Solar wind entry into the high-latitude terrestrial magnetosphere during geomagnetically quiet times.

An understanding of the transport of solar wind plasma into and throughout the terrestrial magnetosphere is crucial to space science and space weather. For non-active periods, there is little agreement on where and how plasma entry into the magnetosphere might occur. Moreover, behaviour in the high-latitude region behind the magnetospheric cusps, for example, the lobes, is poorly understood, partly because of lack of coverage by previous space missions. Here, using Cluster multi-spacecraft data, we report an unexpected discovery of regions of solar wind entry into the Earth's high-latitude magnetosphere tailward of the cusps. From statistical observational facts and simulation analysis we suggest that these regions are most likely produced by magnetic reconnection at the high-latitude magnetopause, although other processes, such as impulsive penetration, may not be ruled out entirely. We find that the degree of entry can be significant for solar wind transport into the magnetosphere during such quiet times.

Observations of ionospheric electron beams in the plasma sheet.

Electrons streaming along the magnetic field direction are frequently observed in the plasma sheet of Earth's geomagnetic tail. The impact of these field-aligned electrons on the dynamics of the geomagnetic tail is however not well understood. Here we report the first detection of field-aligned electrons with fluxes increasing at ~1 keV forming a "cool" beam just prior to the dissipation of energy in the current sheet. These field-aligned beams at ~15 R(E) in the plasma sheet are nearly identical to those commonly observed at auroral altitudes, suggesting the beams are auroral electrons accelerated upward by electric fields parallel (E([parallel])) to the geomagnetic field. The density of the beams relative to the ambient electron density is δn(b)/n(e)~5-13% and the current carried by the beams is ~10(-8)-10(-7) A m(-2). These beams in high ß plasmas with large density and temperature gradients appear to satisfy the Bohm criteria to initiate current driven instabilities.

Entropy generation across Earth's collisionless bow shock.

Earth's bow shock is a collisionless shock wave but entropy has never been directly measured across it. The plasma experiments on Cluster and Double Star measure 3D plasma distributions upstream and downstream of the bow shock allowing calculation of Boltzmann's entropy function H and his famous H theorem, dH/dt≤0. The collisionless Boltzmann (Vlasov) equation predicts that the total entropy does not change if the distribution function across the shock becomes nonthermal, but it allows changes in the entropy density. Here, we present the first direct measurements of entropy density changes across Earth's bow shock and show that the results generally support the model of the Vlasov analysis. These observations are a starting point for a more sophisticated analysis that includes 3D computer modeling of collisionless shocks with input from observed particles, waves, and turbulences.

The melanin-concentrating hormone (MCH) system in an animal model of depression-like behavior.

Selective breeding for divergence in locomotion in a novel environment (bHR, bred High-Responder; bLR, bred Low-Responder) correlates with stress-reactivity, spontaneous anxiety-like behaviors and predicts vulnerability in a rodent model of depression. Identifying genetic factors that may account for such vulnerability are key determinants not only for the illness outcome but also for the development of better-tailored treatment options. Melanin-concentrating hormone (MCH) is a neuropeptide that exhibits some of the hallmarks of a regulator of affective states. The aim of this study was to ascertain the role of the MCH system in depression-like behaviors in bHR vs. bLR rats. bLR rats showed a 44% increase in hypothalamic pMCH mRNA and a 14% decrease in hippocampal CA1 MCH1R mRNA when compared to bHR rats. Interestingly, the amount of time that rats spent immobile in the FST (depressive-like behavior) correlated positively with the amount of hypothalamic pMCH mRNA and negatively with that of hippocampal CA1 MCH1R. The results indicate that the bLR-bHR is a useful rat model to investigate individual basal genetic differences that participate in the monitoring of emotional responsiveness (i.e., depression- and anxiety-like behaviors). They also point to the MCH system (i.e., chronically higher pMCH expression and consequently receptor down-regulation) as a candidate biomarker for the severity of depressive-like behavior. The data indicate that MCH1R participates in the modulation of depression-like behavior through a process that involves the CA1 region of the hippocampus, supporting the possible use of MCH1R antagonists in the treatment of depression.

Nonlinear development of shocklike structure in the solar wind.

We report first in situ multispacecraft observations of nonlinear steepening of compressional pulses in the solar wind upstream of Earth's bow shock. The magnetic field of a compressional pulse formed at the upstream edge of density holes is shown to suddenly break and steepen into a shocklike structure. During the early phase of development thermalization of ions is insignificant. Substantial thermalization of ions occurs as gyrating ions are observed at the steepened edge. These observations indicate that the mechanisms causing the dissipation of magnetic fields (currents) and ions are different in the early phase of shock development.

Solitary electromagnetic pulses detected with super-Alfvénic flows in Earth's geomagnetic tail.

Solitary nonlinear (deltaB/B>>1) electromagnetic pulses have been detected in Earth's geomagnetic tail accompanying plasmas flowing at super-Alfvénic speeds. The pulses in the current sheet had durations of approximately 5 s, were left-hand circularly polarized, and had phase speeds of approximately the Alfvén speed in the plasma frame. These pulses were associated with a field-aligned current J(parallel) and observed in low density (approximately 0.3 cm(-3)), high temperature (T(e) approximately T(i) approximately 3x10(7) K), and beta approximately 10 plasma that included electron and ion beams streaming along B. The wave activity was enhanced from below the ion cyclotron frequency to electron cyclotron and upper hybrid frequencies. The detailed properties suggest the pulses are nonlinearly steepened ion cyclotron or Alfvén waves.

Design of reverse osmosis (RO) water treatment networks subject to fouling.

Identifying the optimal design for an RO membrane network is not straightforward when significant fouling occurs. The most robust optimal network design will feature the minimal capital and operating costs over the anticipated lifetime of the plant, and is therefore strongly dependent on the fouling behaviour and associated mitigation. This study considers the case where the likely fouling behaviour is known and investigates how to incorporate this knowledge into the design and operation of a network. The optimisation task is complicated by the highly non-linear nature of the problem owing to membrane behaviour, fouling behaviour, network interactions and operating parameter constraints (pressures and flows). In this work, fouling is modelled as an exponential decay in membrane permeability. Three optimisation approaches are used to evaluate candidate networks: (i) laborious comparison of pre-selected individual network designs; (ii) deterministic gradient search methods, and (iii) a simulated-annealing-based hybrid stochastic-deterministic method. All of the approaches consider various configurations of a two-stage network with a maximum of three membrane units in each stage, represented in a superstructure model. The results from the approaches are compared and the most effective method for network design is discussed.

National Retail Data Monitor for public health surveillance.

The National Retail Data Monitor (NRDM) is a public health surveillance tool that collects and analyzes daily sales data for over-the-counter (OTC) health-care products. NRDM collects sales data for selected OTC health-care products in near real time from >15,000 retail stores and makes them available to public health officials. NRDM is one of the first examples of a national data utility for public health surveillance that collects, redistributes, and analyzes daily sales-volume data of selected health-care products, thereby reducing the effort for both data providers and health departments.

The role of religion in predicting adolescent alcohol use and problem drinking.

OBJECTIVE: There are racial differences in adolescents' propensity to consume alcohol--with white adolescents tending to consume more alcohol than black adolescents--but there is no clear explanation for why such differences exist. The purpose of this study was to examine the relationship between religiosity, a cultural factor that is not well understood currently, and racial differences in adolescent alcohol use. METHOD: Participants were white and black ninth-grade adolescents (N = 899; 54% female, 57.5% white) involved in a 3-year longitudinal study of ways to reduce alcohol use and sexual risk-taking behavior among adolescents in Ohio and Kentucky. RESULTS: Findings indicate that religiosity is differentially associated with alcohol use and problem drinking for white and black adolescents. Religious service attendance was the most significant predictor of alcohol use for black adolescents, whereas religious fundamentalism was most important for white adolescents. In contrast, frequency of prayer was the significant predictor of problem drinking for black adolescents, whereas the level of importance placed on religion was the significant predictor for white adolescents. Important gender differences also emerged in both prediction models and are discussed. CONCLUSIONS: Since there is great heterogeneity among adolescents (in terms of race and gender) in their alcohol use and misuse, the "one-size-fits-all" approach to alcohol treatment and prevention is likely inappropriate. Moreover, conceptualizations of alcohol use and misuse, and its prevention and treatment, should include the consideration of such key cultural factors as religiosity.

A novel CD8-independent high-avidity cytotoxic T-lymphocyte response directed against an epitope in the phosphoprotein of the paramyxovirus simian virus 5.

Adoptive transfer studies have shown that cytotoxic T lymphocytes (CTL) of high avidity, capable of recognizing low levels of peptide-MHC I molecules, are more efficient at reducing viral titers than are low-avidity CTL, thus establishing CTL avidity as a critical parameter for the ability of a CTL to clear virus in vivo. It has been well documented that CTL of high avidity are relatively CD8 independent, whereas low-avidity CTL require CD8 engagement in order to become activated. In this study we have analyzed the antiviral CTL response elicited following infection with the paramyxovirus simian virus 5 (SV5). We have identified the immunodominant and subdominant CTL responses and subsequently assessed the avidity of these responses by their CD8 dependence. This is the first study in which the relationship between immunodominance and CTL avidity has been investigated. The immunodominant response was directed against an epitope present in the viral M protein, and subdominant responses were directed against epitopes present in the P, F, and HN proteins. Similarly to other CTL responses we have analyzed, the immunodominant response and the subdominant F and HN responses were comprised of both high- and low-avidity CTL. However, the subdominant response directed against the epitope present in the P protein is novel, as it is exclusively high avidity. This high-avidity response is independent of both the route of infection and expression by recombinant SV5. A further understanding of the inherent properties of P that elicit only high-avidity CTL may allow for the design of more efficacious vaccine vectors that preferentially elicit high-avidity CTL in vivo.

Modulation of outward potassium currents in aligned cultures of neonatal rat ventricular myocytes during phorbol ester-induced hypertrophy.

Protein kinase C-stimulating phorbol esters induce a strong hypertrophic response when applied in vitro to cardiac ventricular myocytes. The aim of this study was to determine if this in vitro model of hypertrophy is associated with changes in the expression of voltage-gated K(+)channels. Myocytes were isolated from 3--4-day-old neonatal rats and cultured on aligned collagen thin gels. Membrane currents were measured with the use of the whole-cell arrangement of the patch clamp technique and the expression levels of the Kv1.4, Kv4.2 and Kv2.1 alpha subunits quantified using Western blot analysis. Voltage steps positive to -30 mV resulted in the activation of both a transient (I(to)) and a sustained (I(sus)) component of outward K(+)current in the aligned myocytes. Overnight exposure to phorbol 12-myristate 13-acetate (PMA) caused a 55% increase in myocyte size and a three-fold reduction in the peak amplitude of I(to). No differences in the half-maximal voltages required for activation and steady-state inactivation were observed between I(to)measured in control and PMA-treated myocytes. In contrast, PMA treatment resulted in a 62% increase in a tetraethylammonium-sensitive component of I(sus)(TEA-I(sus)) and was associated with the appearance of a slow component of current decay. Expression levels of the Kv1.4 and Kv4.2 alpha subunits were strongly depressed in the hypertrophic myocytes, while the density of the Kv2.1 alpha subunit was enhanced. PMA-induced changes in the Kv alpha subunits were partially prevented through inhibition of the mitogen-activated protein kinase (MAPK) pathway. Thus, PMA-induced hypertrophy of cultured ventricular myocytes is associated with an altered expression of voltage-gated K(+)channels.

The V protein of human parainfluenza virus 2 antagonizes type I interferon responses by destabilizing signal transducer and activator of transcription 2.

Type I interferon (IFN) induces antiviral responses through the activation of the ISGF3 transcription factor complex that contains the subunit proteins STAT1, STAT2, and p48/ISGF3 gamma/IRF9. The ability of some human paramyxoviruses to overcome IFN actions by specific proteolysis of STAT proteins has been examined. Infection of cells with type 2, but not type 1 or type 3 human parainfluenza virus (HPIV) leads to a loss of cellular STAT2 protein. Expression of a single HPIV2 protein derived from the V open reading frame blocks IFN-dependent transcriptional responses in the absence of other viral proteins. The loss of IFN response is due to V-protein-induced proteolytic degradation of STAT2. Expression of HPIV2 V causes the normally stable STAT2 protein to be rapidly degraded, and this proteolytic activity can be partially alleviated by proteasome inhibition. No V-protein-specific effects on STAT2 mRNA levels were observed. The results indicate that the V protein of HPIV2 is sufficient to recognize and target a specific cellular transcription factor for destruction by cellular machinery.

Integrating harm reduction therapy and traditional substance abuse treatment.

One-size-fits-all therapy has not worked well for a majority of substance users seeking help. New approaches to substance abuse treatment are desperately needed. Traditional models of service delivery offer little, if any, help to people who may not choose abstinence as a goal. To address this concern, the Bridging the Gap Conference was sponsored by the San Francisco Department of Public Health. The overall goals of the conference were to improve standards of care, develop best practice principles for integrating harm reduction approaches into traditional substance abuse services, and increase the accessibility of quality services to people in need of alcohol and drug treatment. G. Alan Marlatt gave a keynote address on the integration of harm reduction therapy into traditional treatment services, an expanded version of which is presented in this article. Such integration would broaden the scope of services available to a larger group of consumers of substance abuse treatment. Furthermore, harm reduction therapy would infuse traditional treatment practices with scientifically-based pragmatism that pays close attention to individual and community public health needs. Because of its tolerance of treatment goals other than abstinence, harm reduction therapy offers the greatest hope to expand the availability of substance abuse services to people who have not benefited from traditional abstinence-based treatment models.

RNA replication from the simian virus 5 antigenomic promoter requires three sequence-dependent elements separated by sequence-independent spacer regions.

We have previously shown for the paramyxovirus simian virus 5 (SV5) that a functional promoter for RNA replication requires proper spacing between two discontinuous elements: a 19-base segment at the 3' terminus (conserved region I [CRI]) and an 18-base internal region (CRII) that is contained within the coding region of the L protein gene. In the work described here, we have used a reverse-genetics system to determine if the 53-base segment between CRI and CRII contains additional sequence-specific signals required for optimal replication or if this segment functions solely as a sequence-independent spacer region. A series of copyback defective interfering minigenome analogs were constructed to contain substitutions of nonviral sequences in place of bases 21 to 72 of the antigenomic promoter, and the relative level of RNA replication was measured by Northern blot analysis. The results from our mutational analysis indicate that in addition to CRI and CRII, optimal replication from the SV5 antigenomic promoter requires a third sequence-dependent element located 51 to 66 bases from the 3' end of the RNA. Minigenome RNA replication was not affected by changes in the either the position of this element in relation to CRI and CRII or the predicted hexamer phase of NP encapsidation. Thus, optimal RNA replication from the SV5 antigenomic promoter requires three sequence-dependent elements, CRI, CRII and bases 51 to 66.

Increased readthrough transcription across the simian virus 5 M-F gene junction leads to growth defects and a global inhibition of viral mRNA synthesis.

Recombinant simian virus 5 (rSV5) mutants containing substitutions in the M-F intergenic region were generated to determine the effect of increased readthrough transcription on the paramyxovirus growth cycle. We have previously shown, using an SV5 dicistronic minigenome, that replacement of the 22-base M-F intergenic region with a foreign sequence results in a template (Rep22) that directs very high levels of M-F readthrough transcription. An rSV5 containing the Rep22 substitution grew slower and to final titers that were 50- to 80-fold lower than those of wild-type (WT) rSV5. Cells infected with the Rep22 virus produced very low levels of monocistronic M and F mRNA, consistent with the M-F readthrough phenotype. Surprisingly, Rep22 virus-infected cells also displayed a global decrease in the accumulation of viral mRNA from genes located upstream and downstream of the M-F junction, and overall viral protein synthesis was reduced. Second-site revertants of the Rep22 virus that had regained WT transcription and growth properties contained a single base substitution that increased the M gene end U tract from four to eight residues, suggesting that the growth defects originated from higher-than-normal M-F readthrough transcription. Thus, the primary growth defect for the Rep22 virus appears to be in viral RNA synthesis and not in morphogenesis. A second rSV5 virus (G14), which contained a different foreign M-F intergenic sequence, grew to similar or slightly higher titers than WT rSV5 in some cell types and produced ~1.5- to 2-fold more mRNA and viral protein. The data support the hypothesis that inhibition of Rep22 virus growth is due to increased access by the polymerase to the 5' end of the genome and to the resulting overexpression of L protein. We propose that the elevated naturally occurring M-F readthrough which is characteristic of many paramyxoviruses serves as a mechanism to fine-tune the level of polymerase that is optimal for virus growth.

Spacing constraints on reinitiation of paramyxovirus transcription: the gene end U tract acts as a spacer to separate gene end from gene start sites.

The paramyxovirus gene end U tracts are thought to serve as templates for the addition of a 3' polyA tail to viral mRNAs. The goal of the work described here was to determine the function in transcription of the naturally occurring variability in length of the gene end U tracts of the paramyxovirus simian virus 5 (SV5). An anchored RT-PCR assay was developed to test the hypothesis that the variable U tracts template the addition of variable lengths of polyA tails to mRNAs. The results showed that although the SV5 NP, M, and SH genes encode U tracts of seven, four, and six U residues, respectively, their mRNAs contain similar polyA tails of approximately 250-290 bases. These results indicate that the variable gene end U tracts are functionally equivalent in directing polyadenylation. A reverse genetics system based on a dicistronic minigenome containing the SH-HN gene junction was used to test the hypothesis that the variable U tracks affect the efficiency of transcription termination. Minigenome templates containing an SH gene end with a long U tract of six residues (U6) directed efficient transcription termination and reinitiation at the downstream HN start site with no nucleotide preference for the downstream intergenic region. Surprisingly, truncating the SH gene end U tract to four residues (U4) did not affect SH termination but, rather, reduced downstream HN reinitiation to 20-30% of wild-type levels. Efficient HN reinitiation could be restored to mutant U4 templates in either of two ways: by increasing the U-tract length from four to six residues or by increasing the length of the intergenic region. Efficient HN reinitiation required a minimum of six bases between the last nucleotide in SH and the first nucleotide in HN. We propose that for some paramyxoviruses, the gene end U tract serves a previously unrecognized role as a spacer region between the gene end and gene start sites.

Highly diverse intergenic regions of the paramyxovirus simian virus 5 cooperate with the gene end U tract in viral transcription termination and can influence reinitiation at a downstream gene.

A dicistronic minigenome containing the M-F gene junction was used to determine the role of the simian virus 5 (SV5) intergenic regions in transcription. The M-F junction differs from the other SV5 junctions by having a short M gene end U tract of only four residues (U4 tract) and a 22-base M-F intergenic sequence between the M gene end and F gene start site. Replacing the 22-base M-F intergenic region with nonviral sequences resulted in a minigenome template (Rep 22) that was defective in termination at the end of the M gene. Efficient M gene termination could be restored to the mutant Rep 22 template in either of two ways: by increasing the U tract length from four to six residues or by restoring a G residue immediately downstream of the wild-type (WT) U4 tract. In a dicistronic SH-HN minigenome, a U4-G combination was functionally equivalent to the naturally occurring SH U6-A gene end in directing SH transcription termination. In addition to affecting termination, the M-F intergenic region also influenced polymerase reinitiation. In the context of the WT U4-G M gene end, substituting nonviral sequences into the M-F intergenic region had a differential effect on F gene reinitiation, where some but not all nonviral sequences inhibited reinitiation. The inhibition of F gene reinitiation correlated with foreign sequences having a high C content. Deleting 6 bases or inserting 18 additional nucleotides into the middle of the 22-base M-F intergenic segment did not influence M gene termination or F gene reinitiation, indicating that M-F intergenic length per se is not a important factor modulating the SV5 polymerase activity. Our results suggest that the sequence diversity at an SV5 gene junction reflects specific combinations which may differentially affect SV5 gene expression and provide an additional level of transcriptional control beyond that which results from the distance of a gene from the 3' end promoter.

RNA replication for the paramyxovirus simian virus 5 requires an internal repeated (CGNNNN) sequence motif.

A functional RNA replication promoter for the paramyxovirus simian virus 5 (SV5) requires two essential and discontinuous elements: 19 bases at the 3' terminus (conserved region I) and an 18-base internal region (conserved region II [CRII]) that is contained within the coding region of the L protein gene. A reverse-genetics system was used to determine the sequence requirements for the internal CRII element to function in RNA replication. A series of copyback defective interfering (DI) RNA analogs were constructed to contain point mutations in the 18 nucleotides composing CRII, and their relative replication levels were analyzed. The results indicated that SV5 DI RNA replication was reduced by substitutions for two CG dinucleotides, which in the nucleocapsid template are in the first two positions of the first two hexamers of CRII nucleotides. Substitutions for other bases within CRII did not reduce RNA synthesis. Thus, two consecutive 5'-CGNNNN-3' hexamers form an important sequence in the SV5 CRII promoter element. The position of the CG dinucleotide within the SV5 leader and antitrailer promoters was highly conserved among other members of the Rubulavirus genus, but this motif differed significantly in both sequence and position from that previously identified for Sendai virus. The possible roles of the CRII internal promoter element in paramyxovirus RNA replication are discussed.