An Intermediate State of the {gamma}-Aminobutyric Acid Transporter GAT1 Revealed by Simultaneous Voltage Clamp and Fluorescence

The rat {gamma}-aminobutyric acid transporter GAT1 expressed in Xenopus oocytes was labeled at Cys74, and at one or more other sites, by tetramethylrhodamine-5-maleimide, without significantly altering GAT1 function. Voltage-jump relaxation analysis showed that fluorescence increased slightly and monotonically with hyperpolarization; the fluorescence at -140 mV was ~0.8% greater than at +60 mV. The time course of the fluorescence relaxations was mostly described by a single exponential with voltage-dependent but history-independent time constants ranging from ~20 ms at +60 mV to ~150 ms at -140 mV. The fluorescence did not saturate at the most negative potentials tested, and the midpoint of the fluorescence–voltage relation was at least 50 mV more negative than the midpoint of the charge–voltage relation previously identified with Na+ binding to GAT1. The presence of {gamma}-aminobutyric acid did not noticeably affect the fluorescence waveforms. The fluorescence signal depended on Na+ concentration with a Hill coefficient approaching 2. Increasing Cl- concentration modestly increased and accelerated the fluorescence relaxations for hyperpolarizing jumps. The fluorescence change was blocked by the GAT1 inhibitor, NO-711. For the W68L mutant of GAT1, the fluorescence relaxations occurred only during jumps to high positive potentials, in agreement with previous suggestions that this mutant is trapped in one conformational state except at these potentials. These observations suggest that the fluorescence signals monitor a novel state of GAT1, intermediate between the E*out and Eout states of Hilgemann, D.W., and C.-C. Lu (1999. J. Gen. Physiol. 114:459–476). Therefore, the study provides verification that conformational changes occur during GAT1 function

Perceived barriers to pediatrician and family practitioner participation in pediatric clinical trials: Findings from the Clinical Trials Transformation Initiative.

Despite legislation to stimulate pediatric drug development through clinical trials, enrolling children in trials continues to be challenging. Non-investigator (those who have never served as a clinical trial investigator) providers are essential to recruitment of pediatric patients, but little is known regarding the specific barriers that limit pediatric providers from participating in and referring their patients to clinical trials. We conducted an online survey of pediatric providers from a wide variety of practice types across the United States to evaluate their attitudes and awareness of pediatric clinical trials. Using a 4-point Likert scale, providers described their perception of potential barriers to their practice serving as a site for pediatric clinical trials. Of the 136 providers surveyed, 52/136 (38%) had previously referred a pediatric patient to a trial, and only 17/136 (12%) had ever been an investigator for a pediatric trial. Lack of awareness of existing pediatric trials was a major barrier to patient referral by providers, in addition to consideration of trial risks, distance to the site, and time needed to discuss trial participation with parents. Overall, providers perceived greater challenges related to parental concerns and parent or child logistical barriers than study implementation and ethics or regulatory barriers as barriers to their practice serving as a trial site. Providers who had previously been an investigator for a pediatric trial were less likely to be concerned with potential barriers than non-investigators. Understanding the barriers that limit pediatric providers from collaboration or inhibit their participation is key to designing effective interventions to optimize pediatric trial participation

Outcome Assessment using Connected Vehicle Data to Justify Signal Investments to Decision Makers

Elected officials and decision makers are increasingly seeking outcome assessment of capital projects, such as corridor signal modernization and adaptive control projects. This paper describes the use of connected vehicle data to perform corridor travel time outcome assessment along five corridors in the greater Philadelphia, Pennsylvania area. These corridors are comprised of a total of 2,184 signals and are considered five of the most critical corridors in this region, experiencing a high volume of traffic, with AADT greater than 30,000 vehicles. These corridors were evaluated for six weeks before and after the adaptive installation through the use of private-sector segment speed data. Medians and interquartile ranges of travel times were used to assess the impact on arterial progression. Various graphs, charts, and figures produced through web tools and traditional metrics provide a user-interactive component to the dashboards. In addition, user cost reductions and CO2 emission impacts were also determined. Four out of the five corridors had substantial reductions of arterial travel times that amounted to approximately $36.6 million in annualized user benefits

Sparse geometric graphs with small dilation

Given a set S of n points in R^D, and an integer k such that 0 <= k < n, we show that a geometric graph with vertex set S, at most n - 1 + k edges, maximum degree five, and dilation O(n / (k+1)) can be computed in time O(n log n). For any k, we also construct planar n-point sets for which any geometric graph with n-1+k edges has dilation Omega(n/(k+1)); a slightly weaker statement holds if the points of S are required to be in convex position

Low-temperature thermochronometry along the Kunlun and Haiyuan Faults, NE Tibetan Plateau: Evidence for kinematic change during late-stage orogenesis

The Tibetan Plateau is a prime example of a collisional orogen with widespread strike-slip faults whose age and tectonic significance remain controversial. We present new low-temperature thermochronometry to date periods of exhumation associated with Kunlun and Haiyuan faulting, two major strike-slip faults within the northeastern margin of Tibet. Apatite and zircon (U-Th)/He and apatite fission-track ages, which record exhumation from ~2 to 6 km crustal depths, provide minimum bounds on fault timing. Results from Kunlun samples show increased exhumation rates along the western fault segment at circa 12–8 Ma with a possible earlier phase of motion from ~30–20 Ma, along the central fault segment at circa 20–15 Ma, and along the eastern fault segment at circa 8–5 Ma. Combined with previous studies, our results suggest that motion along the Haiyuan fault may have occurred as early as ~15 Ma along the western/central fault segment before initiating at least by 10–8 Ma along the eastern fault tip. We relate an ~250 km wide zone of transpressional shear to synchronous Kunlun and Haiyuan fault motion and suggest that the present-day configuration of active faults along the northeastern margin of Tibet was likely established since middle Miocene time. We interpret the onset of transpression to relate to the progressive confinement of Tibet against rigid crustal blocks to the north and expansion of crustal thickening to the east during the later stages of orogen development

Meningococcal Disease in Patients With Human Immunodeficiency Virus Infection: A Review of Cases Reported Through Active Surveillance in the United States, 2000-2008.

BackgroundAlthough human immunodeficiency virus (HIV) infection is an established risk factor for several bacterial infections, the association between HIV infection and meningococcal disease remains unclear.MethodsExpanded chart reviews were completed on persons with meningococcal disease and HIV infection reported from 2000 through 2008 from 9 US sites participating in an active population-based surveillance system for meningococcal disease. The incidence of meningococcal disease among patients meeting Centers for Disease Control and Prevention acquired immune deficiency syndrome (AIDS) surveillance criteria was estimated using data from the National HIV Surveillance System for the participating sites.ResultsThirty-three cases of meningococcal disease in individuals with HIV infection were reported from participating sites, representing 2.0% of all reported meningococcal disease cases. Most (75.8%) persons with HIV infection were adult males aged 25 to 64 years old. Among all meningococcal disease cases aged 25 to 64 years old, case fatality ratios were similar among HIV-infected and HIV-uninfected persons (13.3% vs 10.6%; P = .6). The cumulative, mean incidence of meningococcal disease among patients aged 25 to 64 years old with HIV infection ever classified as AIDS was 3.5 cases per 100000 person years (95% confidence interval [CI], 2.1-5.6), compared with 0.3 cases per 100000 person years (95% CI, 0.3-0.3) for persons of the same age group not reported to have AIDS (relative risk = 12.9; 95% CI, 7.9-20.9).ConclusionsIndividuals with HIV infection meeting the AIDS surveillance case definition have a higher incidence of meningococcal disease compared with the general adult population

Search for electric dipole moments at storage rings

Permanent electric dipole moments (EDMs) violate parity and time reversal symmetry. Within the Standard Model (SM) they are many orders of magnitude below present experimental sensitivity. Many extensions of the SM predict much larger EDMs, which are therefore an excellent probe for the existence of "new physics". Until recently it was believed that only electrically neutral systems could be used for sensitive searches of EDMs. With the introduction of a novel experimental method, high precision for charged systems will be within reach as well. The features of this method and its possibilities are discussed.Comment: Proc. EXA2011, 6 pages; http://www.springerlink.com/content/45l35376832vhrg0

Instrumentation Development for In Situ 40Ar/39Ar Planetary Geochronology

The chronology of the Solar System, particularly the timing of formation of extra-terrestrial bodies and their features, is an outstanding problem in planetary science. Although various chronological methods for in situ geochronology have been proposed (e.g., Rb-Sr, K-Ar), and even applied (K-Ar), the reliability, accuracy, and applicability of the 40Ar/39Ar method makes it by far the most desirable chronometer for dating extra-terrestrial bodies. The method however relies on the neutron irradiation of samples, and thus a neutron source. Herein, we discuss the challenges and feasibility of deploying a passive neutron source to planetary surfaces for the in situ application of the 40Ar/39Ar chronometer. Requirements in generating and shielding neutrons, as well as analysing samples are described, along with an exploration of limitations such as mass, power and cost. Two potential solutions for the in situ extra-terrestrial deployment of the 40Ar/39Ar method are presented. Although this represents a challenging task, developing the technology to apply the 40Ar/39Ar method on planetary surfaces would represent a major advance towards constraining the timescale of solar system formation and evolution

Quantum systems in weak gravitational fields

Fully covariant wave equations predict the existence of a class of inertial-gravitational effects that can be tested experimentally. In these equations inertia and gravity appear as external classical fields, but, by conforming to general relativity, provide very valuable information on how Einstein's views carry through in the world of the quantum.Comment: 22 pages. To be published in Proceedings of the 17th Course of the International School of Cosmology and Gravitation "Advances in the interplay between quantum and gravity physics" edited by V. De Sabbata and A. Zheltukhin, Kluwer Academic Publishers, Dordrech

A novel link between the proteasome pathway and the signal transduction pathway of the Bone Morphogenetic Proteins (BMPs)

BACKGROUND: The intracellular signaling events of the Bone Morphogenetic Proteins (BMPs) involve the R-Smad family members Smad1, Smad5, Smad8 and the Co-Smad, Smad4. Smads are currently considered to be DNA-binding transcriptional modulators and shown to recruit the master transcriptional co-activator CBP/p300 for transcriptional activation. SNIP1 is a recently discovered novel repressor of CBP/p300. Currently, the detailed molecular mechanisms that allow R-Smads and Co-Smad to co-operatively modulate transcription events are not fully understood. RESULTS: Here we report a novel physical and functional link between Smad1 and the 26S proteasome that contributes to Smad1- and Smad4-mediated transcriptional regulation. Smad1 forms a complex with a proteasome β subunit HsN3 and the ornithine decarboxylase antizyme (Az). The interaction is enhanced upon BMP type I receptor activation and occur prior to the incorporation of HsN3 into the mature 20S proteasome. Furthermore, BMPs trigger the translocation of Smad1, HsN3 and Az into the nucleus, where the novel CBP/p300 repressor protein SNIP1 is further recruited to Smad1/HsN3/Az complex and degraded in a Smad1-, Smad4- and Az-dependent fashion. The degradation of the CBP/p300 repressor SNIP1 is likely an essential step for Smad1-, Smad4-mediated transcriptional activation, since increased SNIP1 expression inhibits BMP-induced gene responses. CONCLUSIONS: Our studies thus add two additional important functional partners of Smad1 into the signaling web of BMPs and also suggest a novel mechanism for Smad1 and Smad4 to co-modulate transcription via regulating proteasomal degradation of CBP/p300 repressor SNIP1