Membrane fission via transmembrane contact.

Division of intracellular organelles often correlates with additional membrane wrapping, e.g., by the endoplasmic reticulum or the outer mitochondrial membrane. Such wrapping plays a vital role in proteome and lipidome organization. However, how an extra membrane impacts the mechanics of the division has not been investigated. Here we combine fluorescence and cryo-electron microscopy experiments with self-consistent field theory to explore the stress-induced instabilities imposed by membrane wrapping in a simple double-membrane tubular system. We find that, at physiologically relevant conditions, the outer membrane facilitates an alternative pathway for the inner-tube fission through the formation of a transient contact (hemi-fusion) between both membranes. A detailed molecular theory of the fission pathways in the double membrane system reveals the topological complexity of the process, resulting both in leaky and leakless intermediates, with energies and topologies predicting physiological events.

Enriched Environment Contributes to the Recovery from Neurotoxin-Induced Parkinson's Disease Pathology.

Parkinson's disease (PD) is a neurological disorder that affects dopaminergic neurons. The lack of understanding of the underlying molecular mechanisms of PD pathology makes treating it a challenge. Several pieces of evidence support the protective role of enriched environment (EE) and exercise on dopaminergic neurons. The specific aspect(s) of neuroprotection after exposure to EE have not been identified. Therefore, we have investigated the protective role of EE on dopamine dysregulation and subsequent downregulation of DJ1 protein using in vitro and in vivo models of PD. Our study for the first time demonstrated that DJ1 expression has a direct correlation with dopamine downregulation in PD models and exposure to EE has a significant impact on improving the behavioral changes in PD mice. This research provides evidence that exercise in EE has a positive effect on PD without interfering with the current line of therapy.

Allosteric control of dynamin-related protein 1 through a disordered C-terminal Short Linear Motif.

The mechanochemical GTPase dynamin-related protein 1 (Drp1) catalyzes mitochondrial and peroxisomal fission, but the regulatory mechanisms remain ambiguous. Here we find that a conserved, intrinsically disordered, six-residue Short Linear Motif at the extreme Drp1 C-terminus, named CT-SLiM, constitutes a critical allosteric site that controls Drp1 structure and function in vitro and in vivo. Extension of the CT-SLiM by non-native residues, or its interaction with the protein partner GIPC-1, constrains Drp1 subunit conformational dynamics, alters self-assembly properties, and limits cooperative GTP hydrolysis, surprisingly leading to the fission of model membranes in vitro. In vivo, the involvement of the native CT-SLiM is critical for productive mitochondrial and peroxisomal fission, as both deletion and non-native extension of the CT-SLiM severely impair their progression. Thus, contrary to prevailing models, Drp1-catalyzed membrane fission relies on allosteric communication mediated by the CT-SLiM, deceleration of GTPase activity, and coupled changes in subunit architecture and assembly-disassembly dynamics.

Allosteric control of dynamin-related protein 1-catalyzed mitochondrial fission through a conserved disordered C-terminal Short Linear Motif.

The mechanochemical GTPase dynamin-related protein 1 (Drp1) catalyzes mitochondrial fission, but the regulatory mechanisms remain ambiguous. Here we found that a conserved, intrinsically disordered, six-residue Short Linear Motif at the extreme Drp1 C-terminus, named CT-SLiM, constitutes a critical allosteric site that controls Drp1 structure and function in vitro and in vivo. Extension of the CT-SLiM by non-native residues, or its interaction with the protein partner GIPC-1, constrains Drp1 subunit conformational dynamics, alters self-assembly properties, and limits cooperative GTP hydrolysis, leading to the fission of model membranes in vitro. In vivo, the availability of the native CT-SLiM is a requirement for productive mitochondrial fission, as both non-native extension and deletion of the CT-SLiM severely impair its progression. Thus, contrary to prevailing models, Drp1-catalyzed mitochondrial fission relies on allosteric communication mediated by the CT-SLiM, deceleration of GTPase activity, and coupled changes in subunit architecture and assembly-disassembly dynamics.

D1 and D2 neurons in the nucleus accumbens enable positive and negative control over sugar intake in mice.

Although the consumption of carbohydrates is needed for survival, their potent reinforcing properties drive obesity worldwide. In turn, sugar overconsumption reveals a major role for brain reward systems in regulating sugar intake. However, it remains elusive how different cell types within the reward circuitries control the initiation and termination of sugary meals. Here, we identified the distinct nucleus accumbens cell types that mediate the chemosensory versus postprandial properties of sweet sugars. Specifically, D1 neurons enhance sugar intake via specialized connections to taste ganglia, whereas D2 neurons mediate the termination of sugary meals via anatomical connections to circuits involved in appetite suppression. Consistently, D2, but not D1, neurons partially mediate the satiating effects of glucagon-like peptide 1 (GLP-1) agonists. Thus, these nucleus accumbens cell types function as a behavioral switch, enabling positive versus negative control over sugar intake. Our study contributes to unveiling the cellular and circuit substrates of sugar overconsumption.

Bacteriophage PRD1 as a nanoscaffold for drug loading.

Viruses are very attractive biomaterials owing to their capability as nanocarriers of genetic material. Efforts have been made to functionalize self-assembling viral protein capsids on their exterior or interior to selectively take up different payloads. PRD1 is a double-stranded DNA bacteriophage comprising an icosahedral protein outer capsid and an inner lipidic vesicle. Here, we report the three-dimensional structure of PRD1 in complex with the antipsychotic drug chlorpromazine (CPZ) by cryo-electron microscopy. We show that the jellyrolls of the viral major capsid protein P3, protruding outwards from the capsid shell, serve as scaffolds for loading heterocyclic CPZ molecules. Additional X-ray studies and molecular dynamics simulations show the binding modes and organization of CPZ molecules when complexed with P3 only and onto the virion surface. Collectively, we provide a proof of concept for the possible use of the lattice-like organisation and the quasi-symmetric morphology of virus capsomers for loading heterocyclic drugs with defined properties.

Structural basis for assembly of vertical single ß-barrel viruses.

The vertical double ß-barrel major capsid protein (MCP) fold, fingerprint of the PRD1-adeno viral lineage, is widespread in many viruses infecting organisms across the three domains of life. The discovery of PRD1-like viruses with two MCPs challenged the known assembly principles. Here, we present the cryo-electron microscopy (cryo-EM) structures of the archaeal, halophilic, internal membrane-containing Haloarcula californiae icosahedral virus 1 (HCIV-1) and Haloarcula hispanica icosahedral virus 2 (HHIV-2) at 3.7 and 3.8 Å resolution, respectively. Our structures reveal proteins located beneath the morphologically distinct two- and three-tower capsomers and homopentameric membrane proteins at the vertices that orchestrate the positioning of pre-formed vertical single ß-barrel MCP heterodimers. The cryo-EM based structures together with the proteomics data provide insights into the assembly mechanism of this type of viruses and into those with membrane-less double ß-barrel MCPs.

A Neural Circuit for Gut-Induced Reward.

The gut is now recognized as a major regulator of motivational and emotional states. However, the relevant gut-brain neuronal circuitry remains unknown. We show that optical activation of gut-innervating vagal sensory neurons recapitulates the hallmark effects of stimulating brain reward neurons. Specifically, right, but not left, vagal sensory ganglion activation sustained self-stimulation behavior, conditioned both flavor and place preferences, and induced dopamine release from Substantia nigra. Cell-specific transneuronal tracing revealed asymmetric ascending pathways of vagal origin throughout the CNS. In particular, transneuronal labeling identified the glutamatergic neurons of the dorsolateral parabrachial region as the obligatory relay linking the right vagal sensory ganglion to dopamine cells in Substantia nigra. Consistently, optical activation of parabrachio-nigral projections replicated the rewarding effects of right vagus excitation. Our findings establish the vagal gut-to-brain axis as an integral component of the neuronal reward pathway. They also suggest novel vagal stimulation approaches to affective disorders.

Is the SenseWear Armband accurate enough to quantify and estimate energy expenditure in healthy adults?

BACKGROUND: The SenseWear Armband (SWA) is a monitor that can be used to estimate energy expenditure (EE); however, it has not been validated in healthy adults. The objective of this paper was to study the validity of the SWA for quantifying EE levels. METHODS: Twenty-three healthy adults (age 40-55 years, mean: 48±3.42 years) performed different types of standardized physical activity (PA) for 10 minutes (rest, walking at 3 and 5 km·h-1, running at 7 and 9 km·h-1, and sitting/standing at a rate of 30 cycle·min-1). Participants wore the SWA on their right arm, and their EE was measured by indirect calorimetry (IC) the gold standard. RESULTS: There were significant differences between the SWA and IC, except in the group that ran at 9 km·h-1 (>9 METs). Bland-Altman analysis showed a BIAS of 1.56 METs (±1.83 METs) and limits of agreement (LOA) at 95% of -2.03 to 5.16 METs. There were indications of heteroscedasticity (R2 =0.03; P<0.05). Analysis of the receiver operating characteristic (ROC) curves showed that the SWA seems to be not sensitive enough to estimate the level of EE at highest intensities. CONCLUSIONS: The SWA is not as precise in estimating EE as IC, but it could be a useful tool to determine levels of EE at low intensities.

Integrated Control of Predatory Hunting by the Central Nucleus of the Amygdala.

Superior predatory skills led to the evolutionary triumph of jawed vertebrates. However, the mechanisms by which the vertebrate brain controls predation remain largely unknown. Here, we reveal a critical role for the central nucleus of the amygdala in predatory hunting. Both optogenetic and chemogenetic stimulation of central amygdala of mice elicited predatory-like attacks upon both insect and artificial prey. Coordinated control of cervical and mandibular musculatures, which is necessary for accurately positioning lethal bites on prey, was mediated by a central amygdala projection to the reticular formation in the brainstem. In contrast, prey pursuit was mediated by projections to the midbrain periaqueductal gray matter. Targeted lesions to these two pathways separately disrupted biting attacks upon prey versus the initiation of prey pursuit. Our findings delineate a neural network that integrates distinct behavioral modules and suggest that central amygdala neurons instruct predatory hunting across jawed vertebrates.

Membrane-assisted viral DNA ejection.

Genome packaging and delivery are fundamental steps in the replication cycle of all viruses. Icosahedral viruses with linear double-stranded DNA (dsDNA) usually package their genome into a preformed, rigid procapsid using the power generated by a virus-encoded packaging ATPase. The pressure and stored energy due to this confinement of DNA at a high density is assumed to drive the initial stages of genome ejection. Membrane-containing icosahedral viruses, such as bacteriophage PRD1, present an additional architectural complexity by enclosing their genome within an internal membrane vesicle. Upon adsorption to a host cell, the PRD1 membrane remodels into a proteo-lipidic tube that provides a conduit for passage of the ejected linear dsDNA through the cell envelope. Based on volume analyses of PRD1 membrane vesicles captured by cryo-electron tomography and modeling of the elastic properties of the vesicle, we propose that the internal membrane makes a crucial and active contribution during infection by maintaining the driving force for DNA ejection and countering the internal turgor pressure of the host. These novel functions extend the role of the PRD1 viral membrane beyond tube formation or the mere physical confinement of the genome. The presence and assistance of an internal membrane might constitute a biological advantage that extends also to other viruses that package their linear dsDNA to high density within an internal vesicle.

Separate circuitries encode the hedonic and nutritional values of sugar.

Sugar exerts its potent reinforcing effects via both gustatory and post-ingestive pathways. It is, however, unknown whether sweetness and nutritional signals engage segregated brain networks to motivate ingestion. We found in mice that separate basal ganglia circuitries mediated the hedonic and nutritional actions of sugar. During sugar intake, suppressing hedonic value inhibited dopamine release in ventral, but not dorsal, striatum, whereas suppressing nutritional value inhibited dopamine release in dorsal, but not ventral, striatum. Consistently, cell-specific ablation of dopamine-excitable cells in dorsal, but not ventral, striatum inhibited sugar's ability to drive the ingestion of unpalatable solutions. Conversely, optogenetic stimulation of dopamine-excitable cells in dorsal, but not ventral, striatum substituted for sugar in its ability to drive the ingestion of unpalatable solutions. Our data indicate that sugar recruits a distributed dopamine-excitable striatal circuitry that acts to prioritize energy-seeking over taste quality.

Speed and accuracy of taste identification and palatability: impact of learning, reward expectancy, and consummatory licking.

Despite decades of study, it remains a matter of controversy as to whether in rats taste identification is a rapid process that occurs in about 250-600 ms (one to three licks) or a slow process that evolves over seconds. To address this issue, we trained rats to perform a taste-cued two-response discrimination task (2-RDT). It was found that, after learning, regardless of intensity, the delivery of 10 µl of a tastant (e.g., NaCl or monopotassium glutamate, MPG) was sufficient to identify its taste with maximal accuracy within 400 ms. However, despite overtraining, rats rarely stopped licking in one lick. Thus, a one-drop lick reaction task was developed in which subjects had to rapidly stop licking after release of a stop signal (tastants including water) to obtain rewards. The faster they stopped licking, the greater the reward. Rats did not stop licking after receiving either hedonically positive or negative stop signals, and thus failed to maximize rewards even when reinforced with even larger rewards. In fact, the higher the sucrose concentration given as a stop signal, the greater the number of consummatory licks elicited. However, with a stop signal of 2 mM quinine HCl, they stopped licking in ~370 ms, a time faster than that for sucrose or water, thus showing that in this rapid period, quinine HCl evoked an unpalatable response. Indeed, only when rats licked an empty sipper tube would they usually elicit a single lick to obtain a reward (operant licking). In summary, these data indicate that within 400 ms, taste identification and palatability, must either occur simultaneously or with marked overlap.

PAU-SA: a synthetic aperture interferometric radiometer test bed for potential improvements in future missions.

The Soil Moisture and Ocean Salinity (SMOS) mission is an Earth Explorer Opportunity mission from the European Space Agency (ESA). Its goal is to produce global maps of soil moisture and ocean salinity using the Microwave Imaging Radiometer by Aperture Synthesis (MIRAS). The purpose of the Passive Advanced Unit Synthetic Aperture (PAU-SA) instrument is to study and test some potential improvements that could eventually be implemented in future missions using interferometric radiometers such as the Geoestacionary Atmosferic Sounder (GAS), the Precipitation and All-weather Temperature and Humidity (PATH) and the Geostationary Interferometric Microwave Sounder (GIMS). Both MIRAS and PAU-SA are Y-shaped arrays with uniformly distributed antennas, but the receiver topology and the processing unit are quite different. The purpose of this work is to identify the elements in the MIRAS's design susceptible of improvement and apply them in the PAU-SA instrument demonstrator, to test them in view of these future interferometric radiometer missions.

Transitions between sleep and feeding states in rat ventral striatum neurons.

Neurons in the nucleus accumbens (NAc) have been shown to participate in several behavioral states, including feeding and sleep. However, it is not known if the same neuron participates in both states and, if so, how similar are the responses. In addition, since the NAc contains several cell types, it is not known if each type participates in the transitions associated with feeding and sleep. Such knowledge is important for understanding the interaction between two different neural networks. For these reasons we recorded ensembles of NAc neurons while individual rats volitionally transitioned between the following states: awake and goal directed, feeding, quiet-awake, and sleeping. We found that during both feeding and sleep states, the same neurons could increase their activity (be activated) or decrease their activity (be inactivated) by feeding and/or during sleep, thus indicating that the vast majority of NAc neurons integrate sleep and feeding signals arising from spatially distinct neural networks. In contrast, a smaller population was modulated by only one of the states. For the majority of neurons in either state, we found that when one population was excited, the other was inhibited, suggesting that they act as a local circuit. Classification of neurons into putative interneurons [fast-spiking interneurons (pFSI) and choline acetyltransferase interneurons (pChAT)] and projection medium spiny neurons (pMSN) showed that all three types are modulated by transitions to and from feeding and sleep states. These results show, for the first time, that in the NAc, those putative inhibitory interneurons respond similarly to pMSN projection neurons and demonstrate interactions between NAc networks involved in sleep and feeding.

A general analysis of the impact of digitization in microwave correlation radiometers.

This study provides a general framework to analyze the effects on correlation radiometers of a generic quantization scheme and sampling process. It reviews, unifies and expands several previous works that focused on these effects separately. In addition, it provides a general theoretical background that allows analyzing any digitization scheme including any number of quantization levels, irregular quantization steps, gain compression, clipping, jitter and skew effects of the sampling period.

Physical activity among Spanish adolescents: relationship with their relatives' physical activity - the AVENA study.

Identifying factors related to physical activity levels in young people is important for a more efficient health promotion. The aims of this study were to assess physical activity levels in a national sample of urban Spanish adolescents, and to examine the association between significant others' physical activity (father, mother, brother, sister, and close friends) and that of the adolescents. The present study comprised 2260 adolescents (1157 boys, 1103 girls) aged 13.0-18.5 years participating in the AVENA Study. Both the adolescents' physical activity and that of their relatives and close friends was assessed by questionnaire. The odds of being active were higher in boys than girls (odds ratio = 2.79, 95% confidence interval = 2.34-3.33) and tended to decrease across age groups in both boys and girls. Father's and older brother's physical activity was associated with boys' physical activity, while that of any significant other was associated with girls' physical activity. When both parents reported being active, boys had nearly two times higher odds of being active and girls had nearly three times higher odds of being active. The physical activity levels of Spanish adolescents are in line with those previously reported. Physical activity levels in girls are strongly related to the physical activity of any significant other, whereas physical activity levels in boys are only related to their male relatives' physical activity.

Socio-economic factors and active commuting to school in urban Spanish adolescents: the AVENA study.

BACKGROUND: This study aimed: (i) to describe the patterns of commuting to school in urban Spanish adolescents; and (ii) to examine the associations between active commuting to school (ACS) and socio-economic factors. METHODS: From the AVENA Study, 2183 adolescents (1142 females) aged 13-18.5 years were gathered. Mode and time of transportation to school were self-reported by the adolescents. Parental education level (primary, secondary or university degree), parental professional level (managerial, skilled worker or unskilled worker/unemployed) and the type of school (public or private) were self-reported by the parents. The relationships between ACS and socio-economic factors were analysed by binary logistic regression. RESULTS: Nearly <65% of the adolescents reported ACS and 83% of them spent <15 min travelling to school. In male adolescents, maternal primary education level showed an odds ratio (OR) of 1.55 (95% confidence interval, 1.12-2.15), with respect to mothers with a university degree. In female adolescents, mothers with a primary education level showed an OR of 0.68 (0.50-0.92), with respect to mothers with a university degree. Low maternal professional level showed an OR of 1.70 (1.29-2.24), with respect to high maternal professional levels. Students attending public schools showed an OR of 3.47 (2.46-4.90), with respect to students from private schools. CONCLUSION: Most of the adolescents actively commuted to school, yet the time spent commuting was low. Socio-economic level seems to be inversely related to the ACS in adolescents. Type of school and maternal educational level were the main predictors of ACS.

New passive instruments developed for ocean monitoring at the remote sensing lab-universitat politècnica de catalunya.

Lack of frequent and global observations from space is currently a limiting factor in many Earth Observation (EO) missions. Two potential techniques that have been proposed nowadays are: (1) the use of satellite constellations, and (2) the use of Global Navigation Satellite Signals (GNSS) as signals of opportunity (no transmitter required). Reflectometry using GNSS opportunity signals (GNSS-R) was originally proposed in 1993 by Martin-Neira (ESA-ESTEC) for altimetry applications, but later its use for wind speed determination has been proposed, and more recently to perform the sea state correction required in sea surface salinity retrievals by means of L-band microwave radiometry (T(B)). At present, two EO space-borne missions are currently planned to be launched in the near future: (1) ESA's SMOS mission, using a Y-shaped synthetic aperture radiometer, launch date November 2nd, 2009, and (2) NASA-CONAE AQUARIUS/SAC-D mission, using a three beam push-broom radiometer. In the SMOS mission, the multi-angle observation capabilities allow to simultaneously retrieve not only the surface salinity, but also the surface temperature and an "effective" wind speed that minimizes the differences between observations and models. In AQUARIUS, an L-band scatterometer measuring the radar backscatter (σ(0)) will be used to perform the necessary sea state corrections. However, none of these approaches are fully satisfactory, since the effective wind speed captures some sea surface roughness effects, at the expense of introducing another variable to be retrieved, and on the other hand the plots (T(B)-σ(0)) present a large scattering. In 2003, the Passive Advance Unit for ocean monitoring (PAU) project was proposed to the European Science Foundation in the frame of the EUropean Young Investigator Awards (EURYI) to test the feasibility of GNSS-R over the sea surface to make sea state measurements and perform the correction of the L-band brightness temperature. This paper: (1) provides an overview of the Physics of the L-band radiometric and GNSS reflectometric observations over the ocean, (2) describes the instrumentation that has been (is being) developed in the frame of the EURYI-funded PAU project, (3) the ground-based measurements carried out so far, and their interpretation in view of placing a GNSS-reflectometer as secondary payload in future SMOS follow-on missions.