Structure of the Proline Utilization A Proline Dehydrogenase Domain Inactivated by \u3ci\u3eN\u3c/i\u3e-propargylglycine Provides Insight into Conformational Changes Induced by Substrate Binding and Flavin Reduction

Proline utilization A (PutA) from Escherichia coli is a flavoprotein that has mutually exclusive roles as a transcriptional repressor of the put regulon and a membrane-associated enzyme that catalyzes the oxidation of proline to glutamate. Previous studies have shown that the binding of proline in the proline dehydrogenase (PRODH) active site and subsequent reduction of the FAD trigger global conformational changes that enhance PutA-membrane affinity. These events cause PutA to switch from its repressor to enzymatic role, but the mechanism by which this signal is propagated from the active site to the distal membrane-binding domain is largely unknown. Here, it is shown that N-propargylglycine irreversibly inactivates PutA by covalently linking the flavin N(5) atom to the ε-amino of Lys329. Furthermore, inactivation locks PutA into a conformation that may mimic the proline reduced, membrane-associated form. The 2.15 Å resolution structure of the inactivated PRODH domain suggests that the initial events involved in broadcasting the reduced flavin state to the distal membrane binding domain include major reorganization of the flavin ribityl chain, severe (35 degree) butterfly bending of the isoalloxazine ring, and disruption of an electrostatic network involving the flavin N(5), Arg431, and Asp370. The structure also provides information about conformational changes associated with substrate binding. This analysis suggests that the active site is incompletely assembled in the absence of the substrate, and the binding of proline draws together conserved residues in helix 8 and the β1-αl loop to complete the active site

Astigmatic correction with implantation of a light adjustable vs monofocal lens: a single site analysis of a randomized controlled trial

AIM To evaluate the light adjustable lens (LAL) vs a standard monofocal lens in achieving target astigmatic refraction and improving postoperative uncorrected distance visual acuity (UDVA). METHODS This randomized controlled clinical trial included 40 patients with pre-existing astigmatism and visually significant cataract. Twenty-eight patients received the LAL and 12 control patients received a monofocal intraocular lens (IOL) after cataract extraction at a single institution. The patients with the LAL underwent adjustment by ultraviolet (UV) light postoperatively plus subsequent lock-in procedures and all patients returned to clinic for follow up of study parameters at 6, 9, and 12mo. Manifest refraction, distance visual acuity, and adverse events were recorded at each visit. RESULTS The mean cylinder before adjustment in eyes with the LAL was -0.89±0.58 D (-2.00 to 0.00 D) and -0.34±0.34 D (-1.25 to 0.00 D) after lock-in (P=1.68x10−8). The mean cylinder in patients with the monofocal lens was -1.00±0.32 D (-1.50 to -0.50 D) at 17-21d postoperatively, which was statistically different from the LAL cylinder post lock-in (P=1.43x10−6). UDVA in the LAL group was 20/20 or better in 79% of patients post lock-in with good stability over 12mo compared with 33% of the control patients with UDVA of 20/20 or better. CONCLUSION These results demonstrate that the LAL is more effective in achieving target refractions and improving postoperative UDVA in patients with pre-existing corneal astigmatism than a standard monofocal lens

Calibrating Convective properties of Solar-like Stars in the Kepler Field of View

Stellar models generally use simple parametrizations to treat convection. The most widely used parametrization is the so-called "Mixing Length Theory" where the convective eddy sizes are described using a single number, \alpha, the mixing-length parameter. This is a free parameter, and the general practice is to calibrate \alpha using the known properties of the Sun and apply that to all stars. Using data from NASA's Kepler mission we show that using the solar-calibrated \alpha is not always appropriate, and that in many cases it would lead to estimates of initial helium abundances that are lower than the primordial helium abundance. Kepler data allow us to calibrate \alpha for many other stars and we show that for the sample of stars we have studied, the mixing-length parameter is generally lower than the solar value. We studied the correlation between \alpha and stellar properties, and we find that \alpha increases with metallicity. We therefore conclude that results obtained by fitting stellar models or by using population-synthesis models constructed with solar values of \alpha are likely to have large systematic errors. Our results also confirm theoretical expectations that the mixing-length parameter should vary with stellar properties.Comment: 16 pages, 4 figures, accepted for publication in ApJ

In situ detection of boron by ChemCam on Mars

We report the first in situ detection of boron on Mars. Boron has been detected in Gale crater at levels Curiosity rover ChemCam instrument in calcium-sulfate-filled fractures, which formed in a late-stage groundwater circulating mainly in phyllosilicate-rich bedrock interpreted as lacustrine in origin. We consider two main groundwater-driven hypotheses to explain the presence of boron in the veins: leaching of borates out of bedrock or the redistribution of borate by dissolution of borate-bearing evaporite deposits. Our results suggest that an evaporation mechanism is most likely, implying that Gale groundwaters were mildly alkaline. On Earth, boron may be a necessary component for the origin of life; on Mars, its presence suggests that subsurface groundwater conditions could have supported prebiotic chemical reactions if organics were also present and provides additional support for the past habitability of Gale crater

The advanced LIGO input optics

The advanced LIGO gravitational wave detectors are nearing their design sensitivity and should begin taking meaningful astrophysical data in the fall of 2015. These resonant optical interferometers will have unprecedented sensitivity to the strains caused by passing gravitational waves. The input optics play a significant part in allowing these devices to reach such sensitivities. Residing between the pre-stabilized laser and the main interferometer, the input optics subsystem is tasked with preparing the laser beam for interferometry at the sub-attometer level while operating at continuous wave input power levels ranging from 100 mW to 150 W. These extreme operating conditions required every major component to be custom designed. These designs draw heavily on the experience and understanding gained during the operation of Initial LIGO and Enhanced LIGO. In this article, we report on how the components of the input optics were designed to meet their stringent requirements and present measurements showing how well they have lived up to their design

The Advanced LIGO Input Optics

The advanced LIGO gravitational wave detectors are nearing their design sensitivity and should begin taking meaningful astrophysical data in the fall of 2015. These resonant optical interferometers will have unprecedented sensitivity to the strains caused by passing gravitational waves. The input optics play a significant part in allowing these devices to reach such sensitivities. Residing between the pre-stabilized laser and the main interferometer, the input optics subsystem is tasked with preparing the laser beam for interferometry at the sub-attometer level while operating at continuous wave input power levels ranging from 100 mW to 150 W. These extreme operating conditions required every major component to be custom designed. These designs draw heavily on the experience and understanding gained during the operation of Initial LIGO and Enhanced LIGO. In this article, we report on how the components of the input optics were designed to meet their stringent requirements and present measurements showing how well they have lived up to their design

Control of intestinal stem cell function and proliferation by mitochondrial pyruvate metabolism.

Most differentiated cells convert glucose to pyruvate in the cytosol through glycolysis, followed by pyruvate oxidation in the mitochondria. These processes are linked by the mitochondrial pyruvate carrier (MPC), which is required for efficient mitochondrial pyruvate uptake. In contrast, proliferative cells, including many cancer and stem cells, perform glycolysis robustly but limit fractional mitochondrial pyruvate oxidation. We sought to understand the role this transition from glycolysis to pyruvate oxidation plays in stem cell maintenance and differentiation. Loss of the MPC in Lgr5-EGFP-positive stem cells, or treatment of intestinal organoids with an MPC inhibitor, increases proliferation and expands the stem cell compartment. Similarly, genetic deletion of the MPC in Drosophila intestinal stem cells also increases proliferation, whereas MPC overexpression suppresses stem cell proliferation. These data demonstrate that limiting mitochondrial pyruvate metabolism is necessary and sufficient to maintain the proliferation of intestinal stem cells

Motor phenotype of LRRK2 G2019S carriers in early-onset Parkinson disease

Objective: To determine the motor phenotype of LRRK2 G2019S mutation carriers. LRRK2 mutation carriers were previously reported to manifest the tremor dominant motor phenotype, which has been associated with slower motor progression and less cognitive impairment compared with the postural instability and gait difficulty (PIGD) phenotype. Design: Cross-sectional observational study. Setting: Thirteen movement disorders centers. Participants: Nine hundred twenty-five early-onset Parkinson disease cases defined as age at onset younger than 51 years. Main Outcome Measures: LRRK2 mutation status and Parkinson disease motor phenotype: tremor dominant or PIGD. Demographic information, family history of Parkinson disease, and the Unified Parkinson's Disease Rating Scale score were collected on all participants. DNA samples were genotyped for LRRK2 mutations (G2019S, I2020T, R1441C, and Y1699C). Logistic regression was used to examine associations of G2019S mutation status with motor phenotype adjusting for disease duration, Ashkenazi Jewish ancestry, levodopa dose, and family history of Parkinson disease. Results: Thirty-four cases (3.7%) (14 previously reported) were G2019S carriers. No other mutations were found. Carriers were more likely to be Ashkenazi Jewish (55.9% vs 11.9%; P < .001) but did not significantly differ in any other demographic or disease characteristics. Carriers had a lower tremor score (P = .03) and were more likely to have a PIGD phenotype (92.3% vs 58.9%; P = .003). The association of the G2019S mutation with PIGD phenotype remained after controlling for disease duration and Ashkenazi Jewish ancestry (odds ratio, 17.7; P < .001). Conclusion: Early-onset Parkinson disease G2019S LRRK2 carriers are more likely to manifest the PIGD phenotype, which may have implications for disease course

Decadal-scale litter manipulation alters the biochemical and physical character of tropical forest soil carbon

© 2018 Elsevier Ltd Climate change and rising atmospheric carbon dioxide (CO2) concentrations are likely to alter tropical forest net primary productivity (NPP), potentially affecting soil C storage. We examined biochemical and physical changes in soil C fractions in a humid tropical forest where experimental litter manipulation changed total soil C stocks. We hypothesized that: (1.) low-density soil organic C (SOC) fractions are more responsive to altered litter inputs than mineral-associated SOC, because they cycle relatively rapidly. (2.) Any accumulation of mineral-associated SOC with litter addition is relatively stable (i.e. low leaching potential). (3.) Certain biomolecules, such as waxes (alkyl) and proteins (N-alkyl), form more stable mineral-associations than other biomolecules in strongly weathered soils. A decade of litter addition and removal affected bulk soil C content in the upper 5 cm by +32% and −31%, respectively. Most notably, C concentration in the mineral-associated SOC fraction was greater in litter addition plots relative to controls by 18% and 28% in the dry and wet seasons, respectively, accounting for the majority of greater bulk soil C stock. Radiocarbon and leaching analyses demonstrated that the greater mineral-associated SOC in litter addition plots consisted of new and relatively stable C, with only 3% of mineral-associated SOC leachable in salt solution. Solid-state13C NMR spectroscopy indicated that waxes (alkyl C) and microbial biomass compounds (O-alkyl and N-alkyl C) in mineral-associated SOC are relatively stable, whereas plant-derived compounds (aromatic and phenolic C) are lost from mineral associations on decadal timescales. We conclude that changes in tropical forest NPP will alter the quantity, biochemistry, and stability of C stored in strongly weathered tropical soils