucleus (DRN) and infralimbic cortex (ILC) in the modulation of aggressive behavior. Methods Male CFW mice were conditioned on a fixed-ratio 5 schedule of reinforcement to self-administer a 6% (w/v) alcohol solution. Mice repeatedly engaged in 5 min aggressive confrontations until aggressive behavior stabilized. Next, a cannula was implanted into either the DRN or the ILC. After recovery, mice were tested for aggression after self-administration of either 1.0 g/kg alcohol or water prior to a microinjection of the 5-HT1B agonist, CP-93,129 (0–1.0 µg/infusion). Results In both the DRN and ILC, CP-93,129 reduced aggressive behaviors after both water and alcohol self-administration. Intra-raphé CP-93,129 dose-dependently reduced both aggressive and locomotor behaviors. However, the anti-aggressive effects of intra-cortical CP-93,129 were behaviorally specific. Conclusions These findings highlight the importance of the serotonergic system in the modulation of aggression and suggest that the behaviorally specific effects of 5-HT1B receptor agonists are regionally selective. 5-HT1B receptors in a medial subregion of the prefrontal cortex, the ILC, appear to be critically involved in the attenuation of species-typical levels of aggression. PMID:22222863

  1. Bupropion-induced inhibition of α7 nicotinic acetylcholine receptors expressed in heterologous cells and neurons from dorsal raphe nucleus and hippocampus. (United States)

    Vázquez-Gómez, Elizabeth; Arias, Hugo R; Feuerbach, Dominik; Miranda-Morales, Marcela; Mihailescu, Stefan; Targowska-Duda, Katarzyna M; Jozwiak, Krzysztof; García-Colunga, Jesús


    The pharmacological activity of bupropion was compared between α7 nicotinic acetylcholine receptors expressed in heterologous cells and hippocampal and dorsal raphe nucleus neurons. The inhibitory activity of bupropion was studied on GH3-α7 cells by Ca2+ influx, as well as on neurons from the dorsal raphe nucleus and interneurons from the stratum radiatum of the hippocampal CA1 region by using a whole-cell voltage-clamp technique. In addition, the interaction of bupropion with the α7 nicotinic acetylcholine receptor was determined by [3H]imipramine competition binding assays and molecular docking. The fast component of acetylcholine- and choline-induced currents from both brain regions was inhibited by methyllycaconitine, indicating the participation of α7-containing nicotinic acetylcholine receptors. Choline-induced currents in hippocampal interneurons were partially inhibited by 10 µM bupropion, a concentration that could be reached in the brain during clinical administration. Additionally, both agonist-induced currents were reversibly inhibited by bupropion at concentrations that coincide with its inhibitory potency (IC50=54 µM) and binding affinity (Ki=63 µM) for α7 nicotinic acetylcholine receptors from heterologous cells. The [3H]imipramine competition binding and molecular docking results support a luminal location for the bupropion binding site(s). This study may help to understand the mechanisms of actions of bupropion at neuronal and molecular levels related with its therapeutic actions on depression and for smoking cessation. Copyright © 2014 Elsevier B.V. All rights reserved.

  2. Infralimbic and dorsal raphé microinjection of the 5-HT(1B) receptor agonist CP-93,129: attenuation of aggressive behavior in CFW male mice. (United States)

    Faccidomo, S; Quadros, I M H; Takahashi, A; Fish, E W; Miczek, K A


    Aggressive behavior and impaired impulse control have been associated with dysregulations in the serotonergic system and with impaired functioning of the prefrontal cortex. 5-HT(1B) receptors have been shown to specifically modulate several types of offensive aggression. This study aims to characterize the relative importance of two populations of 5-HT(1B) receptors in the dorsal raphé nucleus (DRN) and infralimbic cortex (ILC) in the modulation of aggressive behavior. Male CFW mice were conditioned on a fixed-ratio 5 schedule of reinforcement to self-administer a 6% (w/v) alcohol solution. Mice repeatedly engaged in 5-min aggressive confrontations until aggressive behavior stabilized. Next, a cannula was implanted into either the DRN or the ILC. After recovery, mice were tested for aggression after self-administration of either 1.0 g/kg alcohol or water prior to a microinjection of the 5-HT(1B) agonist, CP-93,129 (0-1.0 μg/infusion). In both the DRN and ILC, CP-93,129 reduced aggressive behaviors after both water and alcohol self-administration. Intra-raphé CP-93,129 dose-dependently reduced both aggressive and locomotor behaviors. However, the anti-aggressive effects of intra-cortical CP-93,129 were behaviorally specific. These findings highlight the importance of the serotonergic system in the modulation of aggression and suggest that the behaviorally specific effects of 5-HT(1B) receptor agonists are regionally selective. 5-HT(1B) receptors in a medial subregion of the prefrontal cortex, the ILC, appear to be critically involved in the attenuation of species-typical levels of aggression.

  3. Deep brain stimulation of the dorsal raphe inhibits avoidance and escape reactions and activates forebrain regions related to the modulation of anxiety/panic. (United States)

    Wscieklica, Tatiana; Silva, Mariana S C F; Lemes, Jéssica A; Melo-Thomas, Liana; Céspedes, Isabel C; Viana, Milena B


    One of the main neurochemical systems associated with anxiety/panic is the serotonergic system originating from the dorsal raphe nucleus (DR). Previous evidence suggests that the DR is composed of distinct subpopulations of neurons, both morphologically and functionally distinct. It seems that mainly the dorsal region of the DR (DRD) regulates anxiety-related reactions, while lateral wings DR (lwDR) serotonin (5-HT) neurons inhibit panic-related responses. In this study we used the technique of deep brain stimulation (DBS) to investigate the role played by the DRD and lwDR in defense. Male Wistar rats were submitted to high-frequency stimulation (100μA, 100Hz) in one of the two DR regions for 1h and immediately after tested in the avoidance or escape tasks of the elevated T-maze (ETM). In clinical terms, these responses have been related to generalized anxiety and panic disorder, respectively. After being submitted to the ETM, animals were placed in an open field for locomotor activity assessment. An additional group of rats was submitted to DBS of the DRD or the lwDR and used for quantification of c-Fos immunoreactive (Fos-ir) neurons in brain regions related to the modulation of defense. Results showed that stimulation of the DRD decreased avoidance latencies, an anxiolytic-like effect. DRD stimulation also led to increases in Fos-ir in the medial amygdala, lateral septum and cingulate cortex. DBS applied to the lwDR increased escape latencies, a panicolytic-like effect. This data highlights the importance of raphe topography and the potential benefit of the DBS technique for the treatment of anxiety-related disorders. Copyright © 2017 Elsevier B.V. All rights reserved.

  4. Create Your Plate

    Medline Plus

    Full Text Available ... Plate It's simple and effective for both managing diabetes and losing weight. Creating your plate lets you still choose the ... Help Enroll in the Living WIth Type 2 Diabetes Program Food & Fitness Food Recipes Planning Meals What Can I Eat Weight Loss Fitness In My Community Calendar of Events ...

  5. Create Your Plate

    Medline Plus

    Full Text Available ... For Lawyers Food & Fitness Home Food MyFoodAdvisor Recipes Association Cookbook Recipes Planning Meals Diabetes Meal Plans Create ... Become a Volunteer American Diabetes Month® American Diabetes Association Alert Day® Become a Member Advocacy Home Take ...

  6. Creating an Interactive PDF (United States)

    Branzburg, Jeffrey


    There are many ways to begin a PDF document using Adobe Acrobat. The easiest and most popular way is to create the document in another application (such as Microsoft Word) and then use the Adobe Acrobat software to convert it to a PDF. In this article, the author describes how he used Acrobat's many tools in his project--an interactive…

  7. Creating resilient SMEs

    DEFF Research Database (Denmark)

    Dahlberg, Rasmus; Guay, Fanny


    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...

  8. Create Your Plate

    Medline Plus

    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 ... your year-end donation today and help fight diabetes. Donate Today We Can Help - we-can-help. ...

  9. Create Your Plate

    Medline Plus

    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 ...

  10. Creating snags with explosives. (United States)

    Evelyn L. Bull; Arthur D. Partridge; Wayne G. Williams


    The tops of ponderosa pine (Pinus ponderosa) trees were blown off with dynamite to create nest sites for cavity-nesting wildlife. The procedure included drilling a hole almost through the trunk, inserting the dynamite, and setting the charge with primacord and fuse. Trees were simultaneously innoculated with a decay organism. The average cost was $...

  11. Creating Special Events (United States)

    deLisle, Lee


    "Creating Special Events" is organized as a systematic approach to festivals and events for students who seek a career in event management. This book looks at the evolution and history of festivals and events and proceeds to the nuts and bolts of event management. The book presents event management as the means of planning, organizing, directing,…

  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. Create Your Plate

    Medline Plus

    Full Text Available ... Your Plate Gluten Free Diets Meal Planning for Vegetarian Diets Cook with Heart-Healthy Foods Holiday Meal Planning What Can I Eat? ... Healthy Diet Create Your Plate Meal Planning for Vegetarian Diets Gluten Free Diets Holiday ... Foods donate en -- Limited Time MATCH Offer - limited- ...

  14. Creating a Virtual Gymnasium (United States)

    Fiorentino, Leah H.; Castelli, Darla


    Physical educators struggle with the challenges of assessing student performance, providing feedback about motor skills, and creating opportunities for all students to engage in game-play on a daily basis. The integration of technology in the gymnasium can address some of these challenges by improving teacher efficiency and increasing student…

  15. Creating Pupils' Internet Magazine (United States)

    Bognar, Branko; Šimic, Vesna


    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…

  16. Ordering of the N-terminus of human MDM2 by small molecule inhibitors. (United States)

    Michelsen, Klaus; Jordan, John B; Lewis, Jeffrey; Long, Alexander M; Yang, Evelyn; Rew, Yosup; Zhou, Jing; Yakowec, Peter; Schnier, Paul D; Huang, Xin; Poppe, Leszek


    Restoration of p53 function through the disruption of the MDM2-p53 protein complex is a promising strategy for the treatment of various types of cancer. Here, we present kinetic, thermodynamic, and structural rationale for the remarkable potency of a new class of MDM2 inhibitors, the piperidinones. While these compounds bind to the same site as previously reported for small molecule inhibitors, such as the Nutlins, data presented here demonstrate that the piperidinones also engage the N-terminal region (residues 10-16) of human MDM2, in particular, Val14 and Thr16. This portion of MDM2 is unstructured in both the apo form of the protein and in MDM2 complexes with p53 or Nutlin, but adopts a novel β-strand structure when complexed with the piperidinones. The ordering of the N-terminus upon binding of the piperidinones extends the current model of MDM2-p53 interaction and provides a new route to rational design of superior inhibitors.

  17. The N Terminus of α-Synuclein Forms Cu(II)-Bridged Oligomers. (United States)

    Drew, Simon C


    The oligomerization of α-synuclein (αSyn) is one of the defining features of Parkinson's disease. Binding of divalent copper to the N terminus of αSyn has been implicated in both its function and dysfunction. Herein, the molecular details of the Cu(II) /αSyn binding interface have been revealed using a library of synthetic 56-residue αSyn peptides containing site-specific isotopic labels. Using electron paramagnetic resonance spectroscopy, αSyn is shown to coordinate Cu(II) with high affinity via two pH-dependent coordination modes between pH 6.5-8.5. Most remarkably, the data demonstrate that the dominant mode is associated with binding to oligomers (antiparallel dimers and/or cyclic trimers) in which Cu(II) ions occupy intermolecular bridging sites. The findings provide a molecular link between Cu(II) -bound αSyn and its associated quaternary oligomeric structure. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  18. Creating bulk nanocrystalline metal.

    Energy Technology Data Exchange (ETDEWEB)

    Fredenburg, D. Anthony (Georgia Institute of Technology, Atlanta, GA); Saldana, Christopher J. (Purdue University, West Lafayette, IN); Gill, David D.; Hall, Aaron Christopher; Roemer, Timothy John (Ktech Corporation, Albuquerque, NM); Vogler, Tracy John; Yang, Pin


    Nanocrystalline and nanostructured materials offer unique microstructure-dependent properties that are superior to coarse-grained materials. These materials have been shown to have very high hardness, strength, and wear resistance. However, most current methods of producing nanostructured materials in weapons-relevant materials create powdered metal that must be consolidated into bulk form to be useful. Conventional consolidation methods are not appropriate due to the need to maintain the nanocrystalline structure. This research investigated new ways of creating nanocrystalline material, new methods of consolidating nanocrystalline material, and an analysis of these different methods of creation and consolidation to evaluate their applicability to mesoscale weapons applications where part features are often under 100 {micro}m wide and the material's microstructure must be very small to give homogeneous properties across the feature.

  19. Creating organizational cultures

    DEFF Research Database (Denmark)

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


    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...... is provided of the merits of this approach by revisiting the case of Enron. Originality/value – The paper constitutes an initial exploration of how social scientific and rhetorical perspectives on organizational change may be brought closer together. It may provide the first step towards the development...

  20. Can Computers Create Art?


    Hertzmann, Aaron


    This paper discusses whether computers, using Artifical Intelligence (AI), could create art. The first part concerns AI-based tools for assisting with art making. The history of technologies that automated aspects of art is covered, including photography and animation. In each case, we see initial fears and denial of the technology, followed by a blossoming of new creative and professional opportunities for artists. The hype and reality of Artificial Intelligence (AI) tools for art making is ...

  1. Creating product line architectures


    Bayer, J.; Flege, O.; Gacek, C.


    The creation and validation of product line software architectures are inherently more complex than those of software architectures for single systems. This paper compares a process for creating and evaluating a traditional, one-of-a- kind software architecture with one for a reference software architecture. The comparison is done in the context of PuLSE-DSSA, a customizable process that integrates both product line architecture creation and evaluation.

  2. Creating flat design websites

    CERN Document Server

    Pratas, Antonio


    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.

  3. Differential stress induced c-Fos expression and identification of region-specific miRNA-mRNA networks in the dorsal raphe and amygdala of high-responder/low-responder rats. (United States)

    Cohen, Joshua L; Ata, Anooshah E; Jackson, Nateka L; Rahn, Elizabeth J; Ramaker, Ryne C; Cooper, Sara; Kerman, Ilan A; Clinton, Sarah M


    Chronic stress triggers a variety of physical and mental health problems, and how individuals cope with stress influences risk for emotional disorders. To investigate molecular mechanisms underlying distinct stress coping styles, we utilized rats that were selectively-bred for differences in emotionality and stress reactivity. We show that high novelty responding (HR) rats readily bury a shock probe in the defensive burying test, a measure of proactive stress coping behavior, while low novelty responding (LR) rats exhibit enhanced immobility, a measure of reactive coping. Shock exposure in the defensive burying test elicited greater activation of HR rats' caudal dorsal raphe serotonergic cells compared to LRs, but lead to more pronounced activation throughout LRs' amygdala (lateral, basolateral, central, and basomedial nuclei) compared to HRs. RNA-sequencing revealed 271 mRNA transcripts and 33 microRNA species that were differentially expressed in HR/LR raphe and amygdala. We mapped potential microRNA-mRNA networks by correlating and clustering mRNA and microRNA expression and identified networks that differed in either the HR/LR dorsal raphe or amygdala. A dorsal raphe network linked three microRNAs which were down-regulated in LRs (miR-206-3p, miR-3559-5p, and miR-378a-3p) to repression of genes related to microglia and immune response (Cd74, Cyth4, Nckap1l, and Rac2), the genes themselves were up-regulated in LR dorsal raphe. In the amygdala, another network linked miR-124-5p, miR-146a-5p, miR-3068-3p, miR-380-5p, miR-539-3p, and miR-7a-1-3p with repression of chromatin remodeling-related genes (Cenpk, Cenpq, Itgb3bp, and Mis18a). Overall this work highlights potential drivers of gene-networks and downstream molecular pathways within the raphe and amygdala that contribute to individual differences in stress coping styles and stress vulnerabilities. Copyright © 2016 Elsevier B.V. All rights reserved.

  4. Creating Geoscience Leaders (United States)

    Buskop, J.; Buskop, W.


    The United Nations Educational, Scientific, and Cultural Organization recognizes 21 World Heritage in the United States, ten of which have astounding geological features: Wrangell St. Elias National Park, Olympic National Park, Mesa Verde National Park, Chaco Canyon, Glacier National Park, Carlsbad National Park, Mammoth Cave, Great Smokey Mountains National Park, Hawaii Volcanoes National Park, and Everglades National Park. Created by a student frustrated with fellow students addicted to smart phones with an extreme lack of interest in the geosciences, one student visited each World Heritage site in the United States and created one e-book chapter per park. Each chapter was created with original photographs, and a geological discovery hunt to encourage teen involvement in preserving remarkable geological sites. Each chapter describes at least one way young adults can get involved with the geosciences, such a cave geology, glaciology, hydrology, and volcanology. The e-book describes one park per chapter, each chapter providing a geological discovery hunt, information on how to get involved with conservation of the parks, geological maps of the parks, parallels between archaeological and geological sites, and how to talk to a ranger. The young author is approaching UNESCO to publish the work as a free e-book to encourage involvement in UNESCO sites and to prove that the geosciences are fun.

  5. Der terminus προνοητησ in der Byzantinischen verwaltung

    Directory of Open Access Journals (Sweden)

    Wassiliou-Seibt Alexandra-Kyriaki


    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.

  6. Computational analysis of the CB1 carboxyl-terminus in the receptor-G protein complex. (United States)

    Shim, Joong-Youn; Khurana, Leepakshi; Kendall, Debra A


    Despite the important role of the carboxyl-terminus (Ct) of the activated brain cannabinoid receptor one (CB1) in the regulation of G protein signaling, a structural understanding of interactions with G proteins is lacking. This is largely due to the highly flexible nature of the CB1 Ct that dynamically adapts its conformation to the presence of G proteins. In the present study, we explored how the CB1 Ct can interact with the G protein by building on our prior modeling of the CB1-Gi complex (Shim, Ahn, and Kendall, The Journal of Biological Chemistry 2013;288:32449-32465) to incorporate a complete CB1 Ct (Glu416(Ct)-Leu472(Ct)). Based on the structural constraints from NMR studies, we employed ROSETTA to predict tertiary folds, ZDOCK to predict docking orientation, and molecular dynamics (MD) simulations to obtain two distinct plausible models of CB1 Ct in the CB1-Gi complex. The resulting models were consistent with the NMR-determined helical structure (H9) in the middle region of the CB1 Ct. The CB1 Ct directly interacted with both Gα and Gβ and stabilized the receptor at the Gi interface. The results of site-directed mutagenesis studies of Glu416(Ct), Asp423(Ct), Asp428(Ct), and Arg444(Ct) of CB1 Ct suggested that the CB1 Ct can influence receptor-G protein coupling by stabilizing the receptor at the Gi interface. This research provided, for the first time, models of the CB1 Ct in contact with the G protein. © 2016 Wiley Periodicals, Inc.

  7. Antibodies against the C-terminus of α-synuclein modulate its fibrillation. (United States)

    Sahin, Cagla; Lorenzen, Nikolai; Lemminger, Lasse; Christiansen, Gunna; Møller, Ian Max; Vesterager, Louise Buur; Pedersen, Lars Østergaard; Fog, Karina; Kallunki, Pekka; Otzen, Daniel E


    The 140-residue natively disordered protein α-synuclein (aSN) is a central component in the development of a family of neurodegenerative diseases termed synucleinopathies. This is attributed to its ability to form cytotoxic aggregates such as oligomers and amyloid fibrils. Consequently there have been intense efforts to avoid aggregation or reroute the aggregation pathway using pharmaceutical agents such as small molecules, chaperones and antibodies. aSN's lack of persistent structure in the monomeric state, as well as the multitude of different oligomeric and even different fibrillar states, makes it difficult to raise antibodies that would be efficacious in neutralizing all conformations of aSN. However, the C-terminal 20-40 residues of aSN are a promising epitope for antibody development. It is primarily disordered in both monomeric and aggregated forms, and an anti-C-terminal antibody will therefore be able to bind all forms. Furthermore, it might not interfere with the folding of aSN into membranes, which could be important for its physiological role. Here we report a screen of a series of monoclonal antibodies, which all target the C-terminal of aSN. According to dot blot analyses, different antibodies bound different forms of aSN with different preferences and showed reduced binding to monomeric compared to aggregated (oligomeric and fibrillary) aSN. Consequently they have different effects on aSN's ability to fibrillate and permeabilize membranes. Generally, the antibodies with strongest binding to aggregated aSN in dot blot, also inhibited fibrillation and membrane permeabilization the most, and promoted formation of amorphous aggregates surrounded by small and thin fibers. This suggests that the development of antibodies that targets the C-terminus, exposed in the aggregated forms of aSN, may be beneficial for improved immunotherapy against PD. Copyright © 2016 Elsevier B.V. All rights reserved.

  8. The NH2terminus regulates voltage-dependent gating of CALHM ion channels. (United States)

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


    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.

  9. A Peptide with a ProGln C Terminus in the Human Saliva Peptidome Exerts Bactericidal Activity against Propionibacterium acnes▿ (United States)

    Huang, Chun-Ming; Torpey, Justin W.; Liu, Yu-Tseung; Chen, Yun-Ru; Williams, Katherine E.; Komives, Elizabeth A.; Gallo, Richard L.


    Nine proline-rich peptides ending with a proline-glutamine C terminus in a salivary peptidome were sequenced by matrix-assisted laser desorption ionization time of flight time of flight tandem mass spectrometry. A GPPPQGGRPQ peptide binds gram-positive Propionibacterium acnes and considerably inhibits bacterial growth. The peptide exhibiting innate immunity may be applied for treatment of various P. acnes-associated human diseases. PMID:18285475

  10. Functional and biophysical analysis of the C-terminus of the CGRP-receptor; a family B GPCR. (United States)

    Conner, Matthew; Hicks, Matthew R; Dafforn, Tim; Knowles, Timothy J; Ludwig, Christian; Staddon, Susan; Overduin, Michael; Günther, Ulrich L; Thome, Johannes; Wheatley, Mark; Poyner, David R; Conner, Alex C


    G-protein coupled receptors (GPCRs) typically have a functionally important C-terminus which, in the largest subfamily (family A), includes a membrane-parallel eighth helix. Mutations of this region are associated with several diseases. There are few C-terminal studies on the family B GPCRs and no data supporting the existence of a similar eighth helix in this second major subfamily, which has little or no sequence homology to family A GPCRs. Here we show that the C-terminus of a family B GPCR (CLR) has a disparate region from N400 to C436 required for CGRP-mediated internalization, and a proximal region of twelve residues (from G388 to W399), in a similar position to the family A eighth helix, required for receptor localization at the cell surface. A combination of circular and linear dichroism, fluorescence and modified waterLOGSY NMR spectroscopy (SALMON) demonstrated that a peptide mimetic of this domain readily forms a membrane-parallel helix anchored to the liposome by an interfacial tryptophan residue. The study reveals two key functions held within the C-terminus of a family B GPCR and presents support for an eighth helical region with striking topological similarity to the nonhomologous family A receptor. This helix structure appears to be found in most other family B GPCRs.

  11. Calmodulin Kinase II Interacts with the Dopamine Transporter C Terminus to Regulate Amphetamine-Induced Reverse Transport

    DEFF Research Database (Denmark)

    Fog, Jacob U; Khoshbouei, Habibeh; Holy, Marion


    Efflux of dopamine through the dopamine transporter (DAT) is critical for the psychostimulatory properties of amphetamines, but the underlying mechanism is unclear. Here we show that Ca(2+)/calmodulin-dependent protein kinase II (CaMKII) plays a key role in this efflux. CaMKIIalpha bound to the d......Efflux of dopamine through the dopamine transporter (DAT) is critical for the psychostimulatory properties of amphetamines, but the underlying mechanism is unclear. Here we show that Ca(2+)/calmodulin-dependent protein kinase II (CaMKII) plays a key role in this efflux. CaMKIIalpha bound...... to the distal C terminus of DAT and colocalized with DAT in dopaminergic neurons. CaMKIIalpha stimulated dopamine efflux via DAT in response to amphetamine in heterologous cells and in dopaminergic neurons. CaMKIIalpha phosphorylated serines in the distal N terminus of DAT in vitro, and mutation...... of these serines eliminated the stimulatory effects of CaMKIIalpha. A mutation of the DAT C terminus impairing CaMKIIalpha binding also impaired amphetamine-induced dopamine efflux. An in vivo role for CaMKII was supported by chronoamperometry measurements showing reduced amphetamine-induced dopamine efflux...

  12. Calmodulin kinase II interacts with the dopamine transporter C terminus to regulate amphetamine-induced reverse transport

    DEFF Research Database (Denmark)

    Fog, Jacob U; Khoshbouei, Habibeh; Holy, Marion


    Efflux of dopamine through the dopamine transporter (DAT) is critical for the psychostimulatory properties of amphetamines, but the underlying mechanism is unclear. Here we show that Ca(2+)/calmodulin-dependent protein kinase II (CaMKII) plays a key role in this efflux. CaMKIIalpha bound to the d......Efflux of dopamine through the dopamine transporter (DAT) is critical for the psychostimulatory properties of amphetamines, but the underlying mechanism is unclear. Here we show that Ca(2+)/calmodulin-dependent protein kinase II (CaMKII) plays a key role in this efflux. CaMKIIalpha bound...... to the distal C terminus of DAT and colocalized with DAT in dopaminergic neurons. CaMKIIalpha stimulated dopamine efflux via DAT in response to amphetamine in heterologous cells and in dopaminergic neurons. CaMKIIalpha phosphorylated serines in the distal N terminus of DAT in vitro, and mutation...... of these serines eliminated the stimulatory effects of CaMKIIalpha. A mutation of the DAT C terminus impairing CaMKIIalpha binding also impaired amphetamine-induced dopamine efflux. An in vivo role for CaMKII was supported by chronoamperometry measurements showing reduced amphetamine-induced dopamine efflux...

  13. Creating sustainable performance. (United States)

    Spreitzer, Gretchen; Porath, Christine


    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.

  14. Tourist-created Content

    DEFF Research Database (Denmark)

    Munar, Ana Maria


    study of social media sites and destination brands, relying on qualitative research methods, content analysis and field research. Findings – Tourists are largely contributing to destination image formation, while avoiding the use of the formal elements of the brands. The most popular strategies used...... by destination management organizations exhibit some crucial weaknesses. However, a strategy based on analytics brings new opportunities for destination branding. Originality/value – The study provides an innovative analysis of tourist-created content and its impact on destination branding and presents......Purpose – The purpose of this paper is to analyze the relationship between tourists' user-generated content on the web and destination branding, as well as to discuss the online strategies used by destination management organizations. Design/methodology/approach – The research adopts an exploratory...

  15. Creating nanostars with buckyballs

    Energy Technology Data Exchange (ETDEWEB)

    Bae, Young K., E-mail:


    We report creating superradiant quantum nanoplasmas (nanostars) by impacting buckyballs at hypervelocities (v>100 km/s) in an innovative tabletop apparatus. The nanostars are estimated to have ∼10 TPa transient pressures and convert ∼35% of impact energy into soft-X-ray energy. The ultrahigh-efficiency conversion is proposed to result from Dicke Superradiance of Metastable Innershell Molecular State, originally discovered by the author and his colleagues in 1994. The usage of buckyballs and successful orders-of-magnitude scaling down of the apparatus size and complexity establish an innovative tabletop method for generating, studying, and utilizing matter in planetary or stellar interiors and open doors to numerous unprecedented applications.

  16. Creating Griffith Observatory (United States)

    Cook, Anthony


    Griffith Observatory has been the iconic symbol of the sky for southern California since it began its public mission on May 15, 1935. While the Observatory is widely known as being the gift of Col. Griffith J. Griffith (1850-1919), the story of how Griffith’s gift became reality involves many of the people better known for other contributions that made Los Angeles area an important center of astrophysics in the 20th century. Griffith began drawing up his plans for an observatory and science museum for the people of Los Angeles after looking at Saturn through the newly completed 60-inch reflector on Mt. Wilson. He realized the social impact that viewing the heavens could have if made freely available, and discussing the idea of a public observatory with Mt. Wilson Observatory’s founder, George Ellery Hale, and Director, Walter Adams. This resulted, in 1916, in a will specifying many of the features of Griffith Observatory, and establishing a committee managed trust fund to build it. Astronomy popularizer Mars Baumgardt convinced the committee at the Zeiss Planetarium projector would be appropriate for Griffith’s project after the planetarium was introduced in Germany in 1923. In 1930, the trust committee judged funds to be sufficient to start work on creating Griffith Observatory, and letters from the Committee requesting help in realizing the project were sent to Hale, Adams, Robert Millikan, and other area experts then engaged in creating the 200-inch telescope eventually destined for Palomar Mountain. A Scientific Advisory Committee, headed by Millikan, recommended that Caltech Physicist Edward Kurth be put in charge of building and exhibit design. Kurth, in turn, sought help from artist Russell Porter. The architecture firm of John C. Austin and Fredrick Ashley was selected to design the project, and they adopted the designs of Porter and Kurth. Philip Fox of the Adler Planetarium was enlisted to manage the completion of the Observatory and become its

  17. Creating the living brand. (United States)

    Bendapudi, Neeli; Bendapudi, Venkat


    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.

  18. LQT1 mutations in KCNQ1 C-terminus assembly domain suppress IKs using different mechanisms. (United States)

    Aromolaran, Ademuyiwa S; Subramanyam, Prakash; Chang, Donald D; Kobertz, William R; Colecraft, Henry M


    Long QT syndrome 1 (LQT1) mutations in KCNQ1 that decrease cardiac IKs (slowly activating delayed rectifier K(+) current) underlie ventricular arrhythmias and sudden death. LQT1 mutations may suppress IKs by preventing KCNQ1 assembly, disrupting surface trafficking, or inhibiting gating. We investigated mechanisms underlying how three LQT1 mutations in KCNQ1 C-terminus assembly domain (R555H/G589D/L619M) decrease IKs in heterologous cells and cardiomyocytes. In Chinese hamster ovary (CHO) cells, mutant KCNQ1 + KCNE1 channels either produced no currents (G589D/L619M) or displayed markedly reduced IKs with a right-shifted voltage-dependence of activation (R555H). When co-expressed with wild-type (wt) KCNQ1, the mutant KCNQ1s displayed varying intrinsic dominant-negative capacities that were affected by auxiliary KCNE1. All three mutant KCNQ1s assembled with wt KCNQ1 as determined by fluorescence resonance energy transfer (FRET). We developed an optical quantum dot labelling assay to measure channel surface density. G589D/R555H displayed substantial reductions in surface density, which were either partially (G589D) or fully (R555H) rescued by wt KCNQ1. Unexpectedly, L619M showed no trafficking defect. In adult rat cardiomyocytes, adenovirus-expressed homotetrameric G589D/L619M + KCNE1 channels yielded no currents, whereas R555H + KCNE1 produced diminished IKs with a right-shifted voltage-dependence of activation, mimicking observations in CHO cells. In contrast to heterologous cells, homotetrameric R555H channels showed no trafficking defect in cardiomyocytes. Distinct LQT1 mutations in KCNQ1 assembly domain decrease IKs using unique combinations of biophysical and trafficking mechanisms. Functional deficits in IKs observed in heterologous cells are mostly, but not completely, recapitulated in adult rat cardiomyocytes. A 'methodological chain' combining approaches in heterologous cells and cardiomyocytes provides mechanistic insights that may help advance personalized

  19. Creating alternatives in science (United States)


    Traditional scientist training at the PhD level does not prepare students to be competitive in biotechnology or other non-academic science careers. Some universities have developed biotechnology-relevant doctoral programmes, but most have not. Forming a life science career club makes a statement to university administrators that it is time to rework the curriculum to include biotechnology-relevant training. A career club can supplement traditional PhD training by introducing students to available career choices, help them develop a personal network and teach the business skills that they will need to be competitive in science outside of academia. This paper is an instructional guide designed to help students create a science career club at their own university. These suggestions are based on the experience gained in establishing such a club for the Graduate School at the University of Colorado Denver. We describe the activities that can be offered, the job descriptions for the offices required and potential challenges. With determination, a creative spirit, and the guidance of this paper, students should be able to greatly increase awareness of science career options, and begin building the skills necessary to become competitive in non-academic science. PMID:20161069

  20. Creating an open mind. (United States)

    Monaghan, Duncan


    Duncan Monaghan is 33 years old and in his second year of an Arts degree in Creative Writing. He is a published poet and is currently producing a music CD. Duncan has a history of bipolar disorder which was diagnosed when he was nineteen: "It worried me at first a lot. It played on my mind constantly. I felt different from everybody else--I did not understand what was happening to me." Drawing on his life experiences, Duncan has been enhancing his recovery through creativity--in poetry, lyrics, music and story. "Life for me was a constant battle of relying on medication and appointments with my case manager...until I realized I could combine my recovery with my passions as a tool to use as an outlet to many of the "mind traps" I so often found hindering my own recovery." Duncan is Aboriginal and has experience of the mental health systems in most states and territories and now lives in Brisbane. This is a shortened version of his presentation at Creating Futures 2010.

  1. Analysis of serotonin, dopamine and their metabolites in the caudate putamen, the suprachiasmatic nucleus and the median raphe nucleus of euthermic and torpid deermice, Peromyscus maniculatus. (United States)

    Lin, L H; Pivorun, E B


    Deermice, subjected to food rationing and low ambient temperature, were sacrificed in normothermia or during daily torpor. Levels of monoamines and their metabolites in the caudate putamen (CPN), the suprachiasmatic nuclear area (SCN), and the median raphe nucleus (MRN) were quantified through the use of HPLC with electrochemical detection. Significant elevations in levels (pg/mg protein) of the serotonin (5-HT) metabolite, 5-hydroxyindole-3-acetic acid (5-HIAA) were noted in torpid individuals in all nuclei examined. The dopamine (DA) metabolite, homovanillic acid (HVA) was significantly elevated in the CPN and MRN of torpid individuals. Moreover, a significant increase in the HVA to DA ratio was also noted in the CPN and the MRN. In the SCN, the concentrations of 5-hydroxytryptophan (5-HTP), 5-HT, DA and 3,4-dihydroxyphenylacetic acid (DOPAC) were also increased significantly during torpor. These significant elevations suggest that an increase in the activity of the serotonergic and dopaminergic systems occurs in these nuclei during daily torpor in the deermouse.

  2. The vigilance promoting drug modafinil modulates serotonin transmission in the rat prefrontal cortex and dorsal raphe nucleus. Possible relevance for its postulated antidepressant activity. (United States)

    Ferraro, Luca; Antonelli, Tiziana; Beggiato, Sarah; Cristina Tomasini, Maria; Fuxe, Kjell; Tanganelli, Sergio


    Modafinil, (RS)-2-(diphenylmethylsulfinyl)acetamide derivative (Modiodal, Provigil), is a vigilance-promoting agent which reduces sleep episodes by improving wakefulness. It is approved by the USA FDA for narcolepsy, shiftwork sleep disorder and obstructive sleep apnoea with residual excessive sleepiness despite optimal use of continuous positive airway pressure. Unlike classical psychostimulants such as amphetamine and amphetamine-like compounds, the awaking effect of modafinil is not associated with a disturbance of nighttime sleep, tolerance, and sensitization. Its precise mechanism of action is still unclear. In animal studies, modafinil and its analogues have been shown to modify dopaminergic, noradrenergic, glutamatergic, GABAergic, serotoninergic, orexinergic, and histaminergic pathways. Besides the approved use in sleep disorders, modafinil has been investigated for the treatment of fatigue, impaired cognition and some symptoms in a number of other disorders. In particular, clinical studies seem to indicate that the drug could be particularly successful in the treatment of depression and its use in major depressive and bipolar disorders, has been suggested. However, the molecular mechanisms underlying this possible effect are still unknown. The present review firstly summarizes the structure-activity relationship studies and the mechanism of action of modafinil and its related compounds. Then, it focuses on data demonstrating that modafinil interacts with serotonin neuronal activity in rat frontal cortex and dorsal raphe nucleus, two brain areas linked together and involved in depression. Preclinical and clinical evidence of a positive interaction between modafinil and classical antidepressant drugs, is also summarized.

  3. How does early maternal separation and chronic stress in adult rats affect the immunoreactivity of serotonergic neurons within the dorsal raphe nucleus? (United States)

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


    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.

  4. C-terminus of a hexapeptidic ghrelin receptor inverse agonist can switch peptide behavior from inverse agonism to agonism. (United States)

    Els-Heindl, Sylvia; Bellmann-Sickert, Kathrin; Beck-Sickinger, Annette G


    Subtle changes in the sequence at the N-terminus and in the aromatic core of hexapeptidic ghrelin receptor inverse agonists can switch behavior from inverse agonism to agonism, but the C-terminal role of the sequence is unclear. Thus, analogs of the ghrelin receptor inverse agonist KbFwLL-NH2 (b = β-(3-benzothienyl)-d-alanine) were synthesized by solid phase peptide synthesis in order to identify the influence of aromaticity, charge, and hydrophobicity. Potency and efficacy of the hexapeptides were evaluated in inositol triphosphate turnover assays. Notably, modifications directly at the C-terminal Leu(6) could influence peptide efficacy leading to decreased constitutive activity. High hydrophobicity at the C-terminal position was of importance for elevated inverse agonist activity, the introduction of charged amino acids led to decreased potency. In contrast, structure-activity relationship studies of Leu(5) located closer to the aromatic core revealed an agonism-inducing position. These findings imply that amino acids with possible cation-π or π-π interactions and a suitable orientation at the C-terminus of the aromatic core induce agonism. Receptor binding studies showed that most peptides bind to the receptor at a concentration of 1 µM and modification directly at the C-terminus is generally more accepted than Leu(5) substitution. Interestingly, this observation is not dependent on the type of modification. These studies reveal another switch region of the short ghrelin receptor ligand pointing out the sensitivity of the ghrelin receptor binding pocket. © 2015 Wiley Periodicals, Inc.

  5. Pannexin-1 is blocked by its C-terminus through a delocalized non-specific interaction surface.

    Directory of Open Access Journals (Sweden)

    Michelle Dourado

    Full Text Available The Pannexin-1 (Panx1 channel is known to become activated under a variety of physiological conditions resulting in the release of medium-sized molecules such as ATP and amino acids from the cell. The detailed molecular mechanism of activation of the channel resulting in the opening of the Pannexin pore is poorly understood. The best-studied gating mechanism is caspase-3/7-mediated cleavage and truncation of the c-terminus. In the absence of caspase-cleavage, the c-terminal peptide maintains the channel in the closed state, possibly by directly plugging the pore from the intracellular side. We sought to understand in detail the part of the c-terminus necessary for this interaction by alanine-scanning and truncation mutagenesis of the c-terminal gating peptide. These experiments demonstrate that no single amino acid side-chain is necessary for this interaction. In fact, replacing blocks of 10-12 amino acids in different parts of the c-terminal peptide with alanines fails to disrupt the ability of the c-terminus to keep the channel closed. Surprisingly, even replacing the entire c-terminal gating peptide with a scrambled peptide of the same length maintains the interaction in some cases. Further analysis revealed that the interaction surface, while delocalized, is located within the amino-terminal two-thirds of the c-terminal peptide. Such a delocalized and potentially low-affinity interaction surface is allowed due to the high effective concentration of the c-terminal peptide near the inner vestibule of the pore and likely explains why this region is poorly conserved between species. This type of weak interaction with a tethered gating peptide may be required to maintain high-sensitivity to caspase-dependent activation.

  6. Autographa californica multiple nucleopolyhedrovirus DNA polymerase C terminus is required for nuclear localization and viral DNA replication. (United States)

    Feng, Guozhong; Krell, Peter J


    The DNA polymerase (DNApol) of the baculovirus Autographa californica multiple nucleopolyhedrovirus (AcMNPV) is essential for viral DNA replication. The DNApol exonuclease and polymerase domains are highly conserved and are considered functional in DNA replication. However, the role of the DNApol C terminus has not yet been characterized. To identify whether only the exonuclease and polymerase domains are sufficient for viral DNA replication, several DNApol C-terminal truncations were cloned into a dnapol-null AcMNPV bacmid with a green fluorescent protein (GFP) reporter. Surprisingly, most of the truncation constructs, despite containing both exonuclease and polymerase domains, could not rescue viral DNA replication and viral production in bacmid-transfected Sf21 cells. Moreover, GFP fusions of these same truncations failed to localize to the nucleus. Truncation of the C-terminal amino acids 950 to 984 showed nuclear localization but allowed for only limited and delayed viral spread. The C terminus contains a typical bipartite nuclear localization signal (NLS) motif at residues 804 to 827 and a monopartite NLS motif at residues 939 to 948. Each NLS, as a GFP fusion peptide, localized to the nucleus, but both NLSs were required for nuclear localization of DNApol. Alanine substitutions in a highly conserved baculovirus DNApol sequence at AcMNPV DNApol amino acids 972 to 981 demonstrated its importance for virus production and DNA replication. Collectively, the data indicated that the C terminus of AcMNPV DNApol contains two NLSs and a conserved motif, all of which are required for nuclear localization of DNApol, viral DNA synthesis, and virus production. The baculovirus DNA polymerase (DNApol) is a highly specific polymerase that allows viral DNA synthesis and hence virus replication in infected insect cells. We demonstrated that the exonuclease and polymerase domains of Autographa californica multiple nucleopolyhedrovirus (AcMNPV) alone are insufficient for viral

  7. Aminoacylase 3 binds to and cleaves the N-terminus of the hepatitis C virus core protein (United States)

    Tsirulnikov, Kirill; Abuladze, Natalia; Vahi, Ritu; Hasnain, Huma; Phillips, Martin; Ryan, Christopher M.; Atanasov, Ivo; Faull, Kym F.; Kurtz, Ira; Pushkin, Alexander


    Aminoacylase 3 (AA3) mediates deacetylation of N-acetyl aromatic amino acids and mercapturic acids. Deacetylation of mercapturic acids of exo- and endobiotics are likely involved in their toxicity. AA3 is predominantly expressed in kidney, and to a lesser extent in liver, brain, and blood. AA3 has been recently reported to interact with the hepatitis C virus core protein (HCVCP) in the yeast two-hybrid system. Here we demonstrate that AA3 directly binds to HCVCP (Kd~10 μM) that may by implicated in HCV pathogenesis. AA3 also revealed a weak endopeptidase activity towards the N-terminus of HCVCP. PMID:23010594

  8. Pathological conformations involving the amino terminus of tau occur early in Alzheimer's disease and are differentially detected by monoclonal antibodies. (United States)

    Combs, Benjamin; Hamel, Chelsey; Kanaan, Nicholas M


    Conformational changes involving the amino terminus of the tau protein are among the earliest alterations associated with tau pathology in Alzheimer's disease and other tauopathies. This region of tau contains a phosphatase-activating domain (PAD) that is aberrantly exposed in pathological forms of the protein, an event that is associated with disruptions in anterograde fast axonal transport. We utilized four antibodies that recognize the amino terminus of tau, TNT1, TNT2 (a novel antibody), Tau12, and Tau13, to further study this important region. Using scanning alanine mutations in recombinant tau proteins, we refined the epitopes of each antibody. We examined the antibodies' relative abilities to specifically label pathological tau in non-denaturing and denaturing assays to gain insight into some of the mechanistic details of PAD exposure. We then determined the pattern of tau pathology labeled by each antibody in human hippocampal sections at various disease stages in order to characterize PAD exposure in the context of disease progression. The characteristics of reactivity for the antibodies fell into two groups. TNT1 and TNT2 recognized epitopes within amino acids 7-12 and specifically identified recombinant tau aggregates and pathological tau from Alzheimer's disease brains in a conformation-dependent manner. These antibodies labeled early pre-tangle pathology from neurons in early Braak stages and colocalized with thiazine red, a marker of fibrillar pathology, in classic neurofibrillary tangles. However, late tangles were negative for TNT1 and TNT2 indicating a loss of the epitope in later stages of tangle evolution. In contrast, Tau12 and Tau13 both identified discontinuous epitopes in the amino terminus and were unable to differentiate between normal and pathological tau in biochemical and tissue immunohistological assays. Despite the close proximity of these epitopes, the antibodies demonstrated remarkably different abilities to identify pathological

  9. 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† (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.


    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

  10. The C-terminus of PufX plays a key role in dimerisation and assembly of the reaction center light-harvesting 1 complex from Rhodobacter sphaeroides. (United States)

    Qian, Pu; Martin, Elizabeth C; Ng, Irene W; Hunter, C Neil


    In bacterial photosynthesis reaction center-light-harvesting 1 (RC-LH1) complexes trap absorbed solar energy by generating a charge separated state. Subsequent electron and proton transfers form a quinol, destined to diffuse to the cytochrome bc1 complex. In bacteria such as Rhodobacter (Rba.) sphaeroides and Rba. capsulatus the PufX polypeptide creates a channel for quinone/quinol traffic across the LH1 complex that surrounds the RC, and it is therefore essential for photosynthetic growth. PufX also plays a key role in dimerization of the RC-LH1-PufX core complex, and the structure of the Rba. sphaeroides complex shows that the PufX C-terminus, particularly the region from X49-X53, likely mediates association of core monomers. To investigate this putative interaction we analysed mutations PufX R49L, PufX R53L, PufX R49/53L and PufX G52L by measuring photosynthetic growth, fractionation of detergent-solubilised membranes, formation of 2-D crystals and electron microscopy. We show that these mutations do not affect assembly of PufX within the core or photosynthetic growth but they do prevent dimerization, consistent with predictions from the RC-LH1-PufX structure. We obtained low resolution structures of monomeric core complexes with and without PufX, using electron microscopy of negatively stained single particles and 3D reconstruction; the monomeric complex with PufX corresponds to one half of the dimer structure whereas LH1 completely encloses the RC if the gene encoding PufX is deleted. On the basis of the insights gained from these mutagenesis and structural analyses we propose a sequence for assembly of the dimeric RC-LH1-PufX complex. Crown Copyright © 2017. Published by Elsevier B.V. All rights reserved.

  11. A functional dissection of PTEN N-terminus: implications in PTEN subcellular targeting and tumor suppressor activity. (United States)

    Gil, Anabel; Rodríguez-Escudero, Isabel; Stumpf, Miriam; Molina, María; Cid, Víctor J; Pulido, Rafael


    Spatial regulation of the tumor suppressor PTEN is exerted through alternative plasma membrane, cytoplasmic, and nuclear subcellular locations. The N-terminal region of PTEN is important for the control of PTEN subcellular localization and function. It contains both an active nuclear localization signal (NLS) and an overlapping PIP2-binding motif (PBM) involved in plasma membrane targeting. We report a comprehensive mutational and functional analysis of the PTEN N-terminus, including a panel of tumor-related mutations at this region. Nuclear/cytoplasmic partitioning in mammalian cells and PIP3 phosphatase assays in reconstituted S. cerevisiae defined categories of PTEN N-terminal mutations with distinct PIP3 phosphatase and nuclear accumulation properties. Noticeably, most tumor-related mutations that lost PIP3 phosphatase activity also displayed impaired nuclear localization. Cell proliferation and soft-agar colony formation analysis in mammalian cells of mutations with distinctive nuclear accumulation and catalytic activity patterns suggested a contribution of both properties to PTEN tumor suppressor activity. Our functional dissection of the PTEN N-terminus provides the basis for a systematic analysis of tumor-related and experimentally engineered PTEN mutations.

  12. Absence of PTHrP nuclear localization and carboxyl terminus sequences leads to abnormal brain development and function.

    Directory of Open Access Journals (Sweden)

    Zhen Gu

    Full Text Available We assessed whether the nuclear localization sequences (NLS and C terminus of parathyroid hormone-related protein (PTHrP play critical roles in brain development and function. We used histology, immunohistochemistry, histomorphometry, Western blots and electrophysiological recordings to compare the proliferation and differentiation of neural stem cells, neuronal hippocampal synaptic transmission, and brain phenotypes including shape and structures, in Pthrp knock-in mice, which express PTHrP (1-84, a truncated form of the protein that is missing the NLS and the C-terminal region of the protein, and their wild-type littermates. Results showed that Pthrp knock-in mice display abnormal brain shape and structures; decreased neural cell proliferative capacity and increased apoptosis associated with up-regulation of cyclin dependent kinase inhibitors p16, p21, p27 and p53 and down-regulation of the Bmi-1 oncogene; delayed neural cell differentiation; and impaired hippocampal synaptic transmission and plasticity. These findings provide in vivo experimental evidence that the NLS and C-terminus of PTHrP are essential not only for the regulation of neural cell proliferation and differentiation, but also for the maintenance of normal neuronal synaptic transmission and plasticity.

  13. Glucagon-Like Peptide 1 and Its Analogs Act in the Dorsal Raphe and Modulate Central Serotonin to Reduce Appetite and Body Weight. (United States)

    Anderberg, Rozita H; Richard, Jennifer E; Eerola, Kim; López-Ferreras, Lorena; Banke, Elin; Hansson, Caroline; Nissbrandt, Hans; Berqquist, Filip; Gribble, Fiona M; Reimann, Frank; Wernstedt Asterholm, Ingrid; Lamy, Christophe M; Skibicka, Karolina P


    Glucagon-like peptide 1 (GLP-1) and serotonin play critical roles in energy balance regulation. Both systems are exploited clinically as antiobesity strategies. Surprisingly, whether they interact in order to regulate energy balance is poorly understood. Here we investigated mechanisms by which GLP-1 and serotonin interact at the level of the central nervous system. Serotonin depletion impaired the ability of exendin-4, a clinically used GLP-1 analog, to reduce body weight in rats, suggesting that serotonin is a critical mediator of the energy balance impact of GLP-1 receptor (GLP-1R) activation. Serotonin turnover and expression of 5-hydroxytryptamine (5-HT) 2A (5-HT2A) and 5-HT2C serotonin receptors in the hypothalamus were altered by GLP-1R activation. We demonstrate that the 5-HT2A, but surprisingly not the 5-HT2C, receptor is critical for weight loss, anorexia, and fat mass reduction induced by central GLP-1R activation. Importantly, central 5-HT2A receptors are also required for peripherally injected liraglutide to reduce feeding and weight. Dorsal raphe (DR) harbors cell bodies of serotonin-producing neurons that supply serotonin to the hypothalamic nuclei. We show that GLP-1R stimulation in DR is sufficient to induce hypophagia and increase the electrical activity of the DR serotonin neurons. Finally, our results disassociate brain metabolic and emotionality pathways impacted by GLP-1R activation. This study identifies serotonin as a new critical neural substrate for GLP-1 impact on energy homeostasis and expands the current map of brain areas impacted by GLP-1R activation. © 2017 by the American Diabetes Association.

  14. Chronic social defeat up-regulates expression of the serotonin transporter in rat dorsal raphe nucleus and projection regions in a glucocorticoid-dependent manner. (United States)

    Zhang, Jia; Fan, Yan; Li, Ying; Zhu, Hobart; Wang, Liang; Zhu, Meng-Yang


    Chronic stress and dysfunction of the serotonergic system in the brain have been considered two of the major risks for development of depression. In this study, adult Fischer 344 rats were subjected to a regimen of chronic social defeat (CSD). To mimic stressful conditions, some rats were not exposed to CSD, but instead treated with corticosterone (CORT) in oral solution while maintained in their home cage. Protein levels of the serotonin transporter (SERT) in the dorsal raphe nucleus (DRN), hippocampus, frontal cortex, and amygdala were examined by Western blotting or immunofluorescence staining. The results showed that CSD up-regulated SERT protein levels in the DRN, hippocampus, frontal cortex, and amygdala regions. This up-regulation was abolished or prevented by adrenalectomy, or treatment with antagonists of corticosteroid receptors mifepristone and spironolactone, alone or in combination. Similarly, up-regulated SERT protein levels in these brain regions were also observed in rats treated with oral CORT ingestion, which was analogously prevented by treatment with mifepristone and spironolactone. Furthermore, both CSD- and CORT-induced up-regulation of SERT protein levels in the DRN and three brain regions were attenuated by simultaneous treatment with fluoxetine, an antidepressant that specifically inhibits serotonin reuptake. The results indicate that up-regulation in SERT protein levels in the DRN and forebrain limbic structures caused by CSD regimen was mainly motivated by CORT through corticosteroid receptors. The present findings demonstrate that chronic stress is closely correlated with the serotonergic system by acting on the regulation of the SERT expression in the DRN and its projection regions, which may contribute to the development of depression. © 2012 International Society for Neurochemistry.

  15. 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.


    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.

  16. The conserved N-terminus of human rhinovirus capsid protein VP4 contains membrane pore-forming activity and is a target for neutralizing antibodies. (United States)

    Panjwani, Anusha; Asfor, Amin S; Tuthill, Tobias J


    Human rhinovirus is the causative agent of the common cold and belongs to the non-enveloped picornavirus family. A trigger such as receptor binding or low pH initiates conformational changes in the capsid that allow the virus to attach to membranes and form a pore for the translocation of viral RNA into the cytoplasm. We previously showed that recombinant capsid protein VP4 was able to form membrane pores. In this study, we show the N-terminus but not C-terminus of VP4 formed pores with properties similar to full-length VP4 and consistent with the size required for transfer of RNA. Sera against the N-terminus but not C-terminus of VP4 were shown to neutralize virus infectivity. Together, this suggests that the N-terminus of VP4 is responsible for membrane activity. This study contributes to an improved understanding of the mechanisms for involvement of VP4 in entry and its potential as an antiviral target.

  17. The role of the C-terminus of human α-synuclein: intra-disulfide bonds between the C-terminus and other regions stabilize non-fibrillar monomeric isomers. (United States)

    Hong, Dong-Pyo; Xiong, Wei; Chang, Jui-Yoa; Jiang, Chuantao


    Substantial evidence implicates that the aggregation of α-synuclein (αSyn) is a critical factor in the pathogenesis of Parkinson's disease. This study focuses on the role of αSyn C-terminus. We introduced two additional cysteine residues at positions 107 and 124 (A107C and A124C) to our previous construct. Five X-isomers of oxidative-folded mutation of α-synuclein with three disulfides were isolated and their secondary structures and aggregating features were analyzed. All isomers showed similar random coil structures as wild-type α-synuclein. However, these isomers did not form aggregates or fibrils, even with prolonged incubation, suggesting that the interactions between the C-terminal and N-terminal or central NAC region are important in maintaining the natively unfolded structure of αSyn and thus prevent αSyn from changing conformation, which is a critical step for fibrillation. Published by Elsevier B.V.

  18. Quantitative investigation of the affinity of human respiratory syncytial virus phosphoprotein C-terminus binding to nucleocapsid protein. (United States)

    Shapiro, Adam B; Gao, Ning; O'Connell, Nichole; Hu, Jun; Thresher, Jason; Gu, Rong-Fang; Overman, Ross; Hardern, Ian M; Sproat, Graham G


    There are no approved small molecule drug therapies for human respiratory syncytial virus (hRSV), a cause of morbidity and mortality in at-risk newborns, the immunocompromised, and the elderly. We have investigated as a potential novel hRSV drug target the protein-protein interaction between the C-terminus of the viral phosphoprotein (P) and the viral nucleocapsid protein (N), components of the ribonucleoprotein complex that contains, replicates, and transcribes the viral RNA genome. Earlier work by others established that the 9 C-terminal residues of P are necessary and sufficient for binding to N. We used a fluorescence anisotropy assay, surface plasmon resonance and 2-D NMR to quantify the affinities of peptides based on the C terminus of P for RNA-free, monomeric N-terminal-truncated N(13-391). We calculated the contributions to the free energies of binding of P to N(13-391) attributable to the C-terminal 11 residues, phosphorylation of the C-terminal 2 serine residues, the C-terminal Asp-Phe, and the phenyl ring of the C-terminal Phe. Binding studies confirmed the crucial role of the phosphorylated C-terminal peptide D(pS)DNDL(pS)LEDF for binding of P to RNA-free, monomeric N(13-391), contributing over 90% of the binding free energy at low ionic strength. The phenyl ring of the C-terminal Phe residue contributed an estimated -2.7 kcal/mole of the free energy of binding, the C-terminal Asp-Phe residues contributed -3.8 kcal/mole, the sequence DSDNDLSLE contributed -3.1 kcal/mole, and phosphorylation of the 2 Ser residues contributed -1.8 kcal/mole. Due to the high negative charge of the C-terminal peptide, the affinity of the P C-terminus for N(13-391) decreased as the ionic strength increased. The results support the idea that the interaction of the C-terminal residues of P with N constitutes a protein-protein interaction hotspot that may be a suitable target for small-molecule drugs that inhibit viral genome replication and transcription.

  19. The long N-terminus of the C. elegans DNA repair enzyme APN-1 targets the protein to the nucleus of a heterologous system. (United States)

    Wang, Zhiqiang; Yang, Xiaoming; Mazouzi, Abdelghani; Ramotar, Dindial


    We previously isolated from a Caenorhabditis elegans cDNA library, designed for two-hybrid screening, a gene encoding the DNA repair enzyme APN-1 using cross-specie complementation analysis of the Saccharomyces cerevisiae apn1∆ apn2∆ tpp1∆ triple mutant deficient in the ability to repair several types of DNA lesions including apurinic/apyrimidinic (AP) sites. We subsequently purified the APN-1 from this yeast mutant and demonstrated that it possesses four distinct DNA repair activities. However, following the re-annotation of the C. elegans genome we discovered that the functionally active APN-1 encoded by the cDNA from the library might lack 108 amino acid residues from the N-terminus. We therefore synthesized the entire C. elegans apn-1 gene encoding the putative full-length APN-1 and created several N-terminal deletion mutants lacking either 63, 83 or 118 amino acid residues. The full-length APN-1, APN-1 (1-63Δ) and APN-1 (1-83Δ), but not APN-1 (1-118Δ) were stably expressed in the yeast triple mutant and cleaved the AP site substrate. However, only the full-length APN-1 rescued the yeast mutant from the genotoxicity caused by methyl methane sulfonate, a DNA damaging agent that creates AP sites in the genome. The full-length APN-1 was localized to the yeast nucleus, while APN-1 (1-63Δ) and APN-1 (1-83Δ) retained a cytoplasmic distribution. Our data suggest that the N-terminal region has no direct role in the DNA repair functions of APN-1 other than to target the protein to the nucleus and possibly to maintain its stability. Thus, the truncated APN-1, previously isolated from the two-hybrid library, ability to complement the yeast triple mutant depends on the engineered SV40 nuclear localization signal. Copyright © 2014 Elsevier B.V. All rights reserved.


    Directory of Open Access Journals (Sweden)

    Oleksandr V. Tebenko


    Full Text Available Tools for sites building that offer users the ability to work together, an actual theme in information society and modern Web technologies. This article considers the SharePoint system, which enables to create sites of any complexity, including large portals with a complex structure of documents. Purpose of this article is to consider the main points of site creating and its setting with tools of SharePoint system, namely: a site template creating and configuring, web application environment to create and configure Web applications, change of existing and creation of new theme site, a web part setting.

  1. Creating a family health history (United States)

    Family health history; Create a family health history; Family medical history ... include your: Genes Diet and exercise habits Environment Family members tend to share certain behaviors, genetic traits, ...

  2. Importance of Residue 13 and the C-Terminus for the Structure and Activity of the Antimicrobial Peptide Aurein 2.2

    DEFF Research Database (Denmark)

    Cheng, John T.J.; Hale, John D.; Kindrachuk, Jason


    . In addition, we investigated a number of peptides with truncations at the C-terminus to determine whether the C-terminus, which contains residue 13, is crucial for antimicrobial activity. Solution circular dichroism results demonstrated that the L13F mutation and the truncation of the C-terminus by six...... of the peptide to insert into lipid bilayers. 31P NMR spectroscopy showed that all peptides disorder the headgroups. The implications of these results in terms of antimicrobial activity and the ability of these peptides to induce leakage in S. aureus are discussed. The results suggest that the presence of the 13......Previous studies on aurein 2.2 and 2.3 in DMPC/DMPG and POPC/POPG membranes have shown that bilayer thickness and phosphatidylglycerol content have a significant impact on the interaction of these peptides with membrane bilayers. Further examination with the DiSC35 assay has indicated that aurein 2...

  3. 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


    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...... that the cytoplasmic tail domain of US28 per se regulates receptor endocytosis, independent of the signaling ability of the core domain of US28. The constitutive endocytic property of the US28 c-tail was transposable to other 7TM receptors, the herpes virus 8-encoded ORF74 and the tachykinin NK1 receptor (ORF74-US28......-ctail and NK1-US28-ctail). Deletion of the US28 C terminus resulted in reduced constitutive endocytosis and consequently enhanced signaling capacity of all receptors tested as assessed by inositol phosphate turnover, NF-kappa B, and cAMP-responsive element-binding protein transcription assays. We...

  4. The C-terminus of kinesin-14 Ncd is a crucial component of the force generating mechanism. (United States)

    Szczęsna, Ewa; Kasprzak, Andrzej A


    Ncd, a member of kinesin-14 family motors, uses the power stroke, a lever-like pivoting action of a long and stiff element, to exert force and generate movement. To better understand the role of the Ncd C-terminus in this process we produced four Ncd mutants in which this segment was altered or deleted. For these proteins we measured their affinity to the microtubule, steady-state ATPase and gliding velocity in multiple motor assays. The mutations had a dramatic effect on all three parameters measured, suggesting that the C-terminal residues of Ncd play an important role in modulating the interaction of the motor with the microtubule. Copyright © 2012 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.

  5. Pan-ice-sheet glacier terminus change in East Antarctica reveals sensitivity of Wilkes Land to sea-ice changes. (United States)

    Miles, Bertie W J; Stokes, Chris R; Jamieson, Stewart S R


    The dynamics of ocean-terminating outlet glaciers are an important component of ice-sheet mass balance. Using satellite imagery for the past 40 years, we compile an approximately decadal record of outlet-glacier terminus position change around the entire East Antarctic Ice Sheet (EAIS) marine margin. We find that most outlet glaciers retreated during the period 1974-1990, before switching to advance in every drainage basin during the two most recent periods, 1990-2000 and 2000-2012. The only exception to this trend was in Wilkes Land, where the majority of glaciers (74%) retreated between 2000 and 2012. We hypothesize that this anomalous retreat is linked to a reduction in sea ice and associated impacts on ocean stratification, which increases the incursion of warm deep water toward glacier termini. Because Wilkes Land overlies a large marine basin, it raises the possibility of a future sea level contribution from this sector of East Antarctica.

  6. Pan–ice-sheet glacier terminus change in East Antarctica reveals sensitivity of Wilkes Land to sea-ice changes (United States)

    Miles, Bertie W. J.; Stokes, Chris R.; Jamieson, Stewart S. R.


    The dynamics of ocean-terminating outlet glaciers are an important component of ice-sheet mass balance. Using satellite imagery for the past 40 years, we compile an approximately decadal record of outlet-glacier terminus position change around the entire East Antarctic Ice Sheet (EAIS) marine margin. We find that most outlet glaciers retreated during the period 1974–1990, before switching to advance in every drainage basin during the two most recent periods, 1990–2000 and 2000–2012. The only exception to this trend was in Wilkes Land, where the majority of glaciers (74%) retreated between 2000 and 2012. We hypothesize that this anomalous retreat is linked to a reduction in sea ice and associated impacts on ocean stratification, which increases the incursion of warm deep water toward glacier termini. Because Wilkes Land overlies a large marine basin, it raises the possibility of a future sea level contribution from this sector of East Antarctica. PMID:27386519

  7. Creating

    DEFF Research Database (Denmark)

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


    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...

  8. Functional Consequences of Deletions of the N Terminus of the [epsilon] Subunit of the Chloroplast ATP Synthase. (United States)

    Cruz, J. A.; Radkowski, C. A.; McCarty, R. E.


    The [epsilon] subunit of the chloroplast ATP synthase functions in part to prevent wasteful ATP hydrolysis by the enzyme. In addition, [epsilon] together with the remainder of the catalytic portion of the synthase (CF1) is required to block the nonproductive leak of protons through the membrane-embedded component of the synthase (CFO). Mutant [epsilon] subunits of the spinach (Spinacia oleracea) chloroplast ATP synthase that lack 5, 11, or 20 amino acids from their N termini ([epsilon]-[delta]5N, [epsilon]-[delta]11N, and [epsilon]-[delta]20N, respectively), were overexpressed as inclusion bodies. Using a procedure that resulted in the folding of full-length, recombinant [epsilon] in a biologically active form, none of these truncated forms resulted in [epsilon] that inhibited the ATPase activity of CF1 deficient in [epsilon], CF1(-[epsilon]). Yet, the [epsilon]-[delta]5N and [epsilon]-[delta]11N peptides significantly inhibited the ATPase activity of CF1(-[epsilon]) bound to CFO in NaBr-treated thylakoids. Although full-length [epsilon] rapidly inhibited the ATPase activity of CF1(-[epsilon]) in solution or bound to CFO, an extended period was required for the truncated forms to inhibit membrane-bound CF1(-[epsilon]). Despite the fact that [epsilon]-[delta]5N significantly inhibited the ATPase activity of CF1(-[epsilon]) bound to CFO, it did not block the proton conductance through CFO in NaBr-treated thylakoids reconstituted with CF1(-[epsilon]). Based on selective proteolysis and the binding of 8-anilino-1-naphthalene sulfonic acid, each of the truncated peptides gained significant secondary structure after folding. These results strongly suggest (a) that the N terminus of [epsilon] is important in its binding to CF1, (b) that CF0 stabilizes [epsilon] binding to the entire ATP synthase, and (c) that the N terminus may play some role in the regulation of proton flux through CFO.

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

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


    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. Ferritin structure from Mycobacterium tuberculosis: comparative study with homologues identifies extended C-terminus involved in ferroxidase activity.

    Directory of Open Access Journals (Sweden)

    Garima Khare

    Full Text Available Ferritins are recognized as key players in the iron storage and detoxification processes. Iron acquisition in the case of pathogenic bacteria has long been established as an important virulence mechanism. Here, we report a 3.0 Å crystal structure of a ferritin, annotated as Bacterioferritin B (BfrB, from Mycobacterium tuberculosis (Mtb, the causative agent of tuberculosis that continues to be one of the world's deadliest diseases. Similar to the other members of ferritin family, the Mtb BfrB subunit exhibits the characteristic fold of a four-helical bundle that possesses the ferroxidase catalytic centre. We compare the structure of Mtb BfrB with representatives of the ferritin family belonging to the archaea, eubacteria and eukarya. Unlike most other ferritins, Mtb BfrB has an extended C-terminus. To dissect the role of this extended C-terminus, truncated Mtb BfrB was purified and biochemical studies implicate this region in ferroxidase activity and iron release in addition to providing stability to the protein. Functionally important regions in a protein of known 3D-structure can be determined by estimating the degree of conservation of the amino-acid sites with its close homologues. Based on the comparative studies, we identify the slowly evolving conserved sites as well as the rapidly evolving variable sites and analyze their role in relation to structure and function of Mtb BfrB. Further, electrostatic computations demonstrate that although the electrostatic environment of catalytic residues is preserved within the family, extensive variability is exhibited by residues defining the channels and pores, in all likelihood keeping up with the diverse functions executed by these ferritins in varied environments.

  11. The C Terminus of the Catalytic Domain of Type A Botulinum Neurotoxin May Facilitate Product Release from the Active Site* (United States)

    Mizanur, Rahman M.; Frasca, Verna; Swaminathan, Subramanyam; Bavari, Sina; Webb, Robert; Smith, Leonard A.; Ahmed, S. Ashraf


    Botulinum neurotoxins are the most toxic of all compounds. The toxicity is related to a poor zinc endopeptidase activity located in a 50-kDa domain known as light chain (Lc) of the toxin. The C-terminal tail of Lc is not visible in any of the currently available x-ray structures, and it has no known function but undergoes autocatalytic truncations during purification and storage. By synthesizing C-terminal peptides of various lengths, in this study, we have shown that these peptides competitively inhibit the normal catalytic activity of Lc of serotype A (LcA) and have defined the length of the mature LcA to consist of the first 444 residues. Two catalytically inactive mutants also inhibited LcA activity. Our results suggested that the C terminus of LcA might interact at or near its own active site. By using synthetic C-terminal peptides from LcB, LcC1, LcD, LcE, and LcF and their respective substrate peptides, we have shown that the inhibition of activity is specific only for LcA. Although a potent inhibitor with a Ki of 4.5 μm, the largest of our LcA C-terminal peptides stimulated LcA activity when added at near-stoichiometric concentration to three versions of LcA differing in their C-terminal lengths. The result suggested a product removal role of the LcA C terminus. This suggestion is supported by a weak but specific interaction determined by isothermal titration calorimetry between an LcA C-terminal peptide and N-terminal product from a peptide substrate of LcA. Our results also underscore the importance of using a mature LcA as an inhibitor screening target. PMID:23779108

  12. Creating Our Own Online Community

    Directory of Open Access Journals (Sweden)

    Mihaela TUTUNEA


    Full Text Available Creating our own online community is easy to do, by welcoming those who have an active presence online; first of all, we must have a well developed strategy of our own "empire", starting from the idea of creating the final benefit for our cyber-consumers.

  13. Creating Student-Friendly Tests (United States)

    Salend, Spencer J.


    Creating a fair, reliable, teacher-made test is a challenge. Every year poorly designed tests fail to accurately measure many students' learning--and negatively affect their academic futures. Salend, a well-known writer on assessment for at-risk students who consults with schools on assessment procedures, offers guidelines for creating tests that…

  14. 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


    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

  15. Industrial Engineering: creating a network!


    Prado-Prado, José Carlos


    [EN] This paper presents a brief history of the Industrial Engineering Conference (CIO), and specially reinforces the role of the CIOs as a forum for building a network and creating log-term relationships Prado-Prado, JC. (2016). Industrial Engineering: creating a network!. International Journal of Production Management and Engineering. 4(2):41-42. doi:10.4995/ijpme.2016.5964. 41 42 4 2

  16. Creating visual explanations improves learning. (United States)

    Bobek, Eliza; Tversky, Barbara


    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.

  17. Functional evolution of the photolyase/cryptochrome protein family: importance of the C terminus of mammalian CRY1 for circadian core oscillator performance.

    NARCIS (Netherlands)

    I. Chaves (Ines); K. Yagita (Kazuhiro); S. Barnhoorn (Sander); H. Okamura (Hitoshi); G.T.J. van der Horst (Gijsbertus); F. Tamanini (Filippo)


    textabstractCryptochromes (CRYs) are composed of a core domain with structural similarity to photolyase and a distinguishing C-terminal extension. While plant and fly CRYs act as circadian photoreceptors, using the C terminus for light signaling, mammalian CRY1 and CRY2 are integral components of

  18. Role of the cytosolic loop C2 and the C-terminus of YidC in ribosome binding and insertion activity

    NARCIS (Netherlands)

    Geng, Yanping; Kedrov, Alexej; Caumanns, Joseph J; Crevenna, Alvaro H; Lamb, Don C; Beckmann, Roland; Driessen, Arnold J M


    Members of the YidC/Oxa1/Alb3 protein family mediate membrane protein insertion and this process is initiated by the assembly of YidC:ribosome nascent chain (RNC) complexes at the inner leaflet of the lipid bilayer. The positively charged C-terminus of Escherichia coli YidC plays a significant role

  19. The Daughterless N-terminus directly mediates synergistic interactions with Notch transcription complexes via the SPS+A DNA transcription code

    Directory of Open Access Journals (Sweden)

    Xia Li


    Full Text Available Abstract Background Cell-specific expression of a subset of Enhancer of split (E(spl-C genes in proneural clusters is mediated by synergistic interactions between bHLH A (basic Helix-Loop-Helix Activator and Notch-signalling transcription complex (NTC proteins. For a some of these E(spl-C genes, such as m8, these synergistic interactions are programmed by an "SPS+A" transcription code in the cis-regulatory regions. However, the molecular mechanisms underlying this synergistic interaction between NTCs and proneural bHLH A proteins are not fully understood. Findings Using cell transcription assays, we show that the N-terminal region of the Daughterless (Da bHLH A protein is critical for synergistic interactions with NTCs that activate the E(spl-C m8 promoter. These assays also show that this interaction is dependent on the specific inverted repeat architecture of Suppressor of Hairless (Su(H binding sites in the SPS+A transcription code. Using protein-protein interaction assays, we show that two distinct regions within the Da N-terminus make a direct physical interaction with the NTC protein Su(H. Deletion of these interaction domains in Da creates a dominant negative protein that eliminates NTC-bHLH A transcriptional synergy on the m8 promoter. In addition, over-expression of this dominant negative Da protein disrupts Notch-mediated lateral inhibition during mechanosensory bristle neurogenesis in vivo. Conclusion These findings indicate that direct physical interactions between Da-N and Su(H are critical for the transcriptional synergy between NTC and bHLH A proteins on the m8 promoter. Our results also indicate that the orientation of the Su(H binding sites in the SPS+A transcription code are critical for programming the interaction between Da-N and Su(H proteins. Together, these findings provide insight into the molecular mechanisms by which the NTC synergistically interacts with bHLH A proteins to mediate Notch target gene expression in

  20. The Membrane M Protein Carboxy Terminus Binds to Transmissible Gastroenteritis Coronavirus Core and Contributes to Core Stability (United States)

    Escors, David; Ortego, Javier; Laude, Hubert; Enjuanes, Luis


    The architecture of transmissible gastroenteritis coronavirus includes three different structural levels, the envelope, an internal core, and the nucleocapsid that is released when the core is disrupted. Starting from purified virions, core structures have been reproducibly isolated as independent entities. The cores were stabilized at basic pH and by the presence of divalent cations, with Mg2+ ions more effectively contributing to core stability. Core structures showed high resistance to different concentrations of detergents, reducing agents, and urea and low concentrations of monovalent ions (<200 mM). Cores were composed of the nucleoprotein, RNA, and the C domain of the membrane (M) protein. At high salt concentrations (200 to 300 mM), the M protein was no longer associated with the nucleocapsid, which resulted in destruction of the core structure. A specific ionic interaction between the M protein carboxy terminus and the nucleocapsid was demonstrated using three complementary approaches: (i) a binding assay performed between a collection of M protein amino acid substitution or deletion mutants and purified nucleocapsids that led to the identification of a 16-amino-acid (aa) domain (aa 237 to 252) as being responsible for binding the M protein to the nucleocapsid; (ii) the specific inhibition of this binding by monoclonal antibodies (MAbs) binding to a carboxy-terminal M protein domain close to the indicated peptide but not by MAbs specific for the M protein amino terminus; and (iii) a 26-residue peptide, including the predicted sequence (aa 237 to 252), which specifically inhibited the binding. Direct binding of the M protein to the nucleoprotein was predicted, since degradation of the exposed RNA by RNase treatment did not affect the binding. It is proposed that the M protein is embedded within the virus membrane and that the C region, exposed to the interior face of the virion in a population of these molecules, interacts with the nucleocapsid to which it

  1. Effect of adding amino acids residues in N- and C-terminus of Vip3Aa16 (L121I) toxin. (United States)

    Sellami, Sameh; Cherif, Marwa; Jamoussi, Kaïs


    To study the importance of N- and C-terminus of Bacillus thuringiensis Vip3Aa16 (L121I) toxin (88 kDa), a number of mutants were generated. The addition of two (2R: RS) or eleven (11R: RSRPGHHHHHH) amino acid residues at the Vip3Aa16 (L121I) C-terminus allowed to an unappropriated folding illustrated by the abundant presence of the 62 kDa proteolytic form. The produced Vip3Aa16 (L121I) full length form was less detected when increasing the number of amino acids residues in the C-terminus. Bioassays demonstrated that the growth of the lepidopteran Ephestia kuehniella was slightly affected by Vip3Aa16 (L121I)-2R and not affected by Vip3Aa16 (L121I)-11R. Additionally, the fusion at the Vip3Aa16 (L121I) N-terminus of 39 amino acids harboring the E. coli OmpA leader peptide and the His-tag sequence allowed to the increase of protease sensitivity of Vip3Aa16 (L121I) full length form, as only the 62 kDa proteolysis form was detected. Remarkably, this fused protein produced in Escherichia coli (E. coli) was biologically inactive toward Ephestia kuehniella larvae. Thus, the N-terminus of the protein is required to the accomplishment of the insecticidal activity of Vip3 proteins. This report serves as guideline for the study of Vip3Aa16 (L121I) protein stability and activity. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  2. The DNA damage response and checkpoint adaptation in Saccharomyces cerevisiae: distinct roles for the replication protein A2 (Rfa2) N-terminus. (United States)

    Ghospurkar, Padmaja L; Wilson, Timothy M; Severson, Amber L; Klein, Sarah J; Khaku, Sakina K; Walther, André P; Haring, Stuart J


    In response to DNA damage, two general but fundamental processes occur in the cell: (1) a DNA lesion is recognized and repaired, and (2) concomitantly, the cell halts the cell cycle to provide a window of opportunity for repair to occur. An essential factor for a proper DNA-damage response is the heterotrimeric protein complex Replication Protein A (RPA). Of particular interest is hyperphosphorylation of the 32-kDa subunit, called RPA2, on its serine/threonine-rich amino (N) terminus following DNA damage in human cells. The unstructured N-terminus is often referred to as the phosphorylation domain and is conserved among eukaryotic RPA2 subunits, including Rfa2 in Saccharomyces cerevisiae. An aspartic acid/alanine-scanning and genetic interaction approach was utilized to delineate the importance of this domain in budding yeast. It was determined that the Rfa2 N-terminus is important for a proper DNA-damage response in yeast, although its phosphorylation is not required. Subregions of the Rfa2 N-terminus important for the DNA-damage response were also identified. Finally, an Rfa2 N-terminal hyperphosphorylation-mimetic mutant behaves similarly to another Rfa1 mutant (rfa1-t11) with respect to genetic interactions, DNA-damage sensitivity, and checkpoint adaptation. Our data indicate that post-translational modification of the Rfa2 N-terminus is not required for cells to deal with "repairable" DNA damage; however, post-translational modification of this domain might influence whether cells proceed into M-phase in the continued presence of unrepaired DNA lesions as a "last-resort" mechanism for cell survival. Copyright © 2015 by the Genetics Society of America.

  3. Purification method for recombinant proteins based on a fusion between the target protein and the C-terminus of calmodulin (United States)

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


    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.

  4. Cloning and characterization of carboxyl terminus of heat shock cognate 70-interacting protein gene from the silkworm, Bombyx mori. (United States)

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


    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.

  5. 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.; Burkhart, William A.; Consler, Thomas G.; Cowan, David J.; Gottschalk, William K.; Miller, Aaron B; Short, Steven A.; Tran, Thuy B.; Williams, Shawn P.; (GSKNC); (Duke); (UNC)


    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 -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.

  6. Intra-arterial therapy for cardio embolic internal carotid artery terminus occlusion: The past and present status in real practice

    Energy Technology Data Exchange (ETDEWEB)

    Jung, Seung Guk; Jung, Cheol Kyu; KIm, Jae Hyoung; Choi, Byung Se; Kim, Beomn Joon; Han, Moon Ku; Bae, Hee Joon [Seoul National University Bundang Hospital, Seongnam (Korea, Republic of); Kwon, Bae Ju [Dept. of Radiology, Myongji Hospital, Goyang (Korea, Republic of); Cha, Sang Hoon [Dept. of Radiology, Chungbuk National University College of Medicine, Daejeon (Korea, Republic of)


    There is little data on the effect of intra-arterial therapy (IAT) in acute cardioembolic internal carotid artery terminus (ICAT) occlusion that has poor prognosis. We determined procedural and clinical outcomes in patients with acute cardioembolic ICAT occlusion treated with different methods of IAT. On retrospective review of our registry, patients with cardioembolic ICAT occlusion were categorized as thrombolytic-based IAT group (TLG) and thrombectomy-based IAT group (TEG) according to the primary endovascular technique. Subsequently, procedural and clinical outcomes were compared. Fifty-five patients had cardioembolic ICAT occlusion and 18 patients were assigned to TLG and 37 patients to TEG. The rate of complete reperfusion was significantly higher and the groin puncture to reperfusion time was significantly shorter in TEG than those in TLG. There was a trend towards functional outcome at 3 months in the TEG group; however, it was not statistically significant (p = 0.06). Age, baseline Albert Stroke program early CT score and puncture to reperfusion time were factors affecting unfavorable outcome at 3 months, on multivariable analysis. Thrombectomy-based IAT has advantages over thrombolytic-based IAT in terms of the reduction of groin puncture to reperfusion time and improvement of the rate of complete reperfusion.

  7. 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.

  8. 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: [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: [Pediatric Endocrine Unit, MassGeneral Hospital for Children and Harvard Medical School, Boston, Massachusetts, 02114-2696 (United States)


    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.

  9. Changes in the accessibility of the HIV-1 Integrase C-terminus in the presence of cellular proteins

    Directory of Open Access Journals (Sweden)

    Zanta-Boussif Maria-Antonietta


    Full Text Available Abstract Background Following entry, uncoating, and reverse transcription, a number of cellular proteins become associated with the Human Immunodeficiency Virus type 1 (HIV-1 pre-integration complex (PIC. With the goal of obtaining reagents for the analysis of the HIV-1 PIC composition and localisation, we have constructed functional integrase (IN and matrix (MA proteins that can be biotinylated during virus production and captured using streptavidin-coated beads. Results Although the labelled C-terminus allows for the sensitive detection of virion-associated IN, it becomes inaccessible in the presence of cellular proteins. This masking is not dependent on the nature of the tag and does not occur with the tagged MA. It was not observed either with an IN mutant unable to interact with LEDGF/p75, or when LEDGF/p75 was depleted from cells. Conclusion Our observation suggests that a structural rearrangement or oligomerization of the IN protein occurs during the early steps of infection and that this process is related to the presence of LEDGF/p75.

  10. Anchoring Intrinsically Disordered Proteins to Multiple Targets: Lessons from N-Terminus of the p53 Protein

    Directory of Open Access Journals (Sweden)

    Yongqi Huang


    Full Text Available Anchor residues, which are deeply buried upon binding, play an important role in protein–protein interactions by providing recognition specificity and facilitating the binding kinetics. Up to now, studies on anchor residues have been focused mainly on ordered proteins. In this study, we investigated anchor residues in intrinsically disordered proteins (IDPs which are flexible in the free state. We identified the anchor residues of the N-terminus of the p53 protein (Glu17–Asn29, abbreviated as p53N which are involved in binding with two different targets (MDM2 and Taz2, and analyzed their side chain conformations in the unbound states. The anchor residues in the unbound p53N were found to frequently sample conformations similar to those observed in the bound complexes (i.e., Phe19, Trp23, and Leu26 in the p53N-MDM2 complex, and Leu22 in the p53N-Taz2 complex. We argue that the bound-like conformations of the anchor residues in the unbound state are important for controlling the specific interactions between IDPs and their targets. Further, we propose a mechanism to account for the binding promiscuity of IDPs in terms of anchor residues and molecular recognition features (MoRFs.

  11. An androgen receptor NH2-terminal conserved motif interacts with the COOH terminus of the Hsp70-interacting protein (CHIP). (United States)

    He, Bin; Bai, Suxia; Hnat, Andrew T; Kalman, Rebecca I; Minges, John T; Patterson, Cam; Wilson, Elizabeth M


    The NH2-terminal sequence of steroid receptors is highly variable between different receptors and in the same receptor from different species. In this study, a primary sequence homology comparison identified a 14-amino acid NH2-terminal motif of the human androgen receptor (AR) that is common to AR from all species reported, including the lower vertebrates. The evolutionarily conserved motif is unique to AR, with the exception of a partial sequence in the glucocorticoid receptor of higher species. The presence of the conserved motif in AR and the glucocorticoid receptor and its absence in other steroid receptors suggests convergent evolution. The function of the AR NH2-terminal conserved motif was suggested from a yeast two-hybrid screen that identified the COOH terminus of the Hsp70-interacting protein (CHIP) as a binding partner. We found that CHIP functions as a negative regulator of AR transcriptional activity by promoting AR degradation. In support of this, two mutations in the AR NH2-terminal conserved motif previously identified in the transgenic adenocarcinoma of mouse prostate model reduced the interaction between CHIP and AR. Our results suggest that the AR NH2-terminal domain contains an evolutionarily conserved motif that functions to limit AR transcriptional activity. Moreover, we demonstrate that the combination of comparative sequence alignment and yeast two-hybrid screening using short conserved peptides as bait provides an effective strategy to probe the structure-function relationships of steroid receptor NH2-terminal domains and other intrinsically unstructured transcriptional regulatory proteins.

  12. Creating an Innovative Learning Organization (United States)

    Salisbury, Mark


    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…

  13. On Creating and Sustaining Alternatives

    DEFF Research Database (Denmark)

    Kyng, Morten


    (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...

  14. SPECIAL REPORT: Creating Conference Video

    Directory of Open Access Journals (Sweden)

    Noel F. Peden


    Full Text Available Capturing video at a conference is easy. Doing it so the product is useful is another matter. Many subtle problems come into play so that video and audio obtained can be used to create a final product. This article discusses what the author learned in the two years of shooting and editing video for Code4Lib conference.

  15. Creating Space for Children's Literature (United States)

    Serafini, Frank


    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…

  16. We create our own reality

    CERN Multimedia


    " 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).

  17. Creating legitimacy across international contexts

    DEFF Research Database (Denmark)

    Andersen, Poul Houman; Rask, Morten


    in Denmark, Israel, Canada, and Australia using expert interviews as well as content analysis of newspaper articles and other secondary sources. Storytelling, which is found to be central to the legitimacy-creating efforts of international business ventures, interacts with existing discourses in the diverse...

  18. Creating Presentations on ICT Classes (United States)

    Marchis, Iuliana


    The article focuses on the creation of presentations on ICT classes. The first part highlights the most important steps when creating a presentation. The main idea is, that the computer presentation shouldn't consist only from the technological part, i.e. the editing of the presentation in a computer program. There are many steps before and after…

  19. Creating speech-synchronized animation. (United States)

    King, Scott A; Parent, Richard E


    We present a facial model designed primarily to support animated speech. Our facial model takes facial geometry as input and transforms it into a parametric deformable model. The facial model uses a muscle-based parameterization, allowing for easier integration between speech synchrony and facial expressions. Our facial model has a highly deformable lip model that is grafted onto the input facial geometry to provide the necessary geometric complexity needed for creating lip shapes and high-quality renderings. Our facial model also includes a highly deformable tongue model that can represent the shapes the tongue undergoes during speech. We add teeth, gums, and upper palate geometry to complete the inner mouth. To decrease the processing time, we hierarchically deform the facial surface. We also present a method to animate the facial model over time to create animated speech using a model of coarticulation that blends visemes together using dominance functions. We treat visemes as a dynamic shaping of the vocal tract by describing visemes as curves instead of keyframes. We show the utility of the techniques described in this paper by implementing them in a text-to-audiovisual-speech system that creates animation of speech from unrestricted text. The facial and coarticulation models must first be interactively initialized. The system then automatically creates accurate real-time animated speech from the input text. It is capable of cheaply producing tremendous amounts of animated speech with very low resource requirements.

  20. A basic cluster in the N terminus of yellow fever virus NS2A contributes to infectious particle production. (United States)

    Voßmann, Stephanie; Wieseler, Janett; Kerber, Romy; Kümmerer, Beate Mareike


    The flavivirus NS2A protein is involved in the assembly of infectious particles. To further understand its role in this process, a charged-to-alanine scanning analysis was performed on NS2A encoded by an infectious cDNA clone of yellow fever virus (YFV). Fifteen mutants containing single, double, or triple charged-to-alanine changes were tested. Five of them did not produce infectious particles, whereas efficient RNA replication was detectable for two of the five NS2A mutants (R22A-K23A-R24A and R99A-E100A-R101A mutants). Prolonged cultivation of transfected cells resulted in the recovery of pseudorevertants. Besides suppressor mutants in NS2A, a compensating second-site mutation in NS3 (D343G) arose for the NS2A R22A-K23A-R24A mutant. We found this NS3 mutation previously to be suppressive for the NS2Aα cleavage site Q189S mutant, also deficient in virion assembly. In this study, the subsequently suggested interaction between NS2A and NS3 was proven by coimmunoprecipitation analyses. Using selectively permeabilized cells, we could demonstrate that the regions encompassing R22A-K23A-R24A and Q189S in NS2A are localized to the cytoplasm, where NS3 is also known to reside. However, the defect in particle production observed for the NS2A R22A-K23A-R24A and Q189S mutants was not due to a defect in physical interaction between NS2A and NS3, as the NS2A mutations did not interrupt NS3 interaction. In fact, a region just upstream of R22-K23-R24 was mapped to be critical for NS2A-NS3 interaction. Taken together, these data support a complex interplay between YFV NS2A and NS3 in virion assembly and identify a basic cluster in the NS2A N terminus to be critical in this process. Despite an available vaccine, yellow fever remains endemic in tropical areas of South America and Africa. To control the disease, antiviral drugs are required, and an understanding of the determinants of virion assembly is central to their development. In this study, we identified a basic cluster of

  1. Creating a climate for excellence. (United States)

    Lancaster, J


    Some people are motivated to achieve in a manner consistent with the goals of their organization while others pursue individual goals. The attitudes people hold determine their behavior. Therefore, the manager is charged with creating an environment that fosters employee commitment to organizational goals. To create a climate for achievement, managers must recognize that all employees want recognition. Employees perform more effectively when they understand the goals of the organization, know what is expected of them, and are part of a system that includes feedback and reinforcement. Generally, people perform more effectively in an environment with minimal threat and punishment; individual responsibility should be encouraged, rewards based on results, and a climate of trust and open communication should prevail.

  2. Creating advanced health informatics certification. (United States)

    Gadd, Cynthia S; Williamson, Jeffrey J; Steen, Elaine B; Fridsma, Douglas B


    In 2005, AMIA leaders and members concluded that certification of advanced health informatics professionals would offer value to individual practitioners, organizations that hire them, and society at large. AMIA's work to create advanced informatics certification began by leading a successful effort to create the clinical informatics subspecialty for American Board of Medical Specialties board-certified physicians. Since 2012, AMIA has been working to establish advanced health informatics certification (AHIC) for all health informatics practitioners regardless of their primary discipline. In November 2015, AMIA completed the first of 3 key tasks required to establish AHIC, with the AMIA Board of Directors' endorsement of proposed eligibility requirements. This AMIA Board white paper describes efforts to establish AHIC, reports on the current status of AHIC components, and provides a context for the proposed AHIC eligibility requirements. © The Author 2016. Published by Oxford University Press on behalf of the American Medical Informatics Association. All rights reserved. For Permissions, please email:

  3. Leadership Networking Connect, Collaborate, Create

    CERN Document Server

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


    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.

  4. Creating a Mobile Library Website (United States)

    Cutshall, Tom C.; Blake, Lindsay; Bandy, Sandra L.


    The overwhelming results were iPhones and Android devices. Since the library wasn't equipped technologically to develop an in-house application platform and because we wanted the content to work across all mobile platforms, we decided to focus on creating a mobile web-based platform. From the NLM page of mobile sites we chose the basic PubMed/…

  5. Creating and Sustaining Competitive Advantage


    Lage Hansen, Jakob


    The paper provides a general framework for examining how governance choice affects competitive advantage. I argue that firms rely on assets for competing, and that these assets can be accessed by different governance structures (i.e., they can be in- or outsourced). The transaction cost economics framework is used to expose strengths and weaknesses of governance structures with respect to creating and sustaining competitive advantage. The result is a tradeoff to consider when choosing how to ...

  6. The N-terminus of murine leukaemia virus p12 protein is required for mature core stability. (United States)

    Wight, Darren J; Boucherit, Virginie C; Wanaguru, Madushi; Elis, Efrat; Hirst, Elizabeth M A; Li, Wilson; Ehrlich, Marcelo; Bacharach, Eran; Bishop, Kate N


    The murine leukaemia virus (MLV) gag gene encodes a small protein called p12 that is essential for the early steps of viral replication. The N- and C-terminal regions of p12 are sequentially acting domains, both required for p12 function. Defects in the C-terminal domain can be overcome by introducing a chromatin binding motif into the protein. However, the function of the N-terminal domain remains unknown. Here, we undertook a detailed analysis of the effects of p12 mutation on incoming viral cores. We found that both reverse transcription complexes and isolated mature cores from N-terminal p12 mutants have altered capsid complexes compared to wild type virions. Electron microscopy revealed that mature N-terminal p12 mutant cores have different morphologies, although immature cores appear normal. Moreover, in immunofluorescent studies, both p12 and capsid proteins were lost rapidly from N-terminal p12 mutant viral cores after entry into target cells. Importantly, we determined that p12 binds directly to the MLV capsid lattice. However, we could not detect binding of an N-terminally altered p12 to capsid. Altogether, our data imply that p12 stabilises the mature MLV core, preventing premature loss of capsid, and that this is mediated by direct binding of p12 to the capsid shell. In this manner, p12 is also retained in the pre-integration complex where it facilitates tethering to mitotic chromosomes. These data also explain our previous observations that modifications to the N-terminus of p12 alter the ability of particles to abrogate restriction by TRIM5alpha and Fv1, factors that recognise viral capsid lattices.

  7. The N-terminus of murine leukaemia virus p12 protein is required for mature core stability.

    Directory of Open Access Journals (Sweden)

    Darren J Wight


    Full Text Available The murine leukaemia virus (MLV gag gene encodes a small protein called p12 that is essential for the early steps of viral replication. The N- and C-terminal regions of p12 are sequentially acting domains, both required for p12 function. Defects in the C-terminal domain can be overcome by introducing a chromatin binding motif into the protein. However, the function of the N-terminal domain remains unknown. Here, we undertook a detailed analysis of the effects of p12 mutation on incoming viral cores. We found that both reverse transcription complexes and isolated mature cores from N-terminal p12 mutants have altered capsid complexes compared to wild type virions. Electron microscopy revealed that mature N-terminal p12 mutant cores have different morphologies, although immature cores appear normal. Moreover, in immunofluorescent studies, both p12 and capsid proteins were lost rapidly from N-terminal p12 mutant viral cores after entry into target cells. Importantly, we determined that p12 binds directly to the MLV capsid lattice. However, we could not detect binding of an N-terminally altered p12 to capsid. Altogether, our data imply that p12 stabilises the mature MLV core, preventing premature loss of capsid, and that this is mediated by direct binding of p12 to the capsid shell. In this manner, p12 is also retained in the pre-integration complex where it facilitates tethering to mitotic chromosomes. These data also explain our previous observations that modifications to the N-terminus of p12 alter the ability of particles to abrogate restriction by TRIM5alpha and Fv1, factors that recognise viral capsid lattices.

  8. 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.

  9. 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.

  10. Distinct overlapping sequences at the carboxy-terminus of merlin regulate its tumour suppressor and morphogenic activity. (United States)

    Laulajainen, Minja; Melikova, Maria; Muranen, Taru; Carpén, Olli; Grönholm, Mikaela


    The Neurofibromatosis 2 (NF2) gene product merlin is a tumour suppressor, which in addition to inhibiting cell proliferation regulates cell morphology. The morphogenic properties of merlin may play a role in tumour suppression, as patient-derived tumour cells demonstrate cytoskeletal abnormalities. However, it is still unclear how these functions are linked. The N-terminal FERM-domain of merlin is highly homologous to the oncogenic protein ezrin, while the C-termini are less conserved, suggesting that the opposite effect of the proteins on proliferation could be mediated by their distinct C-terminal regions. In this study we characterize the role of the most C-terminal residues of merlin in the regulation of proliferation, cytoskeletal organization, phosphorylation and intramolecular associations. In addition to the two full-length merlin isoforms and truncating mutations found in patients, we focused on the evolutionally conserved C-terminal residues 545-547, also harbouring disease-causing mutations. We demonstrate that merlin induces cell extensions, which result from impaired retraction of protrusions rather than from increased formation of filopodia. The residues 538-568 were found particularly important for this morphogenic activity. The results further show that both merlin isoforms are able to equally inhibit proliferation, whereas C-terminal mutants affecting residues 545-547 are less effective in growth suppression. This study demonstrates that the C-terminus contains distinct but overlapping functional domains important for regulation of the morphogenic activity, intramolecular associations and cell proliferation. © 2012 The Authors Journal of Cellular and Molecular Medicine © 2012 Foundation for Cellular and Molecular Medicine/Blackwell Publishing Ltd.

  11. Deletion of the N-terminus of SF2/ASF permits RS-domain-independent pre-mRNA splicing. (United States)

    Shaw, Stephanie D; Chakrabarti, Sutapa; Ghosh, Gourisankar; Krainer, Adrian R


    Serine/arginine-rich (SR) proteins are essential splicing factors with one or two RNA-recognition motifs (RRMs) and a C-terminal arginine- and serine-rich (RS) domain. SR proteins bind to exonic splicing enhancers via their RRM(s), and from this position are thought to promote splicing by antagonizing splicing silencers, recruiting other components of the splicing machinery through RS-RS domain interactions, and/or promoting RNA base-pairing through their RS domains. An RS domain tethered at an exonic splicing enhancer can function as a splicing activator, and RS domains play prominent roles in current models of SR protein functions. However, we previously reported that the RS domain of the SR protein SF2/ASF is dispensable for in vitro splicing of some pre-mRNAs. We have now extended these findings via the identification of a short inhibitory domain at the SF2/ASF N-terminus; deletion of this segment permits splicing in the absence of this SR protein's RS domain of an IgM pre-mRNA substrate previously classified as RS-domain-dependent. Deletion of the N-terminal inhibitory domain increases the splicing activity of SF2/ASF lacking its RS domain, and enhances its ability to bind pre-mRNA. Splicing of the IgM pre-mRNA in S100 complementation with SF2/ASF lacking its RS domain still requires an exonic splicing enhancer, suggesting that an SR protein RS domain is not always required for ESE-dependent splicing activation. Our data provide additional evidence that the SF2/ASF RS domain is not strictly required for constitutive splicing in vitro, contrary to prevailing models for how the domains of SR proteins function to promote splicing.

  12. Biomedical scientist training officers' evaluation of integrated (co-terminus) Applied Biomedical Science BSc programmes: a multicentre study. (United States)

    Pitt, S J; Cunningham, J M


    The introduction of the Institute of Biomedical Science (IBMS) portfolio for pre-registration training in 2003 allowed universities to develop integrated (co-terminus) biomedical science BSc programmes. Students undertake structured placements within clinical pathology laboratories as part of their degree. The clinical training and professional development of students is undertaken by training officers (TOs), who are experienced Health Professions Council (HPC)-registered biomedical scientists and usually also members of the IBMS. This study aims to evaluate TOs' perceptions of these integrated degrees as a means of delivering pre-registration training for biomedical scientists. A questionnaire to collect quantitative data and be completed anonymously was sent to TOs, via staff at participating universities. Items considered TOs' perceptions in four categories: how well students fitted into the laboratory team, their professional and scientific development, the impact of delivering integrated degrees on service delivery, and the commitment to training students. Surveys took place in 2007, 2008 and 2009 and involved TOs taking students from 10, 14 and 17 universities each year, respectively. The response rates to the survey were 60% in 2007, 34% in 2008 and 12% in 2009. Participants were representative in terms of age, gender and pathology discipline and had a broad range of experience with students. The overall mean score for TOs perceptions was 3.38 in 2007 which increased significantly to 3.99 in 2009 (Kruskall Wallis test chi2 = 21.13, P<0.01). Mean scores in three of the four categories were positive in 2007, although the impact on service delivery was perceived negatively. In all areas, means were significantly greater in 2009. The results indicate that TOs view the integrated degrees favourably and are happy with the scientific and professional development of students. Although designing training sessions suitable for undergraduates took extra work initially

  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


    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. Targeting of the Dopamine Transporter Involves Discrete Epitopes in the Distal C Terminus But Does Not Require Canonical PDZ Domain Interactions

    DEFF Research Database (Denmark)

    Bjerggaard(Vægter), Christian; Fog, Jacob Ulrik; Hastrup, Hanne


    -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...... 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......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...

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

    DEFF Research Database (Denmark)

    Bjerggaard, Christian; Fog, Jacob U; Hastrup, Hanne


    -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...... 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......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...

  16. Interaction of a peptide derived from C-terminus of human TRPA1 channel with model membranes mimicking the inner leaflet of the plasma membrane. (United States)

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


    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.

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

    Energy Technology Data Exchange (ETDEWEB)

    Ambrose, R.L.; Mackenzie, J.M., E-mail:


    The West Nile virus strain Kunjin virus (WNV{sub KUN}) NS4A protein is a multifunctional protein involved in many aspects of the virus life-cycle and is a major component of the WNV{sub 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{sub 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.

  18. Seasonal and interannual variabilities in terminus position, glacier velocity, and surface elevation at Helheim and Kangerlussuaq Glaciers from 2008 to 2016 (United States)

    Kehrl, L. M.; Joughin, I.; Shean, D. E.; Floricioiu, D.; Krieger, L.


    The dynamic response of Greenland tidewater glaciers to oceanic and atmospheric change has varied both spatially and temporally. While some of this variability is likely related to regional climate signals, glacier geometry also appears to be important. In this study, we investigated the environmental and geometric controls on the seasonal and interannual evolution of Helheim and Kangerlussuaq Glaciers, Southeast Greenland, from 2008 to 2016, by combining year-round, satellite measurements of terminus position, glacier velocity, and surface elevation. While Helheim remained relatively stable with a lightly grounded terminus over this time period, Kangerlussuaq continued to lose mass as its grounding line retreated into deeper water. By summer 2011, Kangerlussuaq's grounding line had retreated into shallower water, and the glacier had an 5 km long floating ice tongue. We also observed seasonal variations in surface velocity and elevation at both glaciers. At Helheim, seasonal speedups and dynamic thinning occurred in the late summer when the terminus was most retreated. At Kangerlussuaq, we observed summer speedups due to surface-melt-induced basal lubrication and winter speedups due to ice-shelf retreat. We suggest that Helheim and Kangerlussuaq behaved differently on a seasonal timescale due to differences in the spatial extent of floating ice near their termini, which affected iceberg-calving behavior. Given that seasonal speedups and dynamic thinning can alter this spatial extent, these variations may be important for understanding the long-term evolution of these and other Greenland tidewater glaciers.

  19. Preventive Indicators for Creating Brownfields

    Directory of Open Access Journals (Sweden)

    Marija Burinskienė


    Full Text Available Although the problem of brownfields in urban territories is successfully limited, it is a negative phenomenon of a sustainable city. Moreover, the number of recently created brownfield territories has become higher than that of the regenerated ones. Such territories reduce the quality of the social and economic setting of a city as well as visually and physically affect the life quality of city residents. Unfortunately, methods for the revitalization of brownfield land have been applied to deal with the consequences of the problem rather than to limit the problem itself. The authors of the article have investigated the aspects to be avoided to not create brownfields. The indicators that enable predicting the probability of a territory becoming a brownfield have been analyzed in this paper. Countries develop and exist under different social and economic conditions. Therefore, there is no uniform and universally accepted system of indicators for brownfield prevention that can be applied in any country or city. The authors have attempted to implement a recently developed idea of indicators for prevention under Lithuanian conditions and have selected those facilitating the identification of brownfields with an aim of identifying the most significant ones warning about the potential harm from the creation of brownfields in Lithuania. The selected indicators have been grouped, taking into account social, economic, natural, building and infrastructure settings of the city and ranked by a group of experts in urban planning. The established hierarchy of indicators in the groups of urban setting has allowed the authors to select the most significant preventive indicators for brownfields. The created system of indicators could be applied in practice as a basis for monitoring pertinent data and tracking their change.

  20. Creating a digital medical illustration. (United States)

    Culley, Joanna


    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.

  1. Creating your own leadership brand. (United States)

    Kerfoot, Karlene


    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.


    Directory of Open Access Journals (Sweden)

    Umral Deveci


    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.

  3. 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


    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

  4. 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.

  5. Can the Universe create itself? (United States)

    Gott, J. Richard, III; Li, Li-Xin


    The question of first-cause has troubled philosophers and cosmologists alike. Now that it is apparent that our universe began in a big bang explosion, the question of what happened before the big bang arises. Inflation seems like a very promising answer, but as Borde and Vilenkin have shown, the inflationary state preceding the big bang could not have been infinite in duration-it must have had a beginning also. Where did it come from? Ultimately, the difficult question seems to be how to make something out of nothing. This paper explores the idea that this is the wrong question-that that is not how the Universe got here. Instead, we explore the idea of whether there is anything in the laws of physics that would prevent the Universe from creating itself. Because spacetimes can be curved and multiply connected, general relativity allows for the possibility of closed timelike curves (CTCs). Thus, tracing backwards in time through the original inflationary state we may eventually encounter a region of CTCs-giving no first-cause. This region of CTCs may well be over by now (being bounded toward the future by a Cauchy horizon). We illustrate that such models-with CTCs-are not necessarily inconsistent by demonstrating self-consistent vacuums for Misner space and a multiply connected de Sitter space in which the renormalized energy-momentum tensor does not diverge as one approaches the Cauchy horizon and solves Einstein's equations. Some specific scenarios (out of many possible ones) for this type of model are described. For example, a metastable vacuum inflates producing an infinite number of (big-bang-type) bubble universes. In many of these, either by natural causes or by action of advanced civilizations, a number of bubbles of metastable vacuum are created at late times by high energy events. These bubbles will usually collapse and form black holes, but occasionally one will tunnel to create an expanding metastable vacuum (a baby universe) on the other side of the

  6. Creating Interdisciplinarity within Monodisciplinary Structures

    DEFF Research Database (Denmark)

    Lindvig, Katrine

    The objectives of the PhD project were to explore the linkages between interdisciplinary research and education, and to follow the concrete development and execution of interdisciplinary educational activities. In order to meet these objectives, an extensive literature study and a two......-year ethnographic fieldwork were conducted. The PhD project was part of the development project ‘Interdisciplinary education at UCPH’, with the aim of improving and supporting interdisciplinary teaching and learning at the University of Copenhagen (UCPH). The findings of the PhD study point towards wide...... 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. Creating Genetic Resistance to HIV (United States)

    Burnett, John C.; Zaia, John A.; Rossi, John J.


    HIV/AIDS remains a chronic and incurable disease, in spite of the notable successes of highly active antiretroviral therapy. Gene therapy offers the prospect of creating genetic resistance to HIV that supplants the need for antiviral drugs. In sight of this goal, a variety of anti-HIV genes have reached clinical testing, including gene-editing enzymes, protein-based inhibitors, and RNA-based therapeutics. Combinations of therapeutic genes against viral and host targets are designed to improve the overall antiviral potency and reduce the likelihood of viral resistance. In cell-based therapies, therapeutic genes are expressed in gene modified T lymphocytes or in hematopoietic stem cells that generate an HIV-resistant immune system. Such strategies must promote the selective proliferation of the transplanted cells and the prolonged expression of therapeutic genes. This review focuses on the current advances and limitations in genetic therapies against HIV, including the status of several recent and ongoing clinical studies. PMID:22985479

  8. Creating and Recreating. Microcomputing Working Paper Series. (United States)

    Arms, Valarie

    This document describes a software program, CREATE, which was developed at Drexel University to guide students in creating English compositions. A second program, ReCREATE, guides students in reading their finished papers and making global revisions. CREATE asks 20 questions that a teacher might ask a student in a pre-writing conference. Unlike…

  9. Creating responsible partnerships in tourism

    Directory of Open Access Journals (Sweden)

    Ana Spitzer


    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. Adolescents and HIV: creating partnerships. (United States)

    Tierney, S


    Despite the President's directive on youth and HIV in 1997 to focus the nation's attention on adolescents and the battle against AIDS, prevention programs continue to be ineffective. The number of seropositive youth, ages 13 to 24 years old, is unclear due to inconsistent definitions of age ranges and inadequate access to testing. Youth have not sought testing for many reasons, including failing to perceive their vulnerability to HIV, confidentiality concerns, and not realizing the effectiveness of early treatment. Adolescents are creating independence, establishing relationships, and learning about drugs and alcohol. Young gay and bisexual men, drug-using youth, and youth of color are at high risk of HIV transmission. Identifying the population involved in risk-taking behavior and eliminating the behavior is an ineffective strategy for adolescent HIV prevention programs. Complicating the issue further, the goals and expectations of adolescents differ from the adults who design and deliver prevention programs. HIV education and prevention efforts need to address solutions to hopelessness, isolation, and violence, rather than focusing on the negative effects risky behaviors will have in the future. Effective programs combine a youth/adult partnership to take advantage of the strengths of each individual. Strategies for implementing prevention programs that address the specific needs of adolescents are suggested.

  11. Creating healthy and just bioregions. (United States)

    Pezzoli, Keith; Leiter, Robert Allen


    Dramatic changes taking place locally, regionally, globally, demand that we rethink strategies to improve public health, especially in disadvantaged communities where the cumulative impacts of toxicant exposure and other environmental and social stressors are most damaging. The emergent field of Sustainability Science, including a new bioregionalism for the 21st Century, is giving rise to promising place-based (territorially rooted) approaches. Embedded in this bioregional approach is an integrated planning framework (IPF) that enables people to map and develop plans and strategies that cut across various scales (e.g. from regional to citywide to neighborhood scale) and various topical areas (e.g. urban land use planning, water resource planning, food systems planning and "green infrastructure" planning) with the specific intent of reducing the impacts of toxicants to public health and the natural environment. This paper describes a case of bioregionally inspired integrated planning in San Diego, California (USA). The paper highlights food-water-energy linkages and the importance of "rooted" community-university partnerships and knowledge-action collaboratives in creating healthy and just bioregions.

  12. Laser Created Relativistic Positron Jets

    Energy Technology Data Exchange (ETDEWEB)

    Chen, H; Wilks, S C; Meyerhofer, D D; Bonlie, J; Chen, C D; Chen, S N; Courtois, C; Elberson, L; Gregori, G; Kruer, W; Landoas, O; Mithen, J; Murphy, C; Nilson, P; Price, D; Scheider, M; Shepherd, R; Stoeckl, C; Tabak, M; Tommasini, R; Beiersdorder, P


    Electron-positron jets with MeV temperature are thought to be present in a wide variety of astrophysical phenomena such as active galaxies, quasars, gamma ray bursts and black holes. They have now been created in the laboratory in a controlled fashion by irradiating a gold target with an intense picosecond duration laser pulse. About 10{sup 11} MeV positrons are emitted from the rear surface of the target in a 15 to 22-degree cone for a duration comparable to the laser pulse. These positron jets are quasi-monoenergetic (E/{delta}E {approx} 5) with peak energies controllable from 3-19 MeV. They have temperatures from 1-4 MeV in the beam frame in both the longitudinal and transverse directions. Positron production has been studied extensively in recent decades at low energies (sub-MeV) in areas related to surface science, positron emission tomography, basic antimatter science such as antihydrogen experiments, Bose-Einstein condensed positronium, and basic plasma physics. However, the experimental tools to produce very high temperature positrons and high-flux positron jets needed to simulate astrophysical positron conditions have so far been absent. The MeV temperature jets of positrons and electrons produced in our experiments offer a first step to evaluate the physics models used to explain some of the most energetic phenomena in the universe.

  13. Creating experimental color harmony map (United States)

    Chamaret, Christel; Urban, Fabrice; Lepinel, Josselin


    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.

  14. The proteolysis adaptor, NblA, binds to the N-terminus of β-phycocyanin: Implications for the mechanism of phycobilisome degradation. (United States)

    Nguyen, Amelia Y; Bricker, William P; Zhang, Hao; Weisz, Daniel A; Gross, Michael L; Pakrasi, Himadri B


    Phycobilisome (PBS) complexes are massive light-harvesting apparati in cyanobacteria that capture and funnel light energy to the photosystem. PBS complexes are dynamically degraded during nutrient deprivation, which causes severe chlorosis, and resynthesized during nutrient repletion. PBS degradation occurs rapidly after nutrient step down, and is specifically triggered by non-bleaching protein A (NblA), a small proteolysis adaptor that facilitates interactions between a Clp chaperone and phycobiliproteins. Little is known about the mode of action of NblA during PBS degradation. In this study, we used chemical cross-linking coupled with LC-MS/MS to investigate the interactions between NblA and phycobiliproteins. An isotopically coded BS(3) cross-linker captured a protein interaction between NblA and β-phycocyanin (PC). LC-MS/MS analysis identified the amino acid residues participating in the binding reaction, and demonstrated that K(52) in NblA is cross-linked to T(2) in β-PC. These results were modeled onto the existing crystal structures of NblA and PC by protein docking simulations. Our data indicate that the C-terminus of NblA fits in an open groove of β-PC, a region located inside the central hollow cavity of a PC rod. NblA may mediate PBS degradation by disrupting the structural integrity of the PC rod from within the rod. In addition, M(1)-K(44) and M(1)-K(52) cross-links between the N-terminus of NblA and the C-terminus of NblA are consistent with the NblA crystal structure, confirming that the purified NblA is structurally and biologically relevant. These findings provide direct evidence that NblA physically interacts with β-PC.

  15. Alternative Splicing at C Terminus of CaV1.4 Calcium Channel Modulates Calcium-dependent Inactivation, Activation Potential, and Current Density (United States)

    Tan, Gregory Ming Yeong; Yu, Dejie; Wang, Juejin; Soong, Tuck Wah


    The CaV1.4 voltage-gated calcium channel is predominantly expressed in the retina, and mutations to this channel have been associated with human congenital stationary night blindness type-2. The L-type CaV1.4 channel displays distinct properties such as absence of calcium-dependent inactivation (CDI) and slow voltage-dependent inactivation (VDI) due to the presence of an autoinhibitory domain (inhibitor of CDI) in the distal C terminus. We hypothesized that native CaV1.4 is subjected to extensive alternative splicing, much like the other voltage-gated calcium channels, and employed the transcript scanning method to identify alternatively spliced exons within the CaV1.4 transcripts isolated from the human retina. In total, we identified 19 alternative splice variations, of which 16 variations have not been previously reported. Characterization of the C terminus alternatively spliced exons using whole-cell patch clamp electrophysiology revealed a splice variant that exhibits robust CDI. This splice variant arose from the splicing of a novel alternate exon (43*) that can be found in 13.6% of the full-length transcripts screened. Inclusion of exon 43* inserts a stop codon that truncates half the C terminus. The CaV1.4 43* channel exhibited robust CDI, a larger current density, a hyperpolarized shift in activation potential by ∼10 mV, and a slower VDI. Through deletional experiments, we showed that the inhibitor of CDI was responsible for modulating channel activation and VDI, in addition to CDI. Calcium currents in the photoreceptors were observed to exhibit CDI and are more negatively activated as compared with currents elicited from heterologously expressed full-length CaV1.4. Naturally occurring alternative splice variants may in part contribute to the properties of the native CaV1.4 channels. PMID:22069316

  16. Interactions between the C-terminus of Kv1.5 and Kvβ regulate pyridine nucleotide-dependent changes in channel gating (United States)

    Tipparaju, Srinivas M.; Li, Xiao-Ping; Kilfoil, Peter J.; Xue, Bin; Uversky, Vladimir N.; Bhatnagar, Aruni; Barski, Oleg A.


    Voltage-gated potassium (Kv) channels are tetrameric assemblies of transmembrane Kv proteins with cytosolic N- and C-termini. The N-terminal domain of Kv1 proteins binds to β-subunits, but the role of the C-terminus is less clear. Therefore, we studied the role of the C-terminus in regulating Kv1.5 channel and its interactions with Kvβ-subunits. When expressed in COS-7 cells, deletion of the C-terminal domain of Kv1.5 did not affect channel gating or kinetics. Co-expression of Kv1.5 with Kvβ3 increased current inactivation, whereas Kvβ2 caused a hyperpolarizing shift in the voltage-dependence of current activation. Inclusion of NADPH in the patch pipette solution accelerated the inactivation of Kv1.5-Kvβ3 currents. In contrast, NADP+ decreased the rate and the extent of Kvβ3-induced inactivation and reversed the hyperpolarizing shift in the voltage-dependence of activation induced by Kvβ2. Currents generated by Kv1.5ΔC+Kvβ3 or Kv1.5ΔC+Kvβ2 complexes did not respond to changes in intracellular pyridine nucleotide concentration, indicating that the C-terminus is required for pyridine nucleotide-dependent interactions between Kvβ and Kv1.5. A glutathione-S-transferase (GST) fusion protein containing the C-terminal peptide of Kv1.5 did not bind to apoKvβ2, but displayed higher affinity for Kvβ2:NADPH than Kvβ2:NADP+. The GST fusion protein also precipitated Kvβ proteins from mouse brain lysates. Pull-down experiments, structural analysis and electrophysiological data indicated that a specific region of the C-terminus (Arg543-Val583) is required for Kvβ binding. These results suggest that the C-terminal domain of Kv1.5 interacts with β-subunits and that this interaction is essential for the differential regulation of Kv currents by oxidized and reduced nucleotides. PMID:22426702

  17. India creates social marketing organization. (United States)


    India, in a major policy shift toward reversible birth controls methods, will form a new organization to promote private sector contraceptive sales. The government, through a recently signed agreement with the Agency for International Development (AID), plans to establish a private nonprofit Contraceptive Marketing Organization (CMO) in fiscal year 1984. This momentous move marks a full circle return to a 1969 proposal by AID and Ford Foundation consultants. Funded at about $500 million over a 7 year period, the CMO will function as a semi-autonomous entity run by a board of governors representing government and such public and public sectors as health, communications, management, manufacturing, marketing, advertising, and market research. According to the agreement called the India Family Planning Communications and Marketing Plan, the CMO's activities will cover procurement and distribution of condoms, oral contraceptives (OCs), and other yet to be determined contraceptive methods. Of the $500 million in funds, the government of India has pledged 2/3, AID roughly $50 million in grants and loans, with the balance expected from such sources as the UN Fund for Population Activities. The CMO's goal is a marked increase in contraceptive use by married couples of reproductive age from the current 6% rate to 20% by 1990. As of 1982, India has 122 million such couples, with 1% purchasing commercial products, 2% buying Nirodh Marketing Program condoms and 3% relying on free government contraceptives. Besides creating the CMO, the India/AID pact outlines intensified public sector family planning promotions and activities. Some Indian health experts believe the government's decision to expand social marketing's role rests with a significant decade long decline in the popularity of such permanent birth control measures as vasectomy and tubal ligation.

  18. Creating Shared Value by Combatting Corruption

    National Research Council Canada - National Science Library

    Philip M Nichols


      Creating Shared Value The creating shared value strategy is similar to corporate social responsibility and to social impact in that it focusses on the intersection between business and the rest of society...

  19. Fusion of green fluorescent protein to the C-terminus of granulysin alters its intracellular localization in comparison to the native molecule

    Directory of Open Access Journals (Sweden)

    Ziegler Steven F


    Full Text Available Abstract The engineering of green fluorescent protein (GFP fusion constructs in order to visibly tag a protein of interest has become a commonly used cell biology technique. Although caveats to this approach are obvious, literature reports in which the chimeric molecule behaves differently than the native molecule are scant. This brief report describes one such case. Granulysin, a small lytic and antimicrobial protein produced by cytotoxic lymphocytes, traffics to the regulated secretory system and is subsequently released from cells upon proper stimulus. In an attempt to elucidate mechanisms by which it accumulates in and is released from cytolytic granules, GFP was fused to the C-terminus of granulysin and expressed in an NK cell line. A control construct expressing the native protein was similarly expressed. The data demonstrate that, while the fusion protein is expressed and secreted, its subcellular localization is altered in comparison to native granulysin. Thus, the addition of GFP to the C-terminus of granulysin obscures the signal(s that cytotoxic lymphocytes use to sort it to the regulated secretory pathway despite its normal biosynthesis and secretion. This example is offered as a cautionary account for other researchers who contemplate using this technology.

  20. Procerain B, a cysteine protease from Calotropis procera, requires N-terminus pro-region for activity: cDNA cloning and expression with pro-sequence. (United States)

    Nandana, Vidhyadhar; Singh, Sushant; Singh, Abhay Narayan; Dubey, Vikash Kumar


    We have previously reported isolation and characterization of a novel plant cysteine protease, Procerain B, from the latex of Calotropis procera. Our initial attempts for active recombinant Procerain B in Escherichiacoli expression system was not successful. The reason for inactive enzyme production was attributed to the absence of 5' pro-region in the Procerain B cDNA that may be involved in proper folding and production of mature active protein. The current manuscript reports the cloning of full length Procerain B for the production of the active protein. The complete cDNA sequence of Procerain B with pro-region sequence was obtained by using RNA ligase mediated rapid amplification of 5' cDNA ends (RLM-RACE). The N-terminus pro-sequence region consists of 127 amino acids and characterized as the member of inhibitory I29 family. Further the three dimensional structure of full length Procerain B was modelled by homology modelling using X-ray crystal structure of procaricain (PDB ID: 1PCI). N-terminus pro-sequence of full length Procerain B runs along the active site cleft. Full length Procerain B was expressed in prokaryotic system and activated in vitro at pH 4.0. This is the first study reporting the production of active recombinant cysteine protease from C.procera. Copyright © 2014 Elsevier Inc. All rights reserved.

  1. Allosteric coupling between proximal C-terminus and selectivity filter is facilitated by the movement of transmembrane segment 4 in TREK-2 channel. (United States)

    Zhuo, Ren-Gong; Peng, Peng; Liu, Xiao-Yan; Yan, Hai-Tao; Xu, Jiang-Ping; Zheng, Jian-Quan; Wei, Xiao-Li; Ma, Xiao-Yun


    TREK-2, a member of two-pore-domain potassium channel family, regulates cellular excitability in response to diverse stimuli. However, how such stimuli control channel function remains unclear. Here, by characterizing the responses of cytosolic proximal C-terminus deletant (ΔpCt) and transmembrane segment 4 (M4)-glycine hinge mutant (G312A) to 2-Aminoethoxydiphenyl borate (2-APB), an activator of TREK-2, we show that the transduction initiated from pCt domain is allosterically coupled with the conformation of selectivity filter (SF) via the movements of M4, without depending on the original status of SF. Moreover, ΔpCt and G312A also exhibited blunted responses to extracellular alkalization, a model to induce SF conformational transition. These results suggest that the coupling between pCt domain and SF is bidirectional, and M4 movements are involved in both processes. Further mechanistic exploration reveals that the function of Phe316, a residue close to the C-terminus of M4, is associated with such communications. However, unlike TREK-2, M4-hinge of TREK-1 only controls the transmission from pCt to SF, rather than SF conformational changes triggered by pHo changes. Together, our findings uncover the unique gating properties of TREK-2, and elucidate the mechanisms for how the extracellular and intracellular stimuli harness the pore gating allosterically.

  2. Dual role of φ29 DNA polymerase Lys529 in stabilisation of the DNA priming-terminus and the terminal protein-priming residue at the polymerisation site.

    Directory of Open Access Journals (Sweden)

    Alicia del Prado

    Full Text Available Resolution of the crystallographic structure of φ29 DNA polymerase binary and ternary complexes showed that residue Lys529, located at the C-terminus of the palm subdomain, establishes contacts with the 3' terminal phosphodiester bond. In this paper, site-directed mutants at this Lys residue were used to analyse its functional importance for the synthetic activities of φ29 DNA polymerase, an enzyme that starts linear φ29 DNA replication using a terminal protein (TP as primer. Our results show that single replacement of φ29 DNA polymerase residue Lys529 by Ala or Glu decreases the stabilisation of the primer-terminus at the polymerisation active site, impairing both the insertion of the incoming nucleotide when DNA and TP are used as primers and the translocation step required for the next incoming nucleotide incorporation. In addition, combination of the DNA polymerase mutants with a TP derivative at residue Glu233, neighbour to the priming residue Ser232, leads us to infer a direct contact between Lys529 and Glu233 for initiation of TP-DNA replication. Altogether, the results are compatible with a sequential binding of φ29 DNA polymerase residue Lys529 with TP and DNA during replication of TP-DNA.

  3. A C-terminal Hydrophobic, Solvent-protected Core and a Flexible N-terminus are Potentially Required for Human Papillomavirus 18 E7 Protein Functionality

    Energy Technology Data Exchange (ETDEWEB)

    Liu, S.; Tian, Y; Greenaway, F; Sun, M


    The oncogenic potential of the high-risk human papillomavirus (HPV) relies on the expression of genes specifying the E7 and E6 proteins. To investigate further the variation in oligomeric structure that has been reported for different E7 proteins, an HPV-18 E7 cloned from a Hispanic woman with cervical intraepithelial neoplasia was purified to homogeneity most probably as a stable monomeric protein in aqueous solution. We determined that one zinc ion is present per HPV-18 E7 monomer by amino acid and inductively coupled plasma-atomic emission spectroscopy analysis. Intrinsic fluorescence and circular dichroism spectroscopic results indicate that the zinc ion is important for the correct folding and thermal stability of HPV-18 E7. Hydroxyl radical mediated protein footprinting coupled to mass spectrometry and other biochemical and biophysical data indicate that near the C-terminus, the four cysteines of the two Cys-X{sub 2}-Cys motifs that are coordinated to the zinc ion form a solvent inaccessible core. The N-terminal LXCXE pRb binding motif region is hydroxyl radical accessible and conformationally flexible. Both factors, the relative flexibility of the pRb binding motif at the N-terminus and the C-terminal metal-binding hydrophobic solvent-protected core, combine together and facilitate the biological functions of HPV-18 E7.


    Directory of Open Access Journals (Sweden)

    Gusti Ayu Yuniati Kencana


    Full Text Available The information on pathogenicity and adaptation factors of avian influenza virus (AIV in mammalsis very inportant in an effort to reduce the risk of avian influenza (AI pandemic in the future. Polymerasegene complex appears to be the major factors for adaptation of AIV to certain animal species. A preliminarystudy on role of non-coding region (NCR and amino-terminus of polymerase-basic 2 (PB2 is presented.Purified viral RNA of AIV isolated from chicken, duck, pig, and quail of Bali and Yogyakarta was reversetranscribed into cDNA and amplified using reverse transcriptase-polymerase chain reaction (RT-PCRusing PB-2 universal forward primer and specifically designed backward primer. The result showed thatall AIV’s H5N1 isolated from chicken, duck, quail, and pig, posed PB2 amino-terminus typical for IndonesianAIV H5N1. However, polymorphic amino acids of the protein fragment did not show any species specificmotive, with the exception of the pig isolate Sw/Tabanan/2006 which had specific substitution of D16E,H17Q, M40I, and H124Y.

  5. Gene expression changes in serotonin, GABA-A receptors, neuropeptides and ion channels in the dorsal raphe nucleus of adolescent alcohol-preferring (P) rats following binge-like alcohol drinking. (United States)

    McClintick, Jeanette N; McBride, William J; Bell, Richard L; Ding, Zheng-Ming; Liu, Yunlong; Xuei, Xiaoling; Edenberg, Howard J


    Alcohol binge-drinking during adolescence is a serious public health concern with long-term consequences. We used RNA sequencing to assess the effects of excessive adolescent ethanol binge-drinking on gene expression in the dorsal raphe nucleus (DRN) of alcohol preferring (P) rats. Repeated binges across adolescence (three 1h sessions across the dark-cycle per day, 5 days per week for 3 weeks starting at 28 days of age; ethanol intakes of 2.5-3 g/kg/session) significantly altered the expression of approximately one-third of the detected genes. Multiple neurotransmitter systems were altered, with the largest changes in the serotonin system (21 of 23 serotonin-related genes showed decreased expression) and GABA-A receptors (8 decreased and 2 increased). Multiple neuropeptide systems were also altered, with changes in the neuropeptide Y and corticotropin-releasing hormone systems similar to those associated with increased drinking and decreased resistance to stress. There was increased expression of 21 of 32 genes for potassium channels. Expression of downstream targets of CREB signaling was increased. There were also changes in expression of genes involved in inflammatory processes, axonal guidance, growth factors, transcription factors, and several intracellular signaling pathways. These widespread changes indicate that excessive binge drinking during adolescence alters the functioning of the DRN and likely its modulation of many regions of the central nervous system, including the mesocorticolimbic system. Copyright © 2014 Elsevier Inc. All rights reserved.

  6. 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


    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

  7. Creating the Environment for Continuous Competition (United States)


    The Limits of Competition in Defense Acquisition Defense Acquisition University Research Symposium, September 2012 CREATING THE ENVIRONMENT FOR...3. DATES COVERED 00-00-2012 to 00-00-2012 4. TITLE AND SUBTITLE Creating the Environment for Continuous Competition 5a. CONTRACT NUMBER 5b...Information Technology (IT) Box A Primer. Retrieved from http: Creating the Environment for Continuous

  8. Basis of recognition between the NarJ chaperone and the N-terminus of the NarG subunit from Escherichia coli nitrate reductase. (United States)

    Zakian, Silva; Lafitte, Daniel; Vergnes, Alexandra; Pimentel, Cyril; Sebban-Kreuzer, Corinne; Toci, René; Claude, Jean-Baptiste; Guerlesquin, Françoise; Magalon, Axel


    A novel class of molecular chaperones co-ordinates the assembly and targeting of complex metalloproteins by binding to an amino-terminal peptide of the cognate substrate. We have previously shown that the NarJ chaperone interacts with the N-terminus of the NarG subunit coming from the nitrate reductase complex, NarGHI. In the present study, NMR structural analysis revealed that the NarG(1-15) peptide adopts an alpha-helical conformation in solution. Moreover, NarJ recognizes and binds the helical NarG(1-15) peptide mostly via hydrophobic interactions as deduced from isothermal titration calorimetry analysis. NMR and differential scanning calorimetry analysis revealed a modification of NarJ conformation during complex formation with the NarG(1-15) peptide. Isothermal titration calorimetry and BIAcore experiments support a model whereby the protonated state of the chaperone controls the time dependence of peptide interaction.

  9. The effect of structural differences in the reducing terminus of sugars on the binding affinity of carbohydrates and proteins analyzed using photoaffinity labeling. (United States)

    Ohtsuka, Isao; Sadakane, Yutaka; Higuchi, Mari; Hada, Noriyasu; Hada, Junko; Kakiuchi, Nobuko; Sakushima, Akiyo


    Because carbohydrates and proteins bind with such low affinity, the nature of their interactions is not clear. Photoaffinity labeling with diazirin groups is useful for elucidating the roles of carbohydrates in these binding processes. However, when carbohydrate probes are synthesized according to this conventional method, the reducing terminus of the sugar is opened to provide an acyclic structure. Because greater elucidation of carbohydrate-protein interactions requires a closed-ring carbohydrate in addition to the photoreactive group, we synthesized new molecular tools. The carbohydrate ligands were synthesized in three steps (glycosylation with allyl alcohol, deprotection, and ozonolysis). Specific binding proteins for carbohydrate ligands were obtained by photoaffinity labeling. Closed ring-type carbohydrate ligands, in which the reducing sugar is closed, bound to lectins more strongly than open ring-type sugars. Carbohydrate to protein binding was observed using AFM. Copyright © 2010 Elsevier Ltd. All rights reserved.

  10. A Conserved Tripeptide Sequence at the C Terminus of the Poxvirus DNA Processivity Factor D4 Is Essential for Protein Integrity and Function. (United States)

    Nuth, Manunya; Guan, Hancheng; Ricciardi, Robert P


    Vaccinia virus (VACV) is a poxvirus, and the VACV D4 protein serves both as a uracil-DNA glycosylase and as an essential component required for processive DNA synthesis. The VACV A20 protein has no known catalytic function itself but associates with D4 to form the D4-A20 heterodimer that functions as the poxvirus DNA processivity factor. The heterodimer enables the DNA polymerase to efficiently synthesize extended strands of DNA. Upon characterizing the interaction between D4 and A20, we observed that the C terminus of D4 is susceptible to perturbation. Further analysis demonstrated that a conserved hexapeptide stretch at the extreme C terminus of D4 is essential for maintaining protein integrity, as assessed by its requirement for the production of soluble recombinant protein that is functional in processive DNA synthesis. From the known crystal structures of D4, the C-terminal hexapeptide is shown to make intramolecular contact with residues spanning the inner core of the protein. Our mutational analysis revealed that a tripeptide motif ((215)GFI(217)) within the hexapeptide comprises apparent residues necessary for the contact. Prediction of protein disorder identified the hexapeptide and several regions upstream of Gly(215) that comprise residues of the interface surfaces of the D4-A20 heterodimer. Our study suggests that (215)GFI(217) anchors these potentially dynamic upstream regions of the protein to maintain protein integrity. Unlike uracil-DNA glycosylases from diverse sources, where the C termini are disordered and do not form comparable intramolecular contacts, this feature may be unique to orthopoxviruses. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

  11. 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)


    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.

  12. Moesin-ezrin-radixin-like protein (merlin) mediates protein interacting with the carboxyl terminus-1 (PICT-1)-induced growth inhibition of glioblastoma cells in the nucleus. (United States)

    Chen, Hongbo; Mei, Lin; Zhou, Lanzhen; Zhang, Xudong; Guo, Caiping; Li, Junchang; Wang, Huixia; Zhu, Yongqiang; Zheng, Yi; Huang, Laiqiang


    Moesin-ezrin-radixin-like protein (merlin) has long been considered a unique tumour suppressor that inhibits mitogenic signalling only at the membrane-cytoskeleton interface. However, the nucleocytoplasmic shuttling of merlin in a cell cycle-dependent manner has recently been observed, indicating that merlin may also exert its tumour-suppressive activity by interacting with specific nuclear protein partners. We have identified protein interacting with carboxyl terminus 1 (PICT-1) as a novel merlin-binding partner. Although the detailed mechanisms are not fully understood, several lines of evidence have previously implicated PICT-1 as a candidate tumour suppressor, including its phosphatase and tensin homolog deleted on chromosome 10 (PTEN)-dependent growth-suppression and cell-killing activities. We show here that PICT-1 is localised to the nucleolus, and Ser518-dephosphorylated merlin (the growth-inhibitory form of merlin) can interact with PICT-1 in the nucleolus. Ectopic expression of PICT-1, both in PTEN-positive HeLa cells and in PTEN-deficient U251 cells, effectively represses cyclin D1 expression, arrests the cell cycle at G0/G1, and promotes cell apoptosis. PICT-1 (1-356), a carboxyl-terminus truncated mutant that has lost the ability to bind merlin, has a markedly reduced inhibitory effect on the cell cycle and proliferation. Knockdown of merlin expression by siRNA attenuates the inhibitory effects induced by PICT-1 over-expression. We propose that merlin mediates PICT-1-induced growth inhibition by translocating to the nucleolus and binding PICT-1. Copyright © 2010 Elsevier Ltd. All rights reserved.

  13. Positively charged amino acids at the SNAP-25 C terminus determine fusion rates, fusion pore properties, and energetics of tight SNARE complex zippering. (United States)

    Fang, Qinghua; Zhao, Ying; Herbst, Adam Drew; Kim, Brian N; Lindau, Manfred


    SNAP-25 is a Q-SNARE protein mediating exocytosis of neurosecretory vesicles including chromaffin granules. Previous results with a SNAP-25 construct lacking the nine C terminal residues (SNAP-25Δ9) showed changed fusion pore properties (Fang et al., 2008), suggesting a model for fusion pore mechanics that couple C terminal zipping of the SNARE complex to the opening of the fusion pore. The deleted fragment contains the positively charged residues R198 and K201, adjacent to layers 7 and 8 of the SNARE complex. To determine how fusion pore conductance and dynamics depend on these residues, single exocytotic events in bovine chromaffin cells expressing R198Q, R198E, K201Q, or K201E mutants were investigated by carbon fiber amperometry and cell-attached patch capacitance measurements. Coarse grain molecular dynamics simulations revealed spontaneous transitions between a loose and tightly zippered state at the SNARE complex C terminus. The SNAP-25 K201Q mutant showed no changes compared with SNAP-25 wild-type. However, K201E, R198Q, and R198E displayed reduced release frequencies, slower release kinetics, and prolonged fusion pore duration that were correlated with reduced probability to engage in the tightly zippered state. The results show that the positively charged amino acids at the SNAP-25 C terminus promote tight SNARE complex zippering and are required for high release frequency and rapid release in individual fusion events. Copyright © 2015 the authors 0270-6474/15/353230-10$15.00/0.

  14. Off-Target V(DJ Recombination Drives Lymphomagenesis and Is Escalated by Loss of the Rag2 C Terminus

    Directory of Open Access Journals (Sweden)

    Martina Mijušković


    Full Text Available Genome-wide analysis of thymic lymphomas from Tp53−/− mice with wild-type or C-terminally truncated Rag2 revealed numerous off-target, RAG-mediated DNA rearrangements. A significantly higher fraction of these errors mutated known and suspected oncogenes/tumor suppressor genes than did sporadic rearrangements (p < 0.0001. This tractable mouse model recapitulates recent findings in human pre-B ALL and allows comparison of wild-type and mutant RAG2. Recurrent, RAG-mediated deletions affected Notch1, Pten, Ikzf1, Jak1, Phlda1, Trat1, and Agpat9. Rag2 truncation substantially increased the frequency of off-target V(DJ recombination. The data suggest that interactions between Rag2 and a specific chromatin modification, H3K4me3, support V(DJ recombination fidelity. Oncogenic effects of off-target rearrangements created by this highly regulated recombinase may need to be considered in design of site-specific nucleases engineered for genome modification.

  15. Injections of the of the α1-adrenoceptor antagonist prazosin into the median raphe nucleus increase food intake and Fos expression in orexin neurons of free-feeding rats. (United States)

    Silva, Eduardo Simão da; Flores, Rafael Appel; Ribas, Anderson Savaris; Taschetto, Ana Paula; Faria, Moacir Serralvo; Lima, Leandro Bueno; Metzger, Martin; Donato, José; Paschoalini, Marta Aparecida


    Previously, we showed that the blockade of α1-adrenoreceptors in the median raphe nucleus (MnR) increased food intake in free-feeding rats, indicating that adrenergic mechanisms in the MnR participate in the regulation of food intake. However, the impact of such a pharmacological manipulation on other neural circuits related to food intake remains unknown. In the current study, we sought to identify forebrain regions which are responsive to α1-adrenergic receptor blockade and presumably involved in the modulation of the feeding response. For this purpose, we examined the induction of c-Fos immunoreactivity in forebrain structures following injections of the α1-adrenoceptor antagonist prazosin into the MnR of free-feeding rats. To determine the chemical identity of hypothalamic c-Fos-positive cells, we then conducted double-label immunohistochemistry for Fos/orexin (OX) or Fos/melanin-concentrating hormone (MCH). Finally, we combined anterograde tracing from the MnR with immunohistochemical detection of orexin. Prazosin injections into the MnR significantly increased food intake. The ingestive response was accompanied by an increase in Fos expression in the basolateral amygdala (BLA) and lateral hypothalamic area (LHA). In the LHA, Fos expression occurred in neurons expressing OX, but not MCH. Combined anterograde tracing experiments revealed that LHA OX neurons are prominently targeted by MnR axons. These findings suggest that intra-MnR injection of prazosin, via activation of orexinergic neurons in the LHA and non-orexinergic neurons in the BLA, evoked a motivational response toward food intake. Copyright © 2017 Elsevier B.V. All rights reserved.

  16. Process to create simulated lunar agglutinate particles (United States)

    Gustafson, Robert J. (Inventor); Gustafson, Marty A. (Inventor); White, Brant C. (Inventor)


    A method of creating simulated agglutinate particles by applying a heat source sufficient to partially melt a raw material is provided. The raw material is preferably any lunar soil simulant, crushed mineral, mixture of crushed minerals, or similar material, and the heat source creates localized heating of the raw material.

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

    African Journals Online (AJOL)

    Thus, unless there is effort to create self employment, this can galvanize unexpected revolution whose consequences will be very grave. The reading public will have to apply the advice provided in this article to create self employment. Key Words: e-commerce, online transaction, broadband stimulus, unemployment.

  18. Creating Safe Spaces for Music Learning (United States)

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


    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,…

  19. Creating mobile apps with Appcelerator Titanium

    CERN Document Server

    Brousseau, Christian


    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

  20. Creating Web Sites The Missing Manual

    CERN Document Server

    MacDonald, Matthew


    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

  1. Creating Healthy Environments For Youth Athletes (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.

  2. More Sci- than Fi, Physicists Create Antimatter

    CERN Document Server

    Overbye, Dennis


    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)

  3. How to Create a Personal Health Record (United States)

    ... currently available to help you create your own personal health record ( PHR ). PHR s are an inevitable and critical step in the evolution of health information management ( HIM ). The book, “The ...

  4. A summary of the CMS Create event

    CERN Multimedia

    CERN. Geneva; GASTAL, Martin


    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!

  5. Taking medicine at home - create a routine (United States)

    ... page: // 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, ...

  6. How to Create a Reference from XML (United States)

    US Fish and Wildlife Service, Department of the Interior — This module walks through the process of manually creating a ‘Geospatial Data’ record from an eXtensible Markup Language (XML) metadata file. It assumes that you...

  7. Creating competence: perspectives and practices in organizations

    NARCIS (Netherlands)

    Mulder, Martin


    Creating competence has become a major issue in organizations. Various authors contend that competency management has the potential of integrating organizational strategy, human-resource instruments, and human-resource development; that competency development can lead to performance improvement; and

  8. Creating Math Videos: Comparing Platforms and Software (United States)

    Abbasian, Reza O.; Sieben, John T.


    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…

  9. Co-creating meaning through Artful Inquiry

    DEFF Research Database (Denmark)

    Darsø, Lotte


    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...... of leadership icons as well as co-creating with tangible materials can give rise to new meaning and transformational learning....

  10. ISO 55000: Creating an asset management system. (United States)

    Bradley, Chris; Main, Kevin


    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.

  11. A leucine residue in the C terminus of human parainfluenza virus type 3 matrix protein is essential for efficient virus-like particle and virion release. (United States)

    Zhang, Guangyuan; Zhang, Shengwei; Ding, Binbin; Yang, Xiaodan; Chen, Longyun; Yan, Qin; Jiang, Yanliang; Zhong, Yi; Chen, Mingzhou


    Paramyxovirus particles, like other enveloped virus particles, are formed by budding from membranes of infected cells, and matrix (M) proteins are critical for this process. To identify the M protein important for this process, we have characterized the budding of the human parainfluenza virus type 3 (HPIV3) M protein. Our results showed that expression of the HPIV3 M protein alone is sufficient to initiate the release of virus-like particles (VLPs). Electron microscopy analysis confirmed that VLPs are morphologically similar to HPIV3 virions. We identified a leucine (L302) residue within the C terminus of the HPIV3 M protein that is critical for M protein-mediated VLP production by regulating the ubiquitination of the M protein. When L302 was mutated into A302, ubiquitination of M protein was defective, the release of VLPs was abolished, and the membrane binding and budding abilities of M protein were greatly weakened, but the ML302A mutant retained oligomerization activity and had a dominant negative effect on M protein-mediated VLP production. Furthermore, treatment with a proteasome inhibitor also inhibited M protein-mediated VLP production and viral budding. Finally, recombinant HPIV3 containing the M(L302A) mutant could not be rescued. These results suggest that L302 acts as a critical regulating signal for the ubiquitination of the HPIV3 M protein and virion release. Human parainfluenza virus type 3 (HPIV3) is an enveloped virus with a nonsegmented negative-strand RNA genome. It can cause severe respiratory tract diseases, such as bronchiolitis, pneumonia, and croup in infants and young children. However, no valid antiviral therapy or vaccine is currently available. Thus, further elucidation of its assembly and budding will be helpful in the development of novel therapeutic approaches. Here, we show that a leucine residue (L302) located at the C terminus of the HPIV3 M protein is essential for efficient production of virus-like particles (VLPs). Furthermore

  12. Role of the N-terminus in determining metal-specific responses in the E. coli Ni- and Co-responsive metalloregulator, RcnR. (United States)

    Higgins, Khadine A; Chivers, Peter T; Maroney, Michael J


    RcnR (resistance to cobalt and nickel regulator) is a 40-kDa homotetrameric protein and metalloregulator that controls the transcription of the Co(II) and Ni(II) exporter, RcnAB, by binding to DNA as an apoprotein and releasing DNA in response to specifically binding Co(II) and Ni(II) ions. Using X-ray absorption spectroscopy (XAS) to examine the structure of metals bound and lacZ reporter assays of the transcription of RcnA in response to metal binding, in WT and mutant proteins, the roles of coordination number, ligand selection, and residues in the N-terminus of the protein were examined as determinants in metal ion recognition. The studies show that the cognate metal ions, Co(II) and Ni(II), which bind in (N/O)(5)S six-coordinate sites, are distinguished from non-cognate metal ions (Cu(I) and Zn(II)), which bind only three protein ligands and one anion from the buffer, by coordination number and ligand selection. Using mutations of residues near the N-terminus, the N-terminal amine is shown to be a ligand of the cognate metal ions that is missing in the complexes with non-cognate metal ions. The side chain of His3 is also shown to play an important role in distinguishing metal ions. The imidazole group is shown to be a ligand in the Co(II) RcnR complex, but not in the Zn(II) complex. Further, His3 does not appear to bind to Ni(II), providing a structural basis for the differential regulation of RcnAB by the two cognate ions. The Zn(II) complexes change coordination number in response to the residue in position three. In H3C-RcnR, the Zn(II) complex is five-coordinate, and in H3E-RcnR the Zn(II) ion is bound to six protein ligands. The metric parameters of this unusual Zn(II) structure resemble those of the WT-Ni(II) complex, and the mutant protein is able to regulate expression of RcnAB in response to binding the non-cognate ion. The results are discussed within a protein allosteric model for gene regulation by metalloregulators.

  13. hERG 1a LQT2 C-terminus truncation mutants display hERG 1b-dependent dominant negative mechanisms. (United States)

    Puckerin, Akil; Aromolaran, Kelly A; Chang, Donald D; Zukin, R Suzanne; Colecraft, Henry M; Boutjdir, Mohamed; Aromolaran, Ademuyiwa S


    The human ether-à-go-go-related gene (hERG 1a) potassium channel is critical for cardiac repolarization. hERG 1b, another variant subunit, co-assembles with hERG 1a, modulates channel biophysical properties and plays an important role in repolarization. Mutations of hERG 1a lead to type 2 long QT syndrome (LQT2), and increased risk for fatal arrhythmias. The functional consequences of these mutations in the presence of hERG 1b are not known. To investigate whether hERG 1a mutants exert dominant negative gating and trafficking defects when co-expressed with hERG 1b. Electrophysiology, co-immunoprecipitation, and fluorescence resonance energy transfer (FRET) experiments in HEK293 cells and guinea pig cardiomyocytes were used to assess the mutants on gating and trafficking. Mutations of 1a-G965X and 1a-R1014X, relevant to gating and trafficking were introduced in the C-terminus region. The hERG 1a mutants when expressed alone did not result in decreased current amplitude. Compared to wild-type hERG 1a currents, 1a-G965X currents were significantly larger, whereas those produced by the 1a-R1014X mutant were similar in magnitude. Only when co-expressed with wild-type hERG 1a and 1b did a mutant phenotype emerge, with a marked reduction in surface expression, current amplitude, and a corresponding positive shift in the V1/2 of the activation curve. Co-immunoprecipitation and FRET assays confirmed association of mutant and wild-type subunits. Heterologously expressed hERG 1a C-terminus truncation mutants, exert a dominant negative gating and trafficking effect only when co-expressed with hERG 1b. These findings may have potentially profound implications for LQT2 therapy. Copyright © 2016 Heart Rhythm Society. All rights reserved.

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

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


    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

  15. Modifying the N-terminus of polyamides: PyImPyIm has improved sequence specificity over f-ImPyIm. (United States)

    Brown, Toni; Mackay, Hilary; Turlington, Mark; Sutterfield, Arden; Smith, Traci; Sielaff, Alan; Westrate, Laura; Bruce, Chrystal; Kluza, Jerome; O'Hare, Caroline; Nguyen, Binh; Wilson, W David; Hartley, John A; Lee, Moses


    Seven N-terminus modified derivatives of a previously published minor-groove binding polyamide (f-ImPyIm, 1) were synthesized and the biochemical and biophysical chemistry evaluated. These compounds were synthesized with the aim of attaining a higher level of sequence selectivity over f-ImPyIm (1), a previously published strong minor-groove binder. Two compounds possessing a furan or a benzofuran moiety at the N-terminus showed a footprint of 0.5microM at the cognate ACGCGT site (determined by DNase I footprinting); however, the specificity of these compounds was not improved. In contrast, PyImPyIm (4) produced a footprint of 0.5microM but showed a superior specificity using the same technique. When evaluated by thermal melting experiments and circular dichroism using ACGCGT and the non-cognate AAATTT sequence, all compounds were shown to bind in the minor-groove of DNA and stabilize the cognate sequence much better than the non-cognate (except for the non-amido-compound that did not bind either sequence, as expected). PyImPyIm (4) was interesting as the DeltaT(m) for this compound was only 4 degrees C but the footprint was very selective. No binding was observed for this compound with a third DNA (non-cognate, ACCGGT). ITC studies on compound 4 showed exothermic binding with ACGCGT and no heat change was observed for titrating the compound to the other two DNA sequences. The heat capacity (DeltaC(p)) of the PIPI/ACGCGT complex calculated from the hydrophobic interactions and SASA calculations was comparable to the experimental value obtained from ITC (-146calmol(-1)K(-1)). SPR results provided confirmation of the sequence specificity of PyImPyIm (4), with a K(eq) value determined to be 7.1x10(6) M(-1) for the cognate sequence and no observable binding to AAATTT and ACCGGT. Molecular dynamic simulations affirmed that PyImPyIm (4) binds as a dimer in an overlapped conformation, and it fits snugly in the minor-groove of the ACGCGT oligonucleotide. PyImPyIm (4) is an

  16. 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: [Division of Biological Science, Graduate School of Science, Nagoya University (Japan); Structural Biology Research Center, Graduate School of Science, Nagoya University (Japan)


    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

  17. Adaptive Changes in the Sensitivity of the Dorsal Raphe and Hypothalamic Paraventricular Nuclei to Acute Exercise, and Hippocampal Neurogenesis May Contribute to the Antidepressant Effect of Regular Treadmill Running in Rats

    Directory of Open Access Journals (Sweden)

    Ayu Nishii


    Full Text Available Increasing clinical evidence suggests that regular physical exercise can prevent or reduce the incidence of stress-related psychiatric disorders including depressive symptoms. Antidepressant effect of regular exercise may be implicated in monoaminergic transmission including serotonergic transmission, activation of the hypothalamic-pituitary-adrenal (HPA axis, and hippocampal neurogenesis, but few general concepts regarding the optimal exercise regimen for stimulating neural mechanisms involved in antidepressant properties have been developed. Here, we examined how 4 weeks of treadmill running at different intensities (0, 15, 25 m/min, 60 min/day, 5 times/week alters neuronal activity in the dorsal raphe nucleus (DRN, which is the major source of serotonin (5-HT neurons in the central nervous system, and the hypothalamic paraventricular nucleus (PVN, in which corticotropin-releasing factor (CRF neurons initiate the activation of the HPA axis, during one session of acute treadmill running at different speeds (0, 15, 25 m/min, 30 min in male Wistar rats, using c-Fos immunohistochemistry. We also examined neurogenesis in the hippocampus using immunohistochemistry for doublecortin (DCX and assessed depressive-like behavior using the forced swim test after regular exercise for 4 weeks. In the pre-training period, acute treadmill running at low speed, but not at high speed, increased c-Fos positive nuclei in the DRN compared with the sedentary control. The number of c-Fos positive nuclei in the PVN during acute treadmill running was increased in a running speed-dependent manner. Regular exercise for 4 weeks, regardless of the training intensity, induced an enhancement of c-Fos expression in the DRN during not only low-speed but also high-speed acute running, and generally reduced c-Fos expression in the PVN during acute running compared with pre-training. Furthermore, regular treadmill running for 4 weeks enhanced DCX immunoreactivity in the

  18. Actin-ADF/cofilin rod formation in Caenorhabditis elegans muscle requires a putative F-actin binding site of ADF/cofilin at the C-terminus. (United States)

    Ono, Kanako; Ono, Shoichiro


    Under a number of stress or pathological conditions, actin and actin depolymerizing factor (ADF)/cofilin form rod-like structures that contain abnormal bundles of actin filaments that are heavily decorated with ADF/cofilin. However, the mechanism of actin rod formation and the physiological role of actin rods are not clearly understood. Here, we report that overexpression of green fluorescent protein-fused UNC-60B, a muscle-specific ADF/cofilin isoform, in Caenorhabditis elegans body wall muscle induces formation of rod-like structures. The rods contained GFP-UNC-60B, actin-interacting protein 1 (AIP1), and actin, but not other major actin-associated proteins, thus resembling actin-ADF/cofilin rods found in other organisms. However, depletion or overexpression of AIP1 did not affect formation of the actin-GFP-UNC-60B rods, suggesting that AIP1 does not play a significant role in the rod assembly. Truncation of the C-terminal tail, a putative F-actin binding site, of UNC-60B abolished induction of the rod formation, strongly suggesting that stable association of UNC-60B with F-actin, which is mediated by its C-terminus, is required for inducing actin-ADF/cofilin rods. This study suggests that C. elegans can be a new model to study functions of actin-ADF/cofilin rods. (c) 2009 Wiley-Liss, Inc.

  19. CHIP, a carboxy terminus HSP-70 interacting protein, prevents cell death induced by endoplasmic reticulum stress in the central nervous system

    Directory of Open Access Journals (Sweden)

    Felipe eCabral Miranda


    Full Text Available Endoplasmic reticulum (ER stress and protein misfolding are associated with various neurodegenerative diseases. ER stress activates Unfolded Protein Response (UPR, an adaptative response. However, severe ER stress can induce cell death. Here we show that the E3 ubiquitin ligase and co-chaperone Carboxyl Terminus HSP70/90 Interacting Protein (CHIP prevents neuron death in the hippocampus induced by severe ER stress. Organotypic hippocampal slice cultures (OHSCs were exposed to Tunicamycin, a pharmacological ER stress inducer, to trigger cell death. Overexpression of CHIP was achieved with a recombinant adeno-associated viral vector (rAAV and significantly diminished ER stress-induced cell death, as shown by analysis of propidium iodide (PI uptake, condensed chromatin, TUNEL and cleaved caspase 3 in the CA1 region of OHSCs. In addition, overexpression of CHIP prevented upregulation of both CHOP and p53 both pro-apoptotic pathways induced by ER stress. We also detected an attenuation of eIF2a phosphorylation promoted by ER stress. However, CHIP did not prevent upregulation of BiP/GRP78 induced by UPR. These data indicate that overexpression of CHIP attenuates ER-stress death response while maintain ER stress adaptative response in the central nervous system. These results indicate a neuroprotective role for CHIP upon UPR signalling. CHIP emerge as a candidate for clinical intervention in neurodegenerative diseases associated with ER stress.

  20. A Proline-Rich Domain in the Genotype 4 Hepatitis E Virus ORF3 C-Terminus Is Crucial for Downstream V105DLP108 Immunoactivity.

    Directory of Open Access Journals (Sweden)

    Heng Wang

    Full Text Available The hepatitis E virus (HEV is responsible for serious viral hepatitis worldwide. Animals are considered a reservoir of HEV, particularly pigs. While HEV infection in pigs and dogs is always asymptomatic, the virus causes high death rates in patients with pre-existing chronic liver disease and pregnant women in developing countries. HEV open reading frame 2 (ORF2 has been used as a diagnostic target to detect specific antibodies against HEV in serum samples. Recent research has additionally supported the potential utility of the ORF3 protein as a target in serum anti-HEV detection. However, the epitope distribution of ORF3 protein remains ambiguous. In the current study, we showed that continuous amino acid motif, VDLP, at the C-terminus of genotype 4 HEV ORF3 is a core sequence of the ORF3 protein epitope. Moreover, cooperative interaction with upstream elements is essential for its immunoactivity. Three proline residues (P99, P102 and P103 in the upstream proline-rich domain exerted significant effects on the immunocompetence of VDLP. ELISA results revealed that SAPPLPPVVDLP and SAPPLPPVVDLPQLGL peptides containing the identified VDLP epitope display weaker reactions with anti-HEV serum than the commercial ELISA kit. Our collective findings provide valuable information on the epitope distribution characteristics of HEV ORF3 and improve our understanding of the influence of the proline-rich domain on the immunoactivity of downstream amino acids in the C-terminal region.

  1. Adhesive Forces between A1 Domain of von Willebrand Factor and N-terminus Domain of Glycoprotein Ibα Measured by Atomic Force Microscopy. (United States)

    Tobimatsu, Hiroaki; Nishibuchi, Yuichiro; Sudo, Ryo; Goto, Shinya; Tanishita, Kazuo


    von Willebrand factor (VWF) plays an important role in the regulation of hemostasis and thrombosis formation, particularly under a high shear rate. However, the adhesive force due to the molecular interaction between VWF and glycoprotein Ibα (GPIbα) has not been fully explored. Thus, we employed atomic force microscopy to directly measure the adhesive force between VWF and GPIbα. We measured the adhesive force between VWF and GPIbα at the molecular level using an atomic force microscope (AFM). An AFM cantilever was coated with recombinant N-terminus VWF binding site of GPIbα, whereas a cover glass was coated with native VWF. The adhesive force at the molecular level was measured using an AFM. In the presence of 1 μg/mL VWF, the adhesion force was nearly 200 pN. As per the Gaussian fit analysis, the adhesive force of a single bond could have been 54 or 107 pN. Our consideration with the Gaussian fit analysis proposed that the adhesive force of a single bond could be 54 pN, which is very close to that obtained by optical tweezers (50 pN).

  2. The Carboxy Terminus of the Ligand Peptide Determines the Stability of the MHC Class I Molecule H-2Kb: A Combined Molecular Dynamics and Experimental Study.

    Directory of Open Access Journals (Sweden)

    Esam Tolba Abualrous

    Full Text Available Major histocompatibility complex (MHC class I molecules (proteins bind peptides of eight to ten amino acids to present them at the cell surface to cytotoxic T cells. The class I binding groove binds the peptide via hydrogen bonds with the peptide termini and via diverse interactions with the anchor residue side chains of the peptide. To elucidate which of these interactions is most important for the thermodynamic and kinetic stability of the peptide-bound state, we have combined molecular dynamics simulations and experimental approaches in an investigation of the conformational dynamics and binding parameters of a murine class I molecule (H-2Kb with optimal and truncated natural peptide epitopes. We show that the F pocket region dominates the conformational and thermodynamic properties of the binding groove, and that therefore the binding of the C terminus of the peptide to the F pocket region plays a crucial role in bringing about the peptide-bound state of MHC class I.

  3. Evidence for auto-inhibition by the N terminus of hADAR2 and activation by dsRNA binding. (United States)

    Macbeth, Mark R; Lingam, Arunth T; Bass, Brenda L


    Adenosine deaminases that act on RNA (ADARs) catalyze adenosine to inosine conversion in RNA that is largely double stranded. Human ADAR2 (hADAR2) contains two double-stranded RNA binding motifs (dsRBMs), separated by a 90-amino acid linker, and these are followed by the C-terminal catalytic domain. We assayed enzymatic activity of N-terminal deletion constructs of hADAR2 to determine the role of the dsRBMs and the intervening linker peptide. We found that a truncated protein consisting of one dsRBM and the deaminase domain was capable of deaminating a short 15-bp substrate. In contrast, full-length hADAR2 was inactive on this short substrate. In addition, we observed that the N terminus, which was deleted from the truncated protein, inhibits editing activity when added in trans. We propose that the N-terminal domain of hADAR2 contains sequences that cause auto-inhibition of the enzyme. Our results suggest activation requires binding to an RNA substrate long enough to accommodate interactions with both dsRBMs. Copyright 2004 RNA Society

  4. Synthesis and properties of peptide nucleic acid labeled at the N-terminus with HiLyte Fluor 488 fluorescent dye. (United States)

    Hnedzko, Dziyana; McGee, Dennis W; Rozners, Eriks


    Fluorescently labeled peptide nucleic acids (PNAs) are important tools in fundamental research and biomedical applications. However, synthesis of labeled PNAs, especially using modern and expensive dyes, is less explored than similar preparations of oligonucleotide dye conjugates. Herein, we present a simple procedure for labeling of the PNA N-terminus with HiLyte Fluor 488 as the last step of solid phase PNA synthesis. A minimum excess of 1.25equiv of activated carboxylic acid achieved labeling yields close to 90% providing a good compromise between the price of dye and the yield of product and significant improvement over previous literature procedures. The HiLyte Fluor 488-labeled PNAs retained the RNA binding ability and in live cell fluorescence microscopy experiments were brighter and significantly more photostable than PNA labeled with carboxyfluorescein. In contrast to fluorescein-labeled PNA, the fluorescence of PNAs labeled with HiLyte Fluor 488 was independent of pH in the biologically relevant range of 5-8. The potential of HiLyte Fluor 488-labeling for studies of PNA cellular uptake and distribution was demonstrated in several cell lines. Copyright © 2016 Elsevier Ltd. All rights reserved.

  5. [Create or copy... Which is the difference?]. (United States)

    López P, Ricardo


    Creating and copying are two different processes; we must not confuse creativity with plagiarism. However, this distinction is problematic, because there is no possibility of creating from scratch, this implies that any creative act necessarily arises from accumulative experience, inevitably producing a continuity between old and new. Even so it is necessary to establish clearly the difference between creating and copying. It is not desirable that a matter of such importance remains in the nebula or that the relationship between creativity and ethics is kept unaware. There are many cases of plagiarism, but this cannot be a consolation. There is no gain when the existence of a plagiarism is ignored or concealed and less when it is unjustified.

  6. Can Physical Examination Create a Stener Lesion? (United States)

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


    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.

  7. How to create a serious game?

    Directory of Open Access Journals (Sweden)

    O. Heidmann


    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.

  8. Creating e-learning games with Unity

    CERN Document Server

    Horachek, David


    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.

  9. Creating dynamic UI with Android fragments

    CERN Document Server

    Wilson, Jim


    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.

  10. Creating a Website The Missing Manual

    CERN Document Server

    MacDonald, Matthew


    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

  11. Creating a web site the missing manual

    CERN Document Server

    MacDonald, Matthew


    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

  12. Creating Gaze Annotations in Head Mounted Displays

    DEFF Research Database (Denmark)

    Mardanbeigi, Diako; Qvarfordt, Pernilla


    , the user simply captures an image using the HMD’s camera, looks at an object of interest in the image, and speaks out the information to be associated with the object. The gaze location is recorded and visualized with a marker. The voice is transcribed using speech recognition. Gaze annotations can......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...

  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 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 ...

  14. Creating sustainable environmental management in Senegal's cities ...

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


    Apr 28, 2016 ... IAGU specializes in action research, technical support, and information on the urban environment including urban agriculture, solid waste management, strategic environmental planning, and urban risk management. It works with African city administrations to create sustainable, participatory systems for ...

  15. Mental Mapping: A Lesson that Creates Itself (United States)

    Comenetz, Joshua


    Mental image and place-preference maps of college students in Florida were created through a two-part lesson. The patterns revealed by these maps were linked to students' life experiences, census data on migration and income, and similar studies conducted in other states. Students prefer states with established migration links to Florida and…

  16. Creating a Positive Classroom Environment for Children. (United States)

    Prutzman, Priscilla


    Describes cooperation and affirmation activities designed to create a classroom environment in which children interact peacefully and resolve conflicts creatively. Exercises include cooperation drawing, group hand prints and shadow murals, a circle game entitled "Rainstorm," student interviews, affirmation shows using video, affirmation clapping,…

  17. Creating Career Awareness Among Secondary School Students ...

    African Journals Online (AJOL)

    This paper discussed the need for creating career awareness among Secondary School Students through career fair Career selection is a critical issue for secondary school students who may not be aware of the existing occupations in the labour market. It is important that career information be made available to students ...

  18. Understanding Critical Thinking to Create Better Doctors (United States)

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


    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…

  19. Creating a Character as a Writing Exercise. (United States)

    Latta, Alan D.


    Suggests an approach to teaching German as a second language in which students jointly create a character and her family and then write a series of stories during he year about the character in various everyday situations. Benefits include use of basic language, cultural issues, and stimulation of student creativity and interest. (Author/VWL)

  20. Creating and decomposing vector Bessel beams

    CSIR Research Space (South Africa)

    Dudley, Angela L


    Full Text Available -1 58th Annual SAIP Conference, Richards Bay, 8-12 July 2013 Creating and decomposing vector Bessel beams Angela Dudley1,*, Yanming Li2, Thandeka Mhlanga1, Michael Escuti2 and Andrew Forbes1 1 CSIR National Laser Centre, Pretoria, South...

  1. Creating, Invigorating, and Sustaining Effective Teams. (United States)

    Trimble, Susan; Miller, John W.


    Teams can boost creativity, morale, and communication, but they can also unleash disharmony, create tension, and waste time. To maximize teaming benefits, administrators must share authority, cultivate teacher leadership, train all team members, use situational leadership, model effective team leader behaviors, provide incentives, support each…

  2. Creating Music Environments in Early Childhood Programs. (United States)

    Achilles, Elayne


    Describes how teachers and caregivers can create music environments in early childhood settings that connect to other areas of development. Discusses how music environments can accommodate free-choice participation, describes the caregiver's role, and suggests music activities. Includes definitions of musical concepts for young children, also tips…

  3. The Media Creates Us in Its Image (United States)

    Stivers, Richard


    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.

  4. Extending the "Knowledge Advantage": Creating Learning Chains (United States)

    Maqsood, Tayyab; Walker, Derek; Finegan, Andrew


    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…

  5. Creating Future Memories: A Dialogue on Process

    DEFF Research Database (Denmark)

    Rehder, Mads Middelboe


    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 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 ... Country(s). Middle East, North of Sahara, South of Sahara, Central Asia, Far East Asia, South Asia, Palestine ...

  7. Creating a Data Warehouse using SQL Server

    DEFF Research Database (Denmark)

    Sørensen, Jens Otto; Alnor, Karl


    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...

  8. Introduction: Leadership and Diversity: Creating Inclusive Schools. (United States)

    Madsen, Jean A.; Mabokela, Reitumetse Obakeng


    Introduces a theme issue that explores the perceptions of teachers toward African American school leaders as they seek to create inclusive school environments; the capacity of teachers and school leaders to negotiate intergroup conflict so they can work collaboratively on matters of race; and leaders' interpretation and implementation of policies…

  9. Creating social impact with sport events

    NARCIS (Netherlands)

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


    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

  10. Humanising education through technology: creating social presence ...

    African Journals Online (AJOL)

    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 control. This article argues the importance of social presence as a building block of successful learning environments ...

  11. Creating Teams Increases Extension Educator Productivity (United States)

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


    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…

  12. Green Energy Technologies Create Green Jobs

    Energy Technology Data Exchange (ETDEWEB)



    The U.S. Department of Energy (DOE) is developing advanced energy technologies that can help address climate change and reduce U.S. dependence on oil. As these new technologies are launched into commercial use, they create new jobs for American workers.

  13. Engineering Encounters: Creating a Prosthetic Hand (United States)

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


    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…

  14. Creating Learning Communities in the Classroom (United States)

    Saville, Bryan K.; Lawrence, Natalie Kerr; Jakobsen, Krisztina V.


    There are many ways to construct classroom-based learning communities. Nevertheless, the emphasis is always on cooperative learning. In this article, the authors focus on three teaching methods--interteaching, team-based learning, and cooperative learning in large, lecture-based courses--that they have used successfully to create classroom-based…

  15. Creating seamless connections: Intersecting the social and ...

    African Journals Online (AJOL)

    university's effort to create intimate learning communities for students within residence halls as a way of integrating students' academic experiences with their residential lives. Both examples illustrate how learning becomes seamless and continues beyond the classroom by allowing students to be organised into learning ...

  16. Creating Smart-er Cities: An Overview (United States)

    Allwinkle, Sam; Cruickshank, Peter


    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. Photocatalytic Solutions Create Self-Cleaning Surfaces (United States)


    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.

  18. Creating sustainable empowering learning environments through ...

    African Journals Online (AJOL)

    Creating sustainable empowering learning environments through scholarship of engagement. ... South African Journal of Higher Education ... Abstract. The assumption grounding this issue of SAJHE is that; a university or any institution of higher learning comes to its fullness through serious engagement with the community.

  19. Creating Self-Portraits. ERIC Digest. (United States)

    Redekopp, Dave E.; And Others

    Creating Self-Portraits is an individual and/or group career development tool designed to assess without testing. Researchers have found that testing can be counter-productive; once clients were labelled, they frequently stopped self-examination. A tool was needed that would help people understand themselves in a way that would encourage further…

  20. Creating the networking enterprises - logistics determinants

    Directory of Open Access Journals (Sweden)

    Ewa Kulińska


    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.

  1. Creating by Reusing Learning Design Solutions

    NARCIS (Netherlands)

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


    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

  2. Creating Brand India: Strategies, Issues, and Challenges

    National Research Council Canada - National Science Library

    Alok Chakrawal; Pratibha Goyal


    .... Bureaucracy, corruption, delay in clearance of business proposals, ethical standards and work culture, tax reforms, political interventions, socio-economic barriers, regionalism, etc. are some of the challenges that must be coped with. This article examines these various issues and challenges that must be overcome in creating Brand India.

  3. Creating Appropriate Graphics for Business Situations (United States)

    Katz, Susan M.


    Charts and graphs are ubiquitous in business documents, and most students in the author's business communication courses are well aware that they need to be able to create many different types of data representation. Most of them have had a great deal of experience working with spreadsheet applications, and they know how to manipulate data and…

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

    African Journals Online (AJOL)


    May 1, 2017 ... Using the Sustainable Livelihoods Concept and a qualitative approach, this study sought to understand ... Towards Creating Sustainable Ecotourism Interventions: Practical Lessons from Mesomagoro, Ghana. Africa, was .... as mortars, pestles, stools, drums chewing sticks, sponges and material for houses;.

  5. Crystal structure of the Habc domain of neuronal syntaxin from the squid Loligo pealei reveals conformational plasticity at its C-terminus

    Directory of Open Access Journals (Sweden)

    Bracher Andreas


    Full Text Available Abstract Background Intracellular membrane fusion processes are mediated by the spatial and temporal control of SNARE complex assembly that results in the formation of a four-helical bundle, composed of one vesicle SNARE and three target membrane SNARE polypeptide chains. Syntaxins are essential t-SNAREs and are characterized by an N-terminal Habc domain, a flexible linker region, a coiled-coil or SNARE motif and a membrane anchor. The N-terminal Habc domain fulfills important regulatory functions while the coiled-coil motif, present in all SNAREs, is sufficient for SNARE complex formation, which is thought to drive membrane fusion. Results Here we report the crystal structure of the Habc domain of neuronal syntaxin from the squid Loligo pealei, s-syntaxin. Squid Habc crystallizes as a dimer and the monomer structure consists of a three-helical bundle. One molecule is strikingly similar to mammalian syntaxin 1A while the second one shows a structural deviation from the common fold in that the C-terminal part of helix C unwinds and adopts an extended conformation. Conclusion Conservation of surface residues indicates that the cytosolic part of s-syntaxin can adopt an auto-inhibitory closed conformation that may bind squid neuronal Sec1, s-Sec1, in the same manner as observed in structure of the rat nSec1/syntaxin 1A complex. Furthermore, despite the overall structural similarity, the observed changes at the C-terminus of one molecule indicate structural plasticity in neuronal syntaxin. Implications of the structural conservation and the changes are discussed with respect to potential Habc domain binding partners such as Munc13, which facilitates the transition from the closed to the open conformation.

  6. Distinct roles of Ser-764 and Lys-773 at the N terminus of von Willebrand factor in complex assembly with coagulation factor VIII. (United States)

    Castro-Núñez, Lydia; Bloem, Esther; Boon-Spijker, Mariëtte G; van der Zwaan, Carmen; van den Biggelaar, Maartje; Mertens, Koen; Meijer, Alexander B


    Complex formation between coagulation factor VIII (FVIII) and von Willebrand factor (VWF) is of critical importance to protect FVIII from rapid in vivo clearance and degradation. We have now employed a chemical footprinting approach to identify regions on VWF involved in FVIII binding. To this end, lysine amino acid residues of VWF were chemically modified in the presence of FVIII or activated FVIII, which does not bind VWF. Nano-LC-MS analysis showed that the lysine residues of almost all identified VWF peptides were not differentially modified upon incubation of VWF with FVIII or activated FVIII. However, Lys-773 of peptide Ser-766-Leu-774 was protected from chemical modification in the presence of FVIII. In addition, peptide Ser-764-Arg-782, which comprises the first 19 amino acid residues of mature VWF, showed a differential modification of both Lys-773 and the α-amino group of Ser-764. To verify the role of Lys-773 and the N-terminal Ser-764 in FVIII binding, we employed VWF variants in which either Lys-773 or Ser-764 was replaced with Ala. Surface plasmon resonance analysis and competition studies revealed that VWF(K773A) exhibited reduced binding to FVIII and the FVIII light chain, which harbors the VWF-binding site. In contrast, VWF(S764A) revealed more effective binding to FVIII and the FVIII light chain compared with WT VWF. The results of our study show that the N terminus of VWF is critical for the interaction with FVIII and that Ser-764 and Lys-773 have opposite roles in the binding mechanism.

  7. Inhibition of Macrophage Functions by the C-Terminus of Murine S100A9 Is Dependent on B-1 Cells

    Directory of Open Access Journals (Sweden)

    Rosana Lima Pagano


    Full Text Available The protein S100A9 plays a key role in the control of inflammatory response. The C-terminus of the murine S100A9 protein (mS100A9p downregulates the spreading and phagocytic activity of adherent peritoneal cells. Murine peritoneal cells are constituted by macrophages and B-1 cells, and the latter exert an inhibitory effect on macrophage functions by secreting interleukin- (IL- 10. Here, we investigated the influence of B-1 cells on the inhibitory effect evoked by mS100A9p on macrophages. mS100A9p did not alter spreading and phagocytosis either by peritoneal macrophages obtained from mice deprived of B-1 cells or by bone marrow-derived macrophages (BMDMϕ. Nevertheless, when BMDMϕ were cocultivated by direct or indirect contact with B-1 cells treated with mS100A9p, the phagocytosis by BMDMϕ was decreased, showing that the effect of mS100A9p on macrophages was modulated by B-1 cells and/or their secretory compounds. Furthermore, the inhibitory action of mS100A9p on phagocytosis by adherent peritoneal cells was abolished in cells obtained from IL-10 knockout mice. Taken together, the results show that mS100A9p has no direct inhibitory effect on macrophages; however, mS100A9p modulates B-1 cells, which in turn downregulates macrophages, at least in part, via IL-10. These data contribute to the characterization of S100A9 functions involving B-1 cells in the regulation of the inflammatory process.

  8. 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.

  9. Engineering the expression and characterization of two novel laccase isoenzymes from Coprinus comatus in Pichia pastoris by fusing an additional ten amino acids tag at N-terminus. (United States)

    Gu, Chunjuan; Zheng, Fei; Long, Liangkun; Wang, Jing; Ding, Shaojun


    The detail understanding of physiological/biochemical characteristics of individual laccase isoenzymes in fungi is necessary for fundamental and application purposes, but our knowledge is still limited for most of fungi due to difficult to express laccases heterologously. In this study, two novel laccase genes, named lac3 and lac4, encoding proteins of 547 and 532-amino acids preceded by 28 and 16-residue signal peptides, respectively, were cloned from the edible basidiomycete Coprinus comatus. They showed 70% identity but much lower homology with other fungal laccases at protein level (less than 58%). Two novel laccase isoenzymes were successfully expressed in Pichia pastoris by fusing an additional 10 amino acids (Thr-Pro-Phe-Pro-Pro-Phe-Asn-Thr-Asn-Ser) tag at N-terminus, and the volumetric activities could be dramatically enhanced from undetectable level to 689 and 1465 IU/l for Lac3 and Lac4, respectively. Both laccases possessed the lowest Km and highest kcat/Km value towards syringaldazine, followed by ABTS, guaiacol and 2,6-dimethylphenol similar as the low redox potential laccases from other microorganisms. Lac3 and Lac4 showed resistant to SDS, and retained 31.86% and 43.08% activity in the presence of 100 mM SDS, respectively. Lac3 exhibited higher decolorization efficiency than Lac4 for eleven out of thirteen different dyes, which may attribute to the relatively higher catalytic efficiency of Lac3 than Lac4 (in terms of kcat/Km) towards syringaldazine and ABTS. The mild synergistic decolorization by two laccases was observed for triphenylmethane dyes but not for anthraquinone and azo dyes.

  10. Engineering the expression and characterization of two novel laccase isoenzymes from Coprinus comatus in Pichia pastoris by fusing an additional ten amino acids tag at N-terminus.

    Directory of Open Access Journals (Sweden)

    Chunjuan Gu

    Full Text Available The detail understanding of physiological/biochemical characteristics of individual laccase isoenzymes in fungi is necessary for fundamental and application purposes, but our knowledge is still limited for most of fungi due to difficult to express laccases heterologously. In this study, two novel laccase genes, named lac3 and lac4, encoding proteins of 547 and 532-amino acids preceded by 28 and 16-residue signal peptides, respectively, were cloned from the edible basidiomycete Coprinus comatus. They showed 70% identity but much lower homology with other fungal laccases at protein level (less than 58%. Two novel laccase isoenzymes were successfully expressed in Pichia pastoris by fusing an additional 10 amino acids (Thr-Pro-Phe-Pro-Pro-Phe-Asn-Thr-Asn-Ser tag at N-terminus, and the volumetric activities could be dramatically enhanced from undetectable level to 689 and 1465 IU/l for Lac3 and Lac4, respectively. Both laccases possessed the lowest Km and highest kcat/Km value towards syringaldazine, followed by ABTS, guaiacol and 2,6-dimethylphenol similar as the low redox potential laccases from other microorganisms. Lac3 and Lac4 showed resistant to SDS, and retained 31.86% and 43.08% activity in the presence of 100 mM SDS, respectively. Lac3 exhibited higher decolorization efficiency than Lac4 for eleven out of thirteen different dyes, which may attribute to the relatively higher catalytic efficiency of Lac3 than Lac4 (in terms of kcat/Km towards syringaldazine and ABTS. The mild synergistic decolorization by two laccases was observed for triphenylmethane dyes but not for anthraquinone and azo dyes.

  11. Comparative evaluation of tumor targeting using the anti-HER2 ADAPT scaffold protein labeled at the C-terminus with indium-111 or technetium-99m. (United States)

    Garousi, Javad; Lindbo, Sarah; Mitran, Bogdan; Buijs, Jos; Vorobyeva, Anzhelika; Orlova, Anna; Tolmachev, Vladimir; Hober, Sophia


    ABD-Derived Affinity Proteins (ADAPTs) is a novel class of engineered scaffold proteins derived from an albumin-binding domain of protein G. The use of ADAPT6 derivatives as targeting moiety have provided excellent preclinical radionuclide imaging of human epidermal growth factor 2 (HER2) tumor xenografts. Previous studies have demonstrated that selection of nuclide and chelator for its conjugation has an appreciable effect on imaging properties of scaffold proteins. In this study we performed a comparative evaluation of the anti-HER2 ADAPT having an aspartate-glutamate-alanine-valine-aspartate-alanine-asparagine-serine (DEAVDANS) N-terminal sequence and labeled at C-terminus with (99m)Tc using a cysteine-containing peptide based chelator, glycine-serine-serine-cysteine (GSSC), and a similar variant labeled with (111)In using a maleimido derivative of 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA) chelator. Both (99m)Tc-DEAVDANS-ADAPT6-GSSC and (111)In-DEAVDANS-ADAPT6-GSSC-DOTA accumulated specifically in HER2-expressing SKOV3 xenografts. The tumor uptake of both variants did not differ significantly and average values were in the range of 19-21%ID/g. However, there was an appreciable variation in uptake of conjugates in normal tissues that resulted in a notable difference in the tumor-to-organ ratios. The (111)In-DOTA label provided 2-6 fold higher tumor-to-organ ratios than (99m)Tc-GSSC and is therefore the preferable label for ADAPTs.

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

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


    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 (SdrEN2N3) with high affinity, and determined the crystal structures of apo-SdrEN2N3 and the SdrEN2N3-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, 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. © 2017 The Author(s).


    Directory of Open Access Journals (Sweden)

    Kristijan Posavec


    Full Text Available Paper presents automated technique for creating lithologic log plots. Technique is based on three computer tools: Microsoft (MS Access program, LogPlot program, and Visual Basic (VB macros for MS Excel. MS Access ensures professional storage of lithologic data which can be in that way easier and faster entered, searched, updated, and also used for different purposes, while LogPlot provides tools for creating lithologic log plots. VB macros enable transfer of lithologic data from MS Access to LogPlot. Data stored in MS Access are exported in ASCII files which are later used by LogPlot for creation of lithologic log plots. Presented concept facilitates creation of lithologic log plots, and automated technique enables processing of a large number of data i.e. creation of lareg number lithologic log plots in a short period of time (the paper is published in Croatian.

  14. Creating a culture for information systems success

    CERN Document Server

    Belkhamza, Zakariya


    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...

  15. Creating and Editing Video to Accompany Manuscripts. (United States)

    Gordon, Shayna L; Porto, Dennis A; Ozog, David M; Council, M Laurin


    The use of video can enhance the learning experience by demonstrating procedural techniques that are difficult to relay in writing. Several peer-reviewed journals allow publication of videos alongside articles to complement the written text. The purpose of this article is to instruct the dermatologic surgeon on how to create and edit a video using a smartphone, to accompany a article. The authors describe simple tips to optimize surgical videography. The video that accompanies this article further demonstrates the techniques described. Creating a surgical video requires little experience or equipment and can be completed in a modest amount of time. Making and editing a video to accompany a article can be accomplished by following the simple recommendations in this article. In addition, the increased use of video in dermatologic surgery education can enhance the learning opportunity.

  16. Novel device for creating continuous curvilinear capsulorhexis. (United States)

    Soylak, Mustafa


    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.

  17. How do entrepreneurs think they create value?

    DEFF Research Database (Denmark)

    Lyth Frederiksen, Dennis; Brem, Alexander


    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 entrepren......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...... entrepreneurs have read the book and may have followed his advice. Hence, we investigate the merits and characteristics of the methods detailed by Ries through a comparison with leading theories and empirical evidence found in the scientific literature. The results indicate that overall the methods find...

  18. Designing value-creating supply chain networks

    CERN Document Server

    Martel, Alain


    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...

  19. Creating interdisciplinary education within monodisciplinary structures

    DEFF Research Database (Denmark)

    Lindvig, Katrine; Lyall, Catherine; R. Meagher, Laura


    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......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....... 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...

  20. Barriers to creating a secure MPI

    Energy Technology Data Exchange (ETDEWEB)

    Brightwell, R.; Greenberg, D.S.; Matt, B.J. [Sandia National Labs., Albuquerque, NM (United States); Davida, G.I. [Univ. of Wisconsin, Milwaukee, WI (United States). Dept. of Computer Sciences


    This paper explores some of the many issues in developing security enhanced MPI for embedded real-time systems supporting the Department of Defense`s Multi-level Security policy (DoD MLS) are presented along with the preliminary design for such an MPI variant. In addition some of the many issues that need to be addressed in creating security enhanced versions of MPI for other domains are discussed. 19 refs.

  1. Creating an Asthma-Friendly School

    Centers for Disease Control (CDC) Podcasts


    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.

  2. Climate Change Creates Trade Opportunity in India


    Dinda, Soumyananda


    Climate change is an emerging challenge to developing economy like India however it also creates opportunity to grow through climate friendly goods production and new direction of trade. This paper focuses India’s potential export trade in climate friendly goods. The estimated gravity model is defined as the potential trade and potential trade gap is measured as how well a bilateral trade flow performs relative to the mean as predicted by the model. Potential trade gap means that actual trade...

  3. Cognitive abilities and creating metaphorical names

    Directory of Open Access Journals (Sweden)

    Avanesyan, Marina O.


    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.

  4. How to create a UX story


    Michailidou, Ioanna; von Saucken, Constantin; Kremer, Simon; Lindemann, Udo


    Narratives are a tool used in many disciplines. In the area of User Experience Design (UXD), in particular, a storytelling approach can be applied during the whole design process to improve the quality of developed concepts regarding user experience (UX). Furthermore stories support designers in ex-ploring and communicating their new concept ideas. However, the guidelines on how to create a story are either too abstract or do not focus on the experience elements of the interaction. This paper...

  5. Creating an international curriculum: why and how?


    Magne, PJ


    Today’s globally interconnected world offers a vast array of new opportunities, but it has simultaneously created a need for greater intercultural understanding (Koehne, 2006). The current rise of internationalisation policy and strategy suggests that HEIs recognise the part they need to play to enable graduates to operate effectively in the 21st century (Bremer & Van-der-Wende, 1995; Knight & Yorke, 2003; Shiel, 2006) Whilst it is would be irresponsible to ignore the challenges posed by inte...

  6. Can cognitive science create a cognitive economics? (United States)

    Chater, Nick


    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.

  7. Creating an Online Training Module on RDM


    Guy, Marieke; Cope, Jez; Pink, Catherine


    Creating an Online Training Module on Research Data Management for the University of BathResearch 360In 2011 the University of Bath was awarded funding by the JISC Managing Research Data Programme to support and develop Research Data Management across the institution.The Research 360: Managing data across the institutional research lifecycle project (Research360@Bath) will develop policies, infrastructure and training resources to help researchers at the University of Bath to get the most out...

  8. Creating social impact with sport events


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


    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 excitement, pleasure and feelings of camaraderie are experienced among millions of people in the case of mega events. Still, a fairly large section of the population does not care that much for sports. Som...

  9. Creating an Innovative Attitude at Work

    DEFF Research Database (Denmark)

    Eriksen, Kaare; Tollestrup, Christian


    or tacit knowledge among colleagues in a complex organization. The set-up of the project was organized as a series of workshops also involving design students, and the paper outlines the difficulties and results from the initiative. The project showed that designerly methods can be very effective...... in creating the participants’ positive innovative approach, but also that it is a challenge to translate such methods and vocabulary, and more trained designerly assistance might need to be applied in the process....

  10. Public libraries: places creating social capital?


    Vårheim, Andreas


    Purpose – The purpose of this paper is to show why studies of public libraries, regarding their possible contribution in creating social capital, are important for social capital research in general, and are important for library practice in particular. Design/methodology/approach – Building on the latest theoretical developments and empirical findings of social capital research, the role of the public library as a potential creator of social capital is discussed. Findings from both quantita...

  11. Creating marketing strategies for higher education institutions


    Lidia Białoń


    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...

  12. Evolution of the Twist Subfamily Vertebrate Proteins: Discovery of a Signature Motif and Origin of the Twist1 Glycine-Rich Motifs in the Amino-Terminus Disordered Domain.

    Directory of Open Access Journals (Sweden)

    Yacidzohara Rodriguez

    Full Text Available Twist proteins belong to the basic helix-loop-helix (bHLH family of multifunctional transcriptional factors. These factors are known to use domains other than the common bHLH in protein-protein interactions. There has been much work characterizing the bHLH domain and the C-terminus in protein-protein interactions but despite a few attempts more focus is needed at the N-terminus. Since the region of highest diversity in Twist proteins is the N-terminus, we analyzed the conservation of this region in different vertebrate Twist proteins and study the sequence differences between Twist1 and Twist2 with emphasis on the glycine-rich regions found in Twist1. We found a highly conserved sequence motif in all Twist1 (SSSPVSPADDSLSNSEEE and Twist2 (SSSPVSPVDSLGTSEEE mammalian species with unknown function. Through sequence comparison we demonstrate that the Twist protein family ancestor was "Twist2-like" and the two glycine-rich regions found in Twist1 sequences were acquired late in evolution, apparently not at the same time. The second glycine-rich region started developing first in the fish vertebrate group, while the first glycine region arose afterwards within the reptiles. Disordered domain and secondary structure predictions showed that the amino acid sequence and disorder feature found at the N-terminus is highly evolutionary conserved and could be a functional site that interacts with other proteins. Detailed examination of the glycine-rich regions in the N-terminus of Twist1 demonstrate that the first region is completely aliphatic while the second region contains some polar residues that could be subject to post-translational modification. Phylogenetic and sequence space analysis showed that the Twist1 subfamily is the result of a gene duplication during Twist2 vertebrate fish evolution, and has undergone more evolutionary drift than Twist2. We identified a new signature motif that is characteristic of each Twist paralog and identified

  13. Evolution of the Twist Subfamily Vertebrate Proteins: Discovery of a Signature Motif and Origin of the Twist1 Glycine-Rich Motifs in the Amino-Terminus Disordered Domain. (United States)

    Rodriguez, Yacidzohara; Gonzalez-Mendez, Ricardo R; Cadilla, Carmen L


    Twist proteins belong to the basic helix-loop-helix (bHLH) family of multifunctional transcriptional factors. These factors are known to use domains other than the common bHLH in protein-protein interactions. There has been much work characterizing the bHLH domain and the C-terminus in protein-protein interactions but despite a few attempts more focus is needed at the N-terminus. Since the region of highest diversity in Twist proteins is the N-terminus, we analyzed the conservation of this region in different vertebrate Twist proteins and study the sequence differences between Twist1 and Twist2 with emphasis on the glycine-rich regions found in Twist1. We found a highly conserved sequence motif in all Twist1 (SSSPVSPADDSLSNSEEE) and Twist2 (SSSPVSPVDSLGTSEEE) mammalian species with unknown function. Through sequence comparison we demonstrate that the Twist protein family ancestor was "Twist2-like" and the two glycine-rich regions found in Twist1 sequences were acquired late in evolution, apparently not at the same time. The second glycine-rich region started developing first in the fish vertebrate group, while the first glycine region arose afterwards within the reptiles. Disordered domain and secondary structure predictions showed that the amino acid sequence and disorder feature found at the N-terminus is highly evolutionary conserved and could be a functional site that interacts with other proteins. Detailed examination of the glycine-rich regions in the N-terminus of Twist1 demonstrate that the first region is completely aliphatic while the second region contains some polar residues that could be subject to post-translational modification. Phylogenetic and sequence space analysis showed that the Twist1 subfamily is the result of a gene duplication during Twist2 vertebrate fish evolution, and has undergone more evolutionary drift than Twist2. We identified a new signature motif that is characteristic of each Twist paralog and identified important residues within

  14. 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)


    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)

  15. 10 ways to create shareholder value. (United States)

    Rappaport, Alfred


    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.

  16. How leaders create and use networks. (United States)

    Ibarra, Herman; Hunter, Mark


    Most people acknowledge that networking-creating a fabric of personal contacts to provide support, feedback, insight, and resources--is an essential activity for an ambitious manager. Indeed, it's a requirement even for those focused simply on doing their current jobs well. For some, this is a distasteful reality. Working through networks, they believe,means relying on "who you know" rather than "what you know"--a hypocritical, possibly unethical, way to get things done. But even people who understand that networking is a legitimate and necessary part of their jobs can be discouraged by the payoff--because they are doing it in too limited a fashion. On the basis of a close study of 30 emerging leaders, the authors outline three distinct forms of networking. Operational networking is geared toward doing one's assigned tasks more effectively. It involves cultivating stronger relationships with colleagues whose membership in the network is clear; their roles define them as stakeholders. Personal networking engages kindred spirits from outside an organization in an individual's efforts to learn and find opportunities for personal advancement. Strategic networking puts the tools of networking in the service of business goals. At this level, a manager creates the kind of network that will help uncover and capitalize on new opportunities for the company. The ability to move to this level of networking turns out to be a key test of leadership. Companies often recognize that networks are valuable, andthey create explicit programs to support them. But typically these programs facilitate only operational networking. Likewise, industry associations provide formal contexts for personal networking. The unfortunate effect is to give managers the impression that they know how to network and are doing so sufficiently. A sidebar notes the implication for companies' leadership development initiatives: that teaching strategic networking skills will serve their aspiring leaders and

  17. Workshop: Creating Your Institutional Research Repository

    KAUST Repository

    Grenz, Daryl M.


    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.

  18. Creating environments that foster academic integrity. (United States)

    Tippitt, Michelle Pixley; Ard, Nell; Kline, Juanita Reese; Tilghman, Joan; Chamberlain, Barbara; Meagher, P Gail


    A number of studies related to academic dishonesty within the nursing student population have been published; however, little has been written in the nursing literature regarding academic integrity and means of promoting this value. In addition to the many short-term solutions to prevent cheating and dissuade academic misconduct that are offered, solutions that promote long-term affective changes underlying the acquisition of academic integrity are needed. This article provides a context for discussions related to academic integrity, explores issues facing faculty when dealing with this challenge, and offers short-term and long-term strategies for creating environments that foster academic integrity.

  19. Creating a Library Database Search using Drupal

    Directory of Open Access Journals (Sweden)

    Danielle M. Rosenthal


    Full Text Available When Florida Gulf Coast University Library was faced with having to replace its database locator, they needed to find a low-cost, non-staff intensive replacement for their 350 plus databases search tool. This article details the development of a library database locator, based on the methods described in Leo Klein’s “Creating a Library Database Page using Drupal” online presentation. The article describes how the library used Drupal along with several modules, such as CCK, Views, and FCKeditor. It also discusses various Drupal search modules that were evaluated during the process.

  20. Creating an organizational climate for multiculturalism. (United States)

    Bruhn, J G


    Multiculturism is an ideal goal for our society, its organizations, and its institutions, involving a continuous process of education and change within organizations. Multiculturalism begins with diversity and requires various steps to achieve changes in attitudes, behaviors, and values. The leadership of organizations must not only commit to diversification, but they must participate in it and reward its efforts. Diversification should be managed by creating a climate of open participation, feedback, and control at the lower organizational levels. To micromanage the process of becoming diverse increases resistance and paranoia and counters educational efforts.

  1. Does the New Economy Create Higher Productivity?

    DEFF Research Database (Denmark)

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


    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...... IT use, has brought focus on the IT industry as a catalyst for growth. The Danish Ministry of Finance (2001) points out general increased productivity in Denmark at macro level and this increase is said to be a result of increased IT use. The question is, however, if the influence of IT investments...

  2. 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....

  3. Green electronics manufacturing creating environmental sensible products

    CERN Document Server

    Wang, John X


    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

  4. Creating Posters for Effective Scientific Communication. (United States)

    Bavdekar, Sandeep B; Vyas, Shruti; Anand, Varun


    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.

  5. Ubiquitinylation of α-Synuclein by Carboxyl Terminus Hsp70-Interacting Protein (CHIP) Is Regulated by Bcl-2-Associated Athanogene 5 (BAG5) (United States)

    Chau, Hien; Lozano, Andres M.; Hyman, Bradley T.; McLean, Pamela J.


    Parkinson's disease (PD) is a common neurodegenerative condition in which abnormalities in protein homeostasis, or proteostasis, may lead to accumulation of the protein α-synuclein (α-syn). Mutations within or multiplications of the gene encoding α-syn are known to cause genetic forms of PD and polymorphisms in the gene are recently established risk factors for idiopathic PD. α-syn is a major component of Lewy bodies, the intracellular proteinaceous inclusions which are pathological hallmarks of most forms of PD. Recent evidence demonstrates that α-syn can self associate into soluble oligomeric species and implicates these α-syn oligomers in cell death. We have previously shown that carboxyl terminus of Hsp70-interacting protein (CHIP), a co-chaperone molecule with E3 ubiquitin ligase activity, may reduce the levels of toxic α-syn oligomers. Here we demonstrate that α-syn is ubiquitinylated by CHIP both in vitro and in cells. We find that the products from ubiquitinylation by CHIP include both monoubiquitinylated and polyubiquitinylated forms of α-syn. We also demonstrate that CHIP and α-syn exist within a protein complex with the co-chaperone bcl-2-associated athanogene 5 (BAG5) in brain. The interaction of CHIP with BAG5 is mediated by Hsp70 which binds to the tetratricopeptide repeat domain of CHIP and the BAG domains of BAG5. The Hsp70-mediated association of BAG5 with CHIP results in inhibition of CHIP E3 ubiquitin ligase activity and subsequently reduces α-syn ubiquitinylation. Furthermore, we use a luciferase-based protein-fragment complementation assay of α-syn oligomerization to investigate regulation of α-syn oligomers by CHIP in living cells. We demonstrate that BAG5 mitigates the ability of CHIP to reduce α-syn oligomerization and that non-ubiquitinylated α-syn has an increased propensity for oligomerization. Thus, our results identify CHIP as an E3 ubiquitin ligase of α-syn and suggest a novel function for BAG5 as a modulator of CHIP E3

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


    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.

  7. Involvement of a Heptad Repeat in the Carboxyl Terminus of the Dihydropyridine Receptor β1a Subunit in the Mechanism of Excitation-Contraction Coupling in Skeletal Muscle (United States)

    Sheridan, David C.; Cheng, Weijun; Carbonneau, Leah; Ahern, Chris A.; Coronado, Roberto


    Chimeras consisting of the homologous skeletal dihydropyridine receptor (DHPR) β1a subunit and the heterologous cardiac/brain β2a subunit were used to determine which regions of β1a were responsible for the skeletal-type excitation-contraction (EC) coupling phenotype. Chimeras were transiently transfected in β1 knockout myotubes and then voltage-clamped with simultaneous measurement of confocal fluo-4 fluorescence. All chimeras expressed a similar density of DHPR charge movements, indicating that the membrane density of DHPR voltage sensors was not a confounding factor in these studies. The data indicates that a β1a-specific domain present in the carboxyl terminus, namely the D5 region comprising the last 47 residues (β1a 478–524), is essential for expression of skeletal-type EC coupling. Furthermore, the location of β1aD5 immediately downstream from conserved domain D4 is also critical. In contrast, chimeras in which β1aD5 was swapped by the D5 region of β2a expressed Ca2+ transients triggered by the Ca2+ current, or none at all. A hydrophobic heptad repeat is present in domain D5 of β1a (L478, V485, V492). To determine the role of this motif, residues in the heptad repeat were mutated to alanines. The triple mutant β1a(L478A/V485A/V492A) recovered weak skeletal-type EC coupling (ΔF/Fmax = 0.4 ± 0.1 vs. 2.7 ± 0.5 for wild-type β1a). However, a triple mutant with alanine substitutions at positions out of phase with the heptad repeat, β1a(S481A/L488A/S495A), was normal (ΔF/Fmax = 2.1 ± 0.4). In summary, the presence of the β1a-specific D5 domain, in its correct position after conserved domain D4, is essential for skeletal-type EC coupling. Furthermore, a heptad repeat in β1aD5 controls the EC coupling activity. The carboxyl terminal heptad repeat of β1a might be involved in protein-protein interactions with ryanodine receptor type 1 required for DHPR to ryanodine receptor type 1 signal transmission. PMID:15298900

  8. Phosphorylation of clustered serine residues in the N-terminus of BPS domain negatively regulates formation of the complex between human Grb14 and insulin receptor. (United States)

    Taira, Junichi; Kida, Yutaka; Inatomi, Kohei; Komatsu, Hideyuki; Higashimoto, Yuichiro; Sakamoto, Hiroshi


    Growth factor receptor-bound protein 14 (Grb14) is a negative regulator of insulin receptor (IR) and is involved in a negative feedback mechanism of insulin signaling. Grb14 associates with IR and inhibits its tyrosine kinase activity through the between pleckstrin homology and Src homology-2 (BPS) domain. We previously reported that the pharmacological inhibition and knockdown of glycogen synthase kinase-3 (GSK-3) facilitates the insulin-induced complex formation of human Grb14 (hGrb14) and IR, suggesting that GSK-3 suppresses hGrb14 recruitment to IR. This study further investigated a functional phosphorylation of the serine residues in hGrb14 BPS domain, identified as putative GSK-3 targets to verify an effect of GSK-3 on the hGrb14-IR complex formation. In vitro kinase assay using the motif-derived peptides showed that the serine residues located in N-terminal (Ser358, Ser362 and Ser366) and C-terminal (Ser419 and Ser423) regions of the BPS domain were phosphorylated by GSK-3. Co-immunoprecipitation and yeast two-hybrid (Y2H) experiments suggested that the negative charges genetically introduced on the Ser358, Ser362 and Ser366 suppressed the association of hGrb14 to IR. Surface plasmon resonance experiment gave Kd values of 8 nM for recombinant hGrb14 with respect to the interaction with IR β-subunit, and this affinity was lost after the replacements of the Ser358, Ser362 and Ser366 with glutamic acid residues. Y2H experiment with the BPS domain alone; however, did not show any difference owing to the same mutations. It is therefore evident that the N-terminus of the BPS domain plays an important role in the regulation of hGrb14-IR complex formation through phosphorylation, in addition to other domains. © The Authors 2017. Published by Oxford University Press on behalf of the Japanese Biochemical Society. All rights reserved.

  9. Interaction of a synthetic peptide corresponding to the N-terminus of canine distemper virus fusion protein with phospholipid vesicles: a biophysical study. (United States)

    Aranda, Francisco J; Teruel, José A; Ortiz, Antonio


    The F protein of canine distemper virus (CDV) is a classic type I glycoprotein formed by two polypeptides, F1 and F2. The N-terminal regions of the F1 polypeptides of CDV, measles virus and other paramyxoviruses present moderate to high homology, supporting the existence of a high conservation within these structures, which emphasises its role in viral-host cell membrane fusion. This N-terminal polypeptide is usually termed the fusion peptide. The fusion peptides of most viral fusion-mediating glycoproteins contain a high proportion of hydrophobic amino acids, which facilitates its insertion into target membranes during fusion. In this work we report on the interaction of a 31-residue synthetic peptide (FP31) corresponding to the N terminus of CDV F1 protein with phospholipid membranes composed of various phospholipids, as studied by means of various biophysical techniques. FTIR investigation of FP31 secondary structure in aqueous medium and in membranes resulted in a major proportion of alpha-helical structure which increased upon membrane insertion. Differential scanning calorimetry (DSC) showed that the presence of concentrations of FP31 as low as 0.1 mol%, in mixtures with L-alpha-dimyristoylphosphatidylcholine (DMPC), L-alpha-dipalmitoylphosphatidylcholine (DPPC) and L-alpha-distearoylphosphatidylcholine (DSPC), already affected the thermotropic properties of the gel to liquid-crystalline phase transition. In mixtures with the three lipids, increasing the concentration of peptide made the pretransition to disappear, and lowered and broadened the main transition. This effect was slightly stronger as the acyl chain length of the phospholipid grew larger. In the corresponding partial phase diagrams, no immiscibilities or critical points were observed. FTIR showed that incorporation of 1 mol% of peptide in DPPC shifted the antisymmetric and symmetric CH2 stretching bands to higher values, indicating the existence of an additional disordering of the acyl chain

  10. Ubiquitinylation of α-synuclein by carboxyl terminus Hsp70-interacting protein (CHIP is regulated by Bcl-2-associated athanogene 5 (BAG5.

    Directory of Open Access Journals (Sweden)

    Lorraine V Kalia


    Full Text Available Parkinson's disease (PD is a common neurodegenerative condition in which abnormalities in protein homeostasis, or proteostasis, may lead to accumulation of the protein α-synuclein (α-syn. Mutations within or multiplications of the gene encoding α-syn are known to cause genetic forms of PD and polymorphisms in the gene are recently established risk factors for idiopathic PD. α-syn is a major component of Lewy bodies, the intracellular proteinaceous inclusions which are pathological hallmarks of most forms of PD. Recent evidence demonstrates that α-syn can self associate into soluble oligomeric species and implicates these α-syn oligomers in cell death. We have previously shown that carboxyl terminus of Hsp70-interacting protein (CHIP, a co-chaperone molecule with E3 ubiquitin ligase activity, may reduce the levels of toxic α-syn oligomers. Here we demonstrate that α-syn is ubiquitinylated by CHIP both in vitro and in cells. We find that the products from ubiquitinylation by CHIP include both monoubiquitinylated and polyubiquitinylated forms of α-syn. We also demonstrate that CHIP and α-syn exist within a protein complex with the co-chaperone bcl-2-associated athanogene 5 (BAG5 in brain. The interaction of CHIP with BAG5 is mediated by Hsp70 which binds to the tetratricopeptide repeat domain of CHIP and the BAG domains of BAG5. The Hsp70-mediated association of BAG5 with CHIP results in inhibition of CHIP E3 ubiquitin ligase activity and subsequently reduces α-syn ubiquitinylation. Furthermore, we use a luciferase-based protein-fragment complementation assay of α-syn oligomerization to investigate regulation of α-syn oligomers by CHIP in living cells. We demonstrate that BAG5 mitigates the ability of CHIP to reduce α-syn oligomerization and that non-ubiquitinylated α-syn has an increased propensity for oligomerization. Thus, our results identify CHIP as an E3 ubiquitin ligase of α-syn and suggest a novel function for BAG5 as a

  11. The cooperative function of arginine residues in the Prototype Foamy Virus Gag C-terminus mediates viral and cellular RNA encapsidation. (United States)

    Hamann, Martin V; Müllers, Erik; Reh, Juliane; Stanke, Nicole; Effantin, Gregory; Weissenhorn, Winfried; Lindemann, Dirk


    One unique feature of the foamy virus (FV) capsid protein Gag is the absence of Cys-His motifs, which in orthoretroviruses are irreplaceable for multitude functions including viral RNA genome recognition and packaging. Instead, FV Gag contains glycine-arginine-rich (GR) sequences at its C-terminus. In case of prototype FV (PFV) these are historically grouped in three boxes, which have been shown to play essential functions in genome reverse transcription, virion infectivity and particle morphogenesis. Additional functions for RNA packaging and Pol encapsidation were suggested, but have not been conclusively addressed. Here we show that released wild type PFV particles, like orthoretroviruses, contain various cellular RNAs in addition to viral genome. Unlike orthoretroviruses, the content of selected cellular RNAs in capsids of PFV vector particles was not altered by viral genome encapsidation. Deletion of individual GR boxes had only minor negative effects (2 to 4-fold) on viral and cellular RNA encapsidation over a wide range of cellular Gag to viral genome ratios examined. Only the concurrent deletion of all three PFV Gag GR boxes, or the substitution of multiple arginine residues residing in the C-terminal GR box region by alanine, abolished both viral and cellular RNA encapsidation (>50 to >3,000-fold reduced), independent of the viral production system used. Consequently, those mutants also lacked detectable amounts of encapsidated Pol and were non-infectious. In contrast, particle release was reduced to a much lower extent (3 to 20-fold). Taken together, our data provides the first identification of a full-length PFV Gag mutant devoid in genome packaging and the first report of cellular RNA encapsidation into PFV particles. Our results suggest that the cooperative action of C-terminal clustered positively charged residues, present in all FV Gag proteins, is the main viral protein determinant for viral and cellular RNA encapsidation. The viral genome

  12. Creating the Medical Schools of the Future. (United States)

    Skochelak, Susan E; Stack, Steven J


    Despite wide consensus on needed changes in medical education, experts agree that the gap continues to widen between how physicians are trained and the future needs of our health care system. A new model for medical education is needed to create the medical school of the future. The American Medical Association (AMA) is working to support innovative models through partnerships with medical schools, educators, professional organizations, and accreditors. In 2013, the AMA designed an initiative to support rapid innovation among medical schools and disseminate the ideas being tested to additional medical schools. Awards of $1 million were made to 11 medical schools to redesign curricula for flexible, individualized learning pathways, measure achievement of competencies, develop new assessment tools to test readiness for residency, and implement new models for clinical experiences within health care systems. The medical schools have partnered with the AMA to create the AMA Accelerating Change in Medical Education Consortium, working together to share prototypes and participate in a national evaluation plan. Most of the schools have embarked on major curriculum revisions, replacing as much as 25% of the curriculum with new content in health care delivery and health system science in all four years of training. Schools are developing new certification in quality and patient safety and population management. In 2015, the AMA invited 21 additional schools to join the 11 founding schools in testing and disseminating innovation through the consortium and beyond.

  13. Creating Sister Cities: An Exchange Across Hemispheres (United States)

    Adams, M. T.; Cabezon, S. A.; Hardy, E.; Harrison, R. J.


    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.

  14. Cell adhesion pattern created by OSTE polymers. (United States)

    Liu, Wenjia; Li, Yiyang; Ding, Xianting


    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.

  15. Creating a culture where employee engagement Thrives

    Energy Technology Data Exchange (ETDEWEB)

    Don Groover, C.S.P. [Behavioral Science Technology, Ojai, CA (United States)


    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.


    Directory of Open Access Journals (Sweden)

    Anca-Simona HROMEI


    Full Text Available The realities of our modern society demonstrate that businesses operate in a competitive environment, which requires continuous improvement and development of production and sales. In this context, for financial, economic and fiscal reasons, companies resort to a process of concentration, through the merger of all the factors involved in the fulfilment and diversification of their activities. Often, the reasons for engaging in a merger refer to the fact that two merged companies create, in terms of value, an entity that exceeds the individual values of the two participants if they were to continue to activate separately. This paper aims at analysing whether the well-known relationship ‘1+1=3’, which describes merger synergies, applies at the Romanian level. For this reason, each of the values of 77 merged companies were compared with the sum of the individual values of the firms that initially created them. It was found that 56% of the mergers analysed generated added value for shareholders.

  17. Our Lunar Destiny: Creating a Lunar Economy (United States)

    Rohwer, Christopher J.


    "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.

  18. Creating quality improvement culture in public health agencies

    National Research Council Canada - National Science Library

    Davis, Mary V; Mahanna, Elizabeth; Joly, Brenda; Zelek, Michael; Riley, William; Verma, Pooja; Fisher, Jessica Solomon


    ...), or creating a quality improvement culture (n = 4). Agencies conducting formal quality improvement and creating a quality improvement culture had leadership support for quality improvement, participated in national...

  19. How to create high-performing teams. (United States)

    Lam, Samuel M


    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.

  20. Creating marketing strategies for higher education institutions

    Directory of Open Access Journals (Sweden)

    Lidia Białoń


    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.

  1. Creating Effective Dialogue Around Climate Change (United States)

    Kiehl, J. T.


    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.

  2. Creating Visual Design and Meaningful Audience Experiences

    DEFF Research Database (Denmark)

    Steijn, Arthur; Ion Wille, Jakob


    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......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...... 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...

  3. Creating Inclusive Youth Programs for LGBTQ+ Communities

    Directory of Open Access Journals (Sweden)

    Katherine E. Soule


    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.

  4. FoilSim: Basic Aerodynamics Software Created (United States)

    Peterson, Ruth A.


    FoilSim is interactive software that simulates the airflow around various shapes of airfoils. The graphical user interface, which looks more like a video game than a learning tool, captures and holds the students interest. The software is a product of NASA Lewis Research Center s Learning Technologies Project, an educational outreach initiative within the High Performance Computing and Communications Program (HPCCP).This airfoil view panel is a simulated view of a wing being tested in a wind tunnel. As students create new wing shapes by moving slider controls that change parameters, the software calculates their lift. FoilSim also displays plots of pressure or airspeed above and below the airfoil surface.

  5. Co-Creating ‘Second Life’

    DEFF Research Database (Denmark)

    Gürsimsek, Remzi Ates


    Developments in digital communication technologies, emergence of Social Media and shifting of digital media landscape towards a more participatory platform are not only the driving forces behind the implication of new technologies to the market, but they also have significant effects on the ways...... that mediate communication in Second Life (SL). The outlined theoretical framework and methodological approach is intended to summarize impressions from my observations of Second Life ‘builders’ in order to understand who they are, how they collaborate, and how they make sense of their co-design experiences...... people communicate, interact, participate and create communicative content in social contexts. This PhD project aims to observe how residents of Second Life use the Virtual World as a collaborative tool for creativity and co-design of the world’s content; that is, virtual places and artifacts...

  6. Creating biological nanomaterials using synthetic biology. (United States)

    Rice, MaryJoe K; Ruder, Warren C


    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.

  7. Creating a turnkey hierarchical geospatial data warehouse (United States)

    Libbey, Clinton R.


    The components of a web based data warehouse capable of populating, storing, searching, previewing, and disseminating a variety of information products over a TCP/IP network can be achieved through the use of commercially available products. The ability to deploy a solution that provides a central archive as well as distributed local archives of products is desirable. This approach is necessary due to the amount of data products generated for the community as well as the generation of custom products by regional users and the subsequent creation of their own archive for such products. Users must be able to leverage a central archive as well as have the ability to create their own local archive of data. This will increase efficiency at multiple levels and provide users with a solution that satisfies both their global data access needs and local data management.

  8. Minkowski metrics in creating universal ranking algorithms

    Directory of Open Access Journals (Sweden)

    Andrzej Ameljańczyk


    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

  9. Enhancing the Predictive Power of Mutations in the C-Terminus of the KCNQ1-Encoded Kv7.1 Voltage-Gated Potassium Channel

    NARCIS (Netherlands)

    Kapplinger, Jamie D.; Tseng, Andrew S.; Salisbury, Benjamin A.; Tester, David J.; Callis, Thomas E.; Alders, Marielle; Wilde, Arthur A. M.; Ackerman, Michael J.


    Despite the overrepresentation of Kv7.1 mutations among patients with a robust diagnosis of long QT syndrome (LQTS), a background rate of innocuous Kv7.1 missense variants observed in healthy controls creates ambiguity in the interpretation of LQTS genetic test results. A recent study showed that

  10. Enabling Reanalysis Intercomparison with the CREATE-IP and CREATE-V Projects (United States)

    Carriere, L.; Potter, G. L.; Hertz, J.; Shen, Y.; Britzolakis, G.; Peters, J.; Maxwell, T. P.; Li, J.; Strong, S.; Schnase, J. L.


    NASA Goddard Space Flight Center's Office of Computational and Information Sciences and Technology, the NASA Center for Climate Simulation (NCCS), 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. For CREATE-IP, our target reanalyses include ECMWF ERA-Interim, NASA/GMAO MERRA and MERRA2, NOAA/NCEP CFSR, NOAA/ESRL 20CR and 20CRv2, JMA JRA25, and JRA55. Each dataset is reformatted similarly to the models in the CMIP5 archive. By repackaging the reanalysis data into a common structure and format, it simplifies access, subsetting, and reanalysis comparison. Both monthly average data and a selection of high frequency data (6-hr) relevant to investigations such as the 2016 El Niño are provided. Much of the processing workflow has been automated and new data appear on a regular basis. In collaboration with the CLIVAR Global Synthesis and Observations Panel (GSOP), we are also processing and publishing eight ocean reanalyses, from 1980 to the present. Here, the data are regridded to a common 1° x 1° grid, vertically interpolated to the World Ocean Atlas 09 (WOA09) depths, and an ensemble is generated. CREATE-V is a web based visualization tool that allows the user to simultaneously view four reanalyses to facilitate comparison. The addition of a backend analytics engine, based on UV-CDAT and Scala provides the ability to generate a time series and anomaly for any given location on a map. The system enables scientists to identify data of interest and visualize, subset, and compare data without the need for download large volumes of data for local visualization.

  11. The pro-fusion domain of herpes simplex virus glycoprotein D (gD) interacts with the gD N terminus and is displaced by soluble forms of viral receptors. (United States)

    Fusco, Daniela; Forghieri, Cristina; Campadelli-Fiume, Gabriella


    Entry of herpes simplex virus into the cell requires the interaction of gD with one of its receptors, herpesvirus entry mediator or nectin 1, and the intervention of gB, gH, or gL, required to execute fusion of the virion envelope with cell membranes. The gD ectodomain is organized in two structurally and functionally differentiated regions. The N terminus (residues 1-260) carries the receptor binding sites, and the C terminus (residues 260-310) functions as the pro-fusion domain (PFD), which is required for viral infectivity and fusion but not for receptor binding. The objective of our studies is to elucidate how gD links receptor recognition to the triggering of fusion. Here, we show that PFD is made of subdomains 1 and 2 (amino acids 260-285 and 285-310). Each one partially contributed to herpes simplex virus infectivity. By means of glutathione S-transferase (GST) fusion proteins, we show that PFD bound soluble forms of gD, truncated at residue 260 (gD260t) or downstream. Both PFD subdomains bound gD260t, highlighting multiple contact sites between the N and C termini of gD. When gD260t was in complex with either receptor, it failed to bind GST-PFD. In turn, the receptors did not bind GST-PFD, irrespective of whether they were in complex with gD. Thus, gD260t interacted with the C terminus only if unbound to the receptor. We propose that (i) before receptor binding, gD adopts a "closed" conformation in which the N and C termini interact; and (ii) on encounter with a receptor, gD modifies its conformation and the N and C termini are released from reciprocal interactions ("opened" conformation) and enabled to trigger fusion.

  12. Creating an organizational culture for medication safety. (United States)

    Dennison, Robin Donohoe


    Medication errors are costly from human, economic, and societal perspectives. All patients are vulnerable to the detrimental effects of these errors. Recommendations regarding the problem of medication errors include: Prevention of error by learning from the nonpunitive reporting of errors and near misses; Evaluation of the system for potential causes of error through failure mode and effects analysis and encouragement of a questioning attitude; Elimination of system problems that increase the risk of error; Recognition that humans are fallible and that error will occur even in a perfect system; Minimization of the consequences of errors when they do occur. An important goal for healthcare organizations should be to create a culture that accepts the imperfection of human performance and solicits the assistance of team members in the development of safeguards for error prevention. Proposed interventions to prevent medication errors can be described by the PATIENT SAFE taxonomy, which includes: Patient participation; Adherence to established policy and procedures; Technology use; Information accessibility; Education regarding medication safety; Nonpunitive approach to reporting of errors and near misses; Teamwork, communication, and collaboration; Staffing: adequate number and staffing mix; Administration support for the clinical goal of patient safety; Failure mode and effects analysis with team member involvement; Environment and equipment to support patient safety

  13. Creating Affording Situations: Coaching through Animate Objects. (United States)

    Baber, Chris; Khattab, Ahmad; Russell, Martin; Hermsdörfer, Joachim; Wing, Alan


    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.

  14. Creating Affording Situations: Coaching through Animate Objects

    Directory of Open Access Journals (Sweden)

    Chris Baber


    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.

  15. Informational conflicts created by the waggle dance (United States)

    Grüter, Christoph; Balbuena, M. Sol; Farina, Walter M


    The honeybee (Apis mellifera) waggle dance is one of the most intriguing animal communication signals. A dancing bee communicates the location of a profitable food source and its odour. Followers may often experience situations in which dancers indicate an unfamiliar location but carry the scent of a flower species the followers experienced previously at different locations. Food scents often reactivate bees to resume food collection at previously visited food patches. This double function of the dance creates a conflict between the social vector information and the private navigational information. We investigated which kind of information followers with field experience use in this situation and found that followers usually ignored the spatial information encoded by the waggle dance even if they followed a dance thoroughly (five waggle runs or more). They relied on private information about food source locations instead (in 93% of all cases). Furthermore, foragers preferred to follow dancers carrying food odours they knew from previous field trips, independently of the spatial information encoded in the dance. Surprisingly, neither odour identity nor the location indicated by the dancer was an important factor for the reactivation success of a dance. Our results contrast with the assumption that (i) followers usually try to decode the vector information and (ii) dances indicating an unfamiliar location are of little interest to experienced foragers. PMID:18331980

  16. Creating the Nurses' Environmental Awareness Tool (NEAT). (United States)

    Schenk, Elizabeth; Butterfield, Patricia; Postma, Julie; Barbosa-Leiker, Celestina; Corbett, Cindy


    Acute care delivery creates secondary health risks to patients, health care workers, and the environment through a complex waste stream, intensive energy use, and frequent use of harmful chemicals. Nurses are among the most affected by these risks and are also pivotal change agents in reducing the negative impacts of health care delivery. Assessing nurses' understanding of health care-associated environmental health risks is essential if care is to be delivered in an environmentally safe and healthy manner, as indicated by published professional standards of nursing practice. However, psychometrically sound instruments that measure nurses' awareness of the environmental impacts of nursing practice are not available. To address this gap, a prototype of the Nurse's Environmental Awareness Tool (NEAT) was developed. Seven content experts in environmental health nursing and/or psychometrics were asked to review draft items. Comments were analyzed and applied to the scale items. Several revisions from the original item pool were made. The resulting draft NEAT includes six subscales, in three paired subsets. This article provides a summary of the process of item development and scale design. These findings, although preliminary, provide a foundation for subsequent psychometric testing. The result of this study is the creation of an instrument to measure nurses' awareness of and behaviors associated with the environmental impact of their practices. © 2015 The Author(s).

  17. Creating the optimal conditions for rehabilitation research. (United States)

    Bakheit, Abdel Magid


    Current evidence suggests that productivity in biomedical research depends on the support that the research group enjoys from its parent institution, the composition of the group and the personal attributes of its members and leader. Supportive institutions provide adequate physical resources and allow substantial uninterrupted time for research. The effectiveness of the research group asa whole is also strongly influenced by the group's structure, the professional competence of the group leader, his leadership style and his ability to foster collaboration with other research groups and organizations. There is a good case for a flexible leadership style that is modelled on the situational theory of leadership. In addition, the personal characteristics of the individual members of the research group influence the quality and quantity of the research output. Effective groups are made of motivated individuals with research training and skills that are relevant to the objectives of the research group. Rehabilitation research is fundamentally different from traditional biomedical research. This study discusses how the factors that influence productivity of biomedical research relate to rehabilitation science and practice and examines the conditions that are necessary to create and maintain an academic environment that is conducive to large volume,high-quality research in rehabilitation medicine.

  18. Creating a Universe, a Conceptual Model

    Directory of Open Access Journals (Sweden)

    James R. Johnson


    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.

  19. Creating the optimized international marketing mix

    Directory of Open Access Journals (Sweden)

    Grubor Aleksandar


    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.

  20. Creating healthy work environments: a strategic perspective. (United States)

    Adamson, Bonnie J


    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.

  1. Creating a national home visiting research network. (United States)

    Duggan, Anne; Minkovitz, Cynthia S; Chaffin, Mark; Korfmacher, Jon; Brooks-Gunn, Jeanne; Crowne, Sarah; Filene, Jill; Gonsalves, Kay; Landsverk, John; Harwood, Robin


    Home visiting can play a key role in the early childhood system of services. For home visiting to achieve its potential, decision-makers must make informed choices regarding adoption, adaptation, coordination, scale-up, and sustainment. We need a coordinated, focused, and theory-based home visiting research infrastructure to inform such decisions. The transdisciplinary Home Visiting Research Network (HVRN) was established in July 2012 with funding from the Health Resources and Services Administration. Its goal is to promote the translation of research findings into policy and practice. Its objectives are to (1) develop a national home visiting research agenda, (2) advance the use of innovative research methods; and (3) provide a research environment that is supportive of the professional development of emerging researchers interested in home visiting. A Management Team designs and directs activities to achieve these objectives through Work Teams. A Steering Committee of national leaders representing stakeholder groups oversees progress. HVRN's Coordinating Center supports the Work Teams and HVRN's Home visiting Applied Research Collaborative, a practice-based research network of home visiting programs. This article describes HVRN's rationale, approach, and anticipated products. We use home visiting-primary care coordination as an illustration, noting potential roles for pediatric practices and pediatric researchers and research educators in HVRN activities. HVRN creates the infrastructure for a rigorous program of research to inform policy and practice on home visiting as part of the system of services to improve family functioning, parenting, and child outcomes.

  2. Creating better superconductors by periodic nanopatterning

    Directory of Open Access Journals (Sweden)

    Milan P. Allan, Mark H. Fischer, Oliver Ostojic, Arjo Andringa


    Full Text Available The quest to create superconductors with higher transition temperatures is as old as superconductivity itself. One strategy, popular after the realization that (conventional superconductivity is mediated by phonons, is to chemically combine different elements within the crystalline unit cell to maximize the electron-phonon coupling. This led to the discovery of NbTi and Nb3Sn, to name just the most technologically relevant examples. Here, we propose a radically different approach to transform a `pristine' material into a better (meta- superconductor by making use of modern fabrication techniques: designing and engineering the electronic properties of thin films via periodic patterning on the nanoscale. We present a model calculation to explore the key effects of different supercells that could be fabricated using nanofabrication or deliberate lattice mismatch, and demonstrate that specific pattern will enhance the coupling and the transition temperature. We also discuss how numerical methods could predict the correct design parameters to improve superconductivity in materials including Al, NbTi, and MgB2

  3. Dancing Lights: Creating the Aurora Story (United States)

    Wood, E. L.; Cobabe-Ammann, E. A.


    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.

  4. Creating cultures of excellence: Strategies and outcomes

    Directory of Open Access Journals (Sweden)

    Michael Mintrom


    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.

  5. Creating conditions for cooperative learning: Basic elements

    Directory of Open Access Journals (Sweden)

    Ševkušić-Mandić Slavica G.


    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.

  6. Creating Web Services from Community Sourced Data (United States)

    Siegel, D.; Scopel, C.; Boghici, E.


    In order to extend the World Hydro Basemap and build watershed delineation and river tracing services that cover the entire planet, we are integrating community-contributed data into a global hydrographic dataset. This dataset is the engine behind a foundational set of tools and services intended to enable hydrologic analysis on the web. However, each organization that collects hydrography uses a workflow and data model unique to their mission, which makes synthesizing their data difficult. Furthermore, these data are collected at different resolutions, so running analytics across regions with multiple contributors is not necessarily valid. Thus, instead of merging contributed data into a seamless geodatabase, the goal of our Community Maps for Hydrology program is to create workflows for converting any arbitrary dataset into the Arc Hydro Data Model. This way, tools and services can be pointed towards different contributions interchangeably while still maintaining the autonomy of each dataset. Contributors retain ownership of their data and are responsible for updates and edits, but the tools and services work identically across all contributions. HydroSHEDs data, contributed by the World Wildlife Fund, is used at the smallest scales to ensure global coverage, and national datasets extend our services to the medium-scales where available. A workflow to incorporate LIDAR and other large scale data is being developed as well, so that local governments and engineering companies can contribute to the program. Watershed Delineation Tool The World Hydro Basemap

  7. Creating supportive environments for AIDS prevention. (United States)


    The World Health Organization's Global Programme on AIDS (GPA) argues that AIDS prevention requires a supportive environment, but that discriminatory laws make marginalized people even more vulnerable to sexually transmitted diseases (STDs). A country's legal, economic, and social environments can influence the pandemic. A short-term measure could be prostitutes collectively insisting that their clients wear condoms. Long-term measures of AIDS prevention require the improvement of the legal status of women and their access to education. Societies repress or tolerate drug use, prostitution, homosexuality, and casual sex, but often ministries forbid condom advertising, and condom possession by women can be used as evidence of prostitution. Fear of mandatory testing and detention prevents sex workers and drug users from accepting condoms and needles. A recent review of policies in 22 locations around the world found low seroprevalence of HIV among IV drug users in only 5 countries -- exactly the same countries where IV drug users had legal access to sterile needles. In Zambia a national condom promotion campaign was launched only after a 2-year debate, while free condoms had been distributed surreptitiously by a nongovernmental organization. Sex discrimination in many countries forces women to trade sex for money to make a living, and women in sex work are very vulnerable to HIV infection. Overcoming the subordination of women is a long-term undertaking, but an example of successful short-term empowerment of women is a credit scheme operated by a bank for rural women in Bangladesh. Socially, culturally, and economically male infidelity is often condoned, creating the risk of HIV infection and of passing the infection on to wives. Information campaigns stressing shared responsibility can be effective in changing social norms. Some traditional practices, e.g., ritual cleansing in Uganda and Zambia, also expose participants to the risk of HIV infection.


    Directory of Open Access Journals (Sweden)

    Gauca Oana


    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.

  9. Defectors Can Create Conditions That Rescue Cooperation.

    Directory of Open Access Journals (Sweden)

    Adam James Waite


    Full Text Available Cooperation based on the production of costly common goods is observed throughout nature. This is puzzling, as cooperation is vulnerable to exploitation by defectors which enjoy a fitness advantage by consuming the common good without contributing fairly. Depletion of the common good can lead to population collapse and the destruction of cooperation. However, population collapse implies small population size, which, in a structured population, is known to favor cooperation. This happens because small population size increases variability in cooperator frequency across different locations. Since individuals in cooperator-dominated locations (which are most likely cooperators will grow more than those in defector-dominated locations (which are most likely defectors, cooperators can outgrow defectors globally despite defectors outgrowing cooperators in each location. This raises the possibility that defectors can lead to conditions that sometimes rescue cooperation from defector-induced destruction. We demonstrate multiple mechanisms through which this can occur, using an individual-based approach to model stochastic birth, death, migration, and mutation events. First, during defector-induced population collapse, defectors occasionally go extinct before cooperators by chance, which allows cooperators to grow. Second, empty locations, either preexisting or created by defector-induced population extinction, can favor cooperation because they allow cooperator but not defector migrants to grow. These factors lead to the counterintuitive result that the initial presence of defectors sometimes allows better survival of cooperation compared to when defectors are initially absent. Finally, we find that resource limitation, inducible by defectors, can select for mutations adaptive to resource limitation. When these mutations are initially present at low levels or continuously generated at a moderate rate, they can favor cooperation by further reducing local

  10. Creating Porcine Biomedical Models Through Recombineering

    Directory of Open Access Journals (Sweden)

    Lawrence B. Schook


    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.

  11. Peculiarities of Creating Foreign Students’ Lexical Competence

    Directory of Open Access Journals (Sweden)

    Aigul Eskermesovna Sadenova


    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

  12. Effects of systematic N-terminus deletions and benzoylations of endogenous RF-amide peptides on NPFF1R, NPFF2R, GPR10, GPR54 and GPR103. (United States)

    Rouméas, Laurent; Humbert, Jean-Paul; Schneider, Séverine; Doebelin, Christelle; Bertin, Isabelle; Schmitt, Martine; Bourguignon, Jean-Jacques; Simonin, Frédéric; Bihel, Frédéric


    Mammalian RF-amide peptides including RF-amide-related peptides-1 and -3, neuropeptides AF and FF, Prolactin releasing peptides, Kisspeptins and RFa peptides are currently considered endogenous peptides for the GPCRs NPFF1R, NPFF2R, GPR10, GPR54 and GPR103, respectively. While NPFF1R and NPFF2R displayed high affinity for all the RF-amide peptides, GPR10, GPR54 and GPR103 only bind their cognate ligands. Through a systematic and sequential N-terminus deletion and benzoylation of either RF-amide neuropeptide (RFRP-3, NPFF, Kp-10, PrRP20, and 26RFa), we report the corresponding impact on affinity and activity towards all the RF-amide receptors (NPFF1R, NPFF2R, GPR10, GPR54 and GPR103). Our results highlight the difficulty to develop selective peptide ligands for GPR10, GPR54 or GPR103 without a modification of the C-terminus RF-amide signature, but open the door to the design of new RF-amide peptides acting as agonist for one receptor and antagonist for another one. Copyright © 2015 Elsevier Inc. All rights reserved.

  13. Heparin Oligosaccharides Inhibit Chemokine (CXC Motif) Ligand 12 (CXCL12) Cardioprotection by Binding Orthogonal to the Dimerization Interface, Promoting Oligomerization, and Competing with the Chemokine (CXC Motif) Receptor 4 (CXCR4) N Terminus* (United States)

    Ziarek, Joshua J.; Veldkamp, Christopher T.; Zhang, Fuming; Murray, Nathan J.; Kartz, Gabriella A.; Liang, Xinle; Su, Jidong; Baker, John E.; Linhardt, Robert J.; Volkman, Brian F.


    The ability to interact with cell surface glycosaminoglycans (GAGs) is essential to the cell migration properties of chemokines, but association with soluble GAGs induces the oligomerization of most chemokines including CXCL12. Monomeric CXCL12, but not dimeric CXCL12, is cardioprotective in a number of experimental models of cardiac ischemia. We found that co-administration of heparin, a common treatment for myocardial infarction, abrogated the protective effect of CXCL12 in an ex vivo rat heart model for myocardial infarction. The interaction between CXCL12 and heparin oligosaccharides has previously been analyzed through mutagenesis, in vitro binding assays, and molecular modeling. However, complications from heparin-induced CXCL12 oligomerization and studies using very short oligosaccharides have led to inconsistent conclusions as to the residues involved, the orientation of the binding site, and whether it overlaps with the CXCR4 N-terminal site. We used a constitutively dimeric variant to simplify the NMR analysis of CXCL12-binding heparin oligosaccharides of varying length. Biophysical and mutagenic analyses reveal a CXCL12/heparin interaction surface that lies perpendicular to the dimer interface, does not involve the chemokine N terminus, and partially overlaps with the CXCR4-binding site. We further demonstrate that heparin-mediated enzymatic protection results from the promotion of dimerization rather than direct heparin binding to the CXCL12 N terminus. These results clarify the structural basis for GAG recognition by CXCL12 and lend insight into the development of CXCL12-based therapeutics. PMID:23148226

  14. 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 ( 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.

  15. Palmitoylation at two cysteine clusters on the C-terminus of GluN2A and GluN2B differentially control synaptic targeting of NMDA receptors.

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    Hayley A Mattison

    Full Text Available Palmitoylation of NMDARs occurs at two distinct cysteine clusters in the carboxyl-terminus of GluN2A and GluN2B subunits that differentially regulates retention in the Golgi apparatus and surface expression of NMDARs. Mutations of palmitoylatable cysteine residues in the membrane-proximal cluster to non-palmitoylatable serines leads to a reduction in the surface expression of recombinant NMDARs via enhanced internalization of the receptors. Mutations in a cluster of cysteines in the middle of the carboxyl-terminus of GluN2A and GluN2B, leads to an increase in the surface expression of NMDARs via an increase in post-Golgi trafficking. Using a quantitative electrophysiological assay, we investigated whether palmitoylation of GluN2 subunits and the differential regulation of surface expression affect functional synaptic incorporation of NMDARs. We show that a reduction in surface expression due to mutations in the membrane-proximal cluster translates to a reduction in synaptic expression of NMDARs. However, increased surface expression induced by mutations in the cluster of cysteines that regulates post-Golgi trafficking of NMDARs does not increase the synaptic pool of NMDA receptors, indicating that the number of synaptic receptors is tightly regulated.

  16. Creating a Servitude to solve an encroachment Dispute: A Solution or creating another Problem?

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    Zsa-Zsa Temmers Boggenpoel


    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

  17. Merging Galaxies Create a Binary Quasar (United States)


    Astronomers have found the first clear evidence of a binary quasar within a pair of actively merging galaxies. Quasars are the extremely bright centers of galaxies surrounding super-massive black holes, and binary quasars are pairs of quasars bound together by gravity. Binary quasars, like other quasars, are thought to be the product of galaxy mergers. Until now, however, binary quasars have not been seen in galaxies that are unambiguously in the act of merging. But images of a new binary quasar from the Carnegie Institution's Magellan telescope in Chile show two distinct galaxies with "tails" produced by tidal forces from their mutual gravitational attraction. "This is really the first case in which you see two separate galaxies, both with quasars, that are clearly interacting," says Carnegie astronomer John Mulchaey who made observations crucial to understanding the galaxy merger. Most, if not all, large galaxies, such as our galaxy the Milky Way, host super-massive black holes at their centers. Because galaxies regularly interact and merge, astronomers have assumed that binary super-massive black holes have been common in the Universe, especially during its early history. Black holes can only be detected as quasars when they are actively accreting matter, a process that releases vast amounts of energy. A leading theory is that galaxy mergers trigger accretion, creating quasars in both galaxies. Because most such mergers would have happened in the distant past, binary quasars and their associated galaxies are very far away and therefore difficult for most telescopes to resolve. The binary quasar, labeled SDSS J1254+0846, was initially detected by the Sloan Digital Sky Survey, a large scale astronomical survey of galaxies and over 120,000 quasars. Further observations by Paul Green of the Harvard-Smithsonian Center for Astrophysics and colleagues* using NASA's Chandra's X-ray Observatory and telescopes at Kitt Peak National Observatory in Arizona and Palomar

  18. Creating Partnerships on Campus to Facilitate Practical Experiences (United States)

    Becker, Craig M.; Johnson, Hans; McNeil, Michael P.; Warren, Karen


    College campuses create small communities where mutually beneficial partnerships can be used to create practical work experiences for students. The procedure outlined in this article outlines how to create a partnership between the campus health and recreation center and an academic department to evaluate the implementation of a new smoking…

  19. Download - CREATE portal | LSDB Archive [Life Science Database Archive metadata

    Lifescience Database Archive (English)

    Full Text Available .6 KB) Simple search and download 3 InCeP (1 KB) Simple search and download 4 InCeP images (13.4 MB) - 5 KeyMolnet data (7.1 MB) - 6 Mascot

  20. 31 CFR 900.8 - No private rights created. (United States)


    ... 31 Money and Finance: Treasury 3 2010-07-01 2010-07-01 false No private rights created. 900.8... No private rights created. The standards in this chapter do not create any right or benefit, substantive or procedural, enforceable at law or in equity by a party against the United States, its agencies...

  1. 45 CFR 30.9 - No private rights created. (United States)


    ... 45 Public Welfare 1 2010-10-01 2010-10-01 false No private rights created. 30.9 Section 30.9... Provisions § 30.9 No private rights created. The standards in this part do not create any right or benefit, substantive or procedural, enforceable at law or in equity by a party against the United States, the...

  2. 10 CFR 1015.108 - No private rights created. (United States)


    ... 10 Energy 4 2010-01-01 2010-01-01 false No private rights created. 1015.108 Section 1015.108... § 1015.108 No private rights created. The standards in this part do not create any right or benefit, substantive or procedural, enforceable at law or in equity by a party against the United States, its agencies...

  3. Identification of a classic nuclear localization signal at the N terminus that regulates the subcellular localization of Rbfox2 isoforms during differentiation of NMuMG and P19 cells. (United States)

    Wenzel, Manuel; Schüle, Martin; Casanovas, Sonia; Strand, Dennis; Strand, Susanne; Winter, Jennifer


    Nuclear localization of the alternative splicing factor Rbfox2 is achieved by a C-terminal nuclear localization signal (NLS) which can be excluded from some Rbfox2 isoforms by alternative splicing. While this predicts nuclear and cytoplasmic localization, Rbfox2 is exclusively nuclear in some cell types. Here, we identify a second NLS in the N terminus of Rbfox2 isoform 1A that is not included in Rbfox2 isoform 1F. Rbfox2 1A isoforms lacking the C-terminal NLS are nuclear, whereas equivalent 1F isoforms are cytoplasmic. A shift in Rbfox2 expression toward cytoplasmic 1F isoforms occurs during epithelial to mesenchymal transition (EMT) and could be important in regulating the activity and function of Rbfox2. © 2016 Federation of European Biochemical Societies.

  4. Synthetic biology ethically evaluated: The creating God and co-creating human

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    Riaan Rheeder


    Full Text Available God did not create once and then put an end to it. Testimony from Scripture shows that God continuously establishes or creates new things. Humans can therefore expect to always see and experience new things in creation. With this pattern of reasoning, one can anticipate that the human being as image of God will continuously establish new things in history. Although nature has value, it does not have absolute value and therefore it can be synthesised responsibly. The thought that humans are stewards of God is no longer adequate to, theologically put into words, the relationship human beings have with nature. New biotechnological developments ask for different answers from Scripture. Several ethicists are of the opinion that the theological construction of humans and created co-creators can help found the relationship of the human being to nature. Humans developed as God’s image evolutionary. On the one hand, this means humans themselves are a product of nature. On the other hand, the fact that humans are the image of God is also an ethical call that humans, like God, have to develop and create new things throughout history. Synthetic biology can be evaluated as technology that is possible, because humans are the image of God. However, it should, without a doubt, be executed responsibly.Sintetiese biologie eties geëvalueer: Die skeppende God en medeskeppende mens. God het nie net eenmaal geskep en daar gestop nie. Uit Skrifgetuienisse kan afgelei word dat God voortdurend nuwe dinge tot stand bring of skep. Daarom kan die mens verwag om gedurig nuwe dinge in die skepping te sien en te beleef. Hiermee saam kan verwag word dat die mens as beeld van God voortdurend nuwe dinge in die geskiedenis tot stand sal bring. Alhoewel die natuur waarde het, het dit nie absolute waarde nie en kan dus verantwoordelik gesintetiseer word. Die gedagte dat die mens rentmeester van God is, is nie meer voldoende om die mens se verhouding tot die natuur teologies te

  5. Protein Aggregation Formed by Recombinant cp19k Homologue of Balanus albicostatus Combined with an 18 kDa N-Terminus Encoded by pET-32a(+) Plasmid Having Adhesion Strength Comparable to Several Commercial Glues. (United States)

    Liang, Chao; Li, Yunqiu; Liu, Zhiming; Wu, Wenjian; Hu, Biru


    The barnacle is well known for its tenacious and permanent attachment to a wide variety of underwater substrates, which is accomplished by synthesizing, secreting and curing a mixture of adhesive proteins termed "barnacle cement". In order to evaluate interfacial adhesion abilities of barnacle cement proteins, the cp19k homologous gene in Balanus albicostatus (Balcp19k) was cloned and expressed in Escherichia coli. Here, we report an intriguing discovery of a gel-like super adhesive aggregation produced by Trx-Balcp19k, a recombinant Balcp19k fusion protein. The Trx-Balcp19k consists of an 18 kDa fragment at the N-terminus, which is encoded by pET-32a(+) plasmid and mainly comprised of a thioredoxin (Trx) tag, and Balcp19k at the C-terminus. The sticky aggregation was designated as "Trx-Balcp19k gel", and the bulk adhesion strength, biochemical composition, as well as formation conditions were all carefully investigated. The Trx-Balcp19k gel exhibited strong adhesion strength of 2.10 ± 0.67 MPa, which was approximately fifty folds higher than that of the disaggregated Trx-Balcp19k (40 ± 8 kPa) and rivaled those of commercial polyvinyl acetate (PVA) craft glue (Mont Marte, Australia) and UHU glue (UHU GmbH & Co. KG, Germany). Lipids were absent from the Trx-Balcp19k gel and only a trace amount of carbohydrates was detected. We postulate that the electrostatic interactions play a key role in the formation of Trx-Balcp19k gel, by mediating self-aggregation of Trx-Balcp19k based on its asymmetric distribution pattern of charged amino acids. Taken together, we believe that our discovery not only presents a promising biological adhesive with potential applications in both biomedical and technical fields, but also provides valuable paradigms for molecular design of bio-inspired peptide- or protein-based materials.

  6. The C-terminal 20 Amino Acids of Drosophila Topoisomerase 2 Are Required for Binding to a BRCA1 C Terminus (BRCT) Domain-containing Protein, Mus101, and Fidelity of DNA Segregation* (United States)

    Chen, Yu-tsung Shane; Wu, Jianhong; Modrich, Paul; Hsieh, Tao-shih


    Eukaryotic topoisomerase 2 (Top2) and one of its interacting partners, topoisomerase IIβ binding protein 1 (TopBP1) are two proteins performing essential cellular functions. We mapped the interacting domains of these two proteins using co-immunoprecipitation and pulldown experiments with truncated or mutant Drosophila Top2 with various Ser-to-Ala substitutions. We discovered that the last 20 amino acids of Top2 represent the key region for binding with Mus101 (the Drosophila homolog of TopBP1) and that phosphorylation of Ser-1428 and Ser-1443 is important for Top2 to interact with the N terminus of Mus101, which contains the BRCT1/2 domains. The interaction between Mus101 and the Top2 C-terminal regulatory domain is phosphorylation-dependent because treatment with phosphatase abolishes their association in pulldown assays. The binding affinity of N-terminal Mus101 with a synthetic phosphorylated peptide spanning the last 25 amino acids of Top2 (with Ser(P)-1428 and Ser(P)-1443) was determined by surface plasmon resonance with a Kd of 0.57 μm. In an in vitro decatenation assay, Mus101 can specifically reduce the decatenation activity of Top2, and dephosphorylation of Top2 attenuates this response. Next, we endeavored to establish a cellular system for testing the biological function of Top2-Mus101 interaction. Top2-silenced S2 cells rescued by Top2Δ20, Top2 with 20 amino acids truncated from the C terminus, developed abnormally high chromosome numbers, which implies that Top2-Mus101 interaction is important for maintaining the fidelity of chromosome segregation during mitosis. PMID:27129233

  7. Disruption of hydrogen bonds between major histocompatibility complex class II and the peptide N-terminus is not sufficient to form a human leukocyte antigen-DM receptive state of major histocompatibility complex class II. (United States)

    Schulze, Monika-Sarah E D; Anders, Anne-Kathrin; Sethi, Dhruv K; Call, Melissa J


    Peptide presentation by MHC class II is of critical importance to the function of CD4+ T cells. HLA-DM resides in the endosomal pathway and edits the peptide repertoire of newly synthesized MHC class II molecules before they are exported to the cell surface. HLA-DM ensures MHC class II molecules bind high affinity peptides by targeting unstable MHC class II:peptide complexes for peptide exchange. Research over the past decade has implicated the peptide N-terminus in modulating the ability of HLA-DM to target a given MHC class II:peptide combination. In particular, attention has been focused on both the hydrogen bonds between MHC class II and peptide, and the occupancy of the P1 anchor pocket. We sought to solve the crystal structure of a HLA-DR1 molecule containing a truncated hemagglutinin peptide missing three N-terminal residues compared to the full-length sequence (residues 306-318) to determine the nature of the MHC class II:peptide species that binds HLA-DM. Here we present structural evidence that HLA-DR1 that is loaded with a peptide truncated to the P1 anchor residue such that it cannot make select hydrogen bonds with the peptide N-terminus, adopts the same conformation as molecules loaded with full-length peptide. HLA-DR1:peptide combinations that were unable to engage up to four key hydrogen bonds were also unable to bind HLA-DM, while those truncated to the P2 residue bound well. These results indicate that the conformational changes in MHC class II molecules that are recognized by HLA-DM occur after disengagement of the P1 anchor residue.

  8. An alpha-helical domain within the carboxyl terminus of herpes simplex virus type 1 (HSV-1) glycoprotein B (gB) is associated with cell fusion and resistance to heparin inhibition of cell fusion. (United States)

    Foster, T P; Melancon, J M; Kousoulas, K G


    Previous studies from our laboratory indicated that a 28-amino-acid carboxyl-terminal truncation of gB caused extensive virus-induced cell fusion (Baghian et al., 1993, J Virol 67, 2396-2401). We tested the ability of additional truncations and mutations within gB to cause cell fusion in the recently established virus-free cell fusion assay (Turner et al., 1998, J. Virol. 72, 873-875). Deletion of the carboxyl-terminal 28 amino acids of gB (gBDelta28), which removed part of the predicted alpha-helical structure H17b, caused extensive cell fusion. A gB truncation specified by gBDelta36, which removed the entire H17b domain, caused as much cell fusion as the gBDelta28 truncation. Similarly, gB(A874P) containing a substitution of an Ala with Pro within H17b caused cell fusion. Heparin, a gB-specific inhibitor of virus-induced cell fusion, inhibited both wild-type gB and gB(syn3)-mediated cell fusion. In contrast, fusion of cells transfected with gB(Delta28), gB(Delta36), or gB(A874P) was resistant to heparin inhibition of cell fusion. We concluded the following: (1) The predicted alpha-helical structure of H17b within the carboxyl terminus of gB is involved in both virus-induced and virus-free cell fusion. (2) Heparin is a specific inhibitor of gB-mediated fusion in both systems. (3) Resistance to heparin inhibition of gB-mediated cell fusion is associated with the predicted alpha-helical structure H17b within the carboxyl terminus of gB. Copyright 2001 Academic Press.

  9. Creating real network with expected degree distribution: A statistical simulation


    WenJun Zhang; GuangHua Liu


    The degree distribution of known networks is one of the focuses in network analysis. However, its inverse problem, i.e., to create network from known degree distribution has not yet been reported. In present study, a statistical simulation algorithm was developed to create real network with expected degree distribution. It is aniteration procedure in which a real network, with the least deviation of actual degree distribution to expected degree distribution, was created. Random assignment was...



    ENGİNOĞLU, Didem; ARIKAN, Cenk Laçin


    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 ...

  11. Creating 3D gelatin phantoms for experimental evaluation in biomedicine

    Directory of Open Access Journals (Sweden)

    Stein Nils


    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.

  12. 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

  13. Mandatory Parent Education Programs Can Create Positive Youth Sport Experiences (United States)

    Christofferson, Jennifer; Strand, Bradford


    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…

  14. Creating a Culture of Peace in the Elementary Classroom (United States)

    Hunter, Tiffany J.


    In this article, the author shares how she created a "peaceable classroom" through activities which she incorporated into a 1st-grade curriculum and which fulfilled academic requirements. As a 1st-grade teacher at Redlands Adventist Academy in Redlands, CA, she wanted to create a learning environment that would foster values like…

  15. Creating Cartoons to Promote Leaderships Skills and Explore Leadership Qualities (United States)

    Smith, Latisha L.; Clausen, Courtney K.; Teske, Jolene K.; Ghayoorrad, Maryam; Gray, Phyllis; Al Subia, Sukainah; Atwood-Blaine, Dana; Rule, Audrey C.


    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…

  16. Investigations into properties of charge traps created in CCDs by ...

    Indian Academy of Sciences (India)

    Charge traps we observe are created when a vacancy in the crystalline structure of silicon (mobile by itself) combines with another vacancy (V) or doping atom (P) or impurity atom (O) to create a complex. Such a complex (VV or VP or VO) is immobile and has the ability to capture and retain electrons from charge packets.

  17. 5 CFR 9701.372 - Creating initial pay ranges. (United States)


    ... 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...

  18. 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 ...

  19. Fundamental Characteristics of Incentive Streams Created by Legal Systems

    NARCIS (Netherlands)

    Dari Mattiacci, G.


    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

  20. Creating Learning Outcomes for a TESOL Teacher Preparation Program (United States)

    Henrichsen, Lynn; Tanner, Mark


    This article shares the results of a multisemester effort to create learning outcomes and related assessment measures for a graduate-level teaching English to speakers of other languages (TESOL) teacher preparation program. It starts by explaining what learning outcomes are, why they are important, and how to create them. It then describes the…