Protocol: High-throughput and quantitative assays of auxin and auxin precursors from minute tissue samples

Background: The plant hormone auxin, indole-3-acetic acid (IAA), plays important roles in plant growth and development. The signaling response to IAA is largely dependent on the local concentration of IAA, and this concentration is regulated by multiple mechanisms in plants. Therefore, the precise quantification of local IAA concentration provides insights into the regulation of IAA and its biological roles. Meanwhile, pathways and genes involved in IAA biosynthesis are not fully understood, so it is necessary to analyze the production of IAA at the metabolite level for unbiased studies of IAA biosynthesis. Results: We have developed high-throughput methods to quantify plant endogenous IAA and its biosynthetic precursors including indole, tryptophan, indole-3-pyruvic acid (IPyA), and indole-3-butyric acid (IBA). The protocol starts with homogenizing plant tissues with stable-labeled internal standards added, followed by analyte purification using solid phase extraction (SPE) tips and analyte derivatization. The derivatized analytes are finally analyzed by selected reaction monitoring on a gas chromatograph-mass spectrometer (GC-MS/MS) to determine the precise abundance of analytes. The amount of plant tissue required for the assay is small (typically 2–10 mg fresh weight), and the use of SPE tips is simple and convenient, which allows preparation of large sets of samples within reasonable time periods. Conclusions: The SPE tips and GC-MS/MS based method enables high-throughput and accurate quantification of IAA and its biosynthetic precursors from minute plant tissue samples. The protocol can be used for measurement of these endogenous compounds using isotope dilution, and it can also be applied to analyze IAA biosynthesis and biosynthetic pathways using stable isotope labeling. The method will potentially advance knowledge of the role and regulation of IAA

Flexible Invariants Through Semantic Collaboration

Modular reasoning about class invariants is challenging in the presence of dependencies among collaborating objects that need to maintain global consistency. This paper presents semantic collaboration: a novel methodology to specify and reason about class invariants of sequential object-oriented programs, which models dependencies between collaborating objects by semantic means. Combined with a simple ownership mechanism and useful default schemes, semantic collaboration achieves the flexibility necessary to reason about complicated inter-object dependencies but requires limited annotation burden when applied to standard specification patterns. The methodology is implemented in AutoProof, our program verifier for the Eiffel programming language (but it is applicable to any language supporting some form of representation invariants). An evaluation on several challenge problems proposed in the literature demonstrates that it can handle a variety of idiomatic collaboration patterns, and is more widely applicable than the existing invariant methodologies.Comment: 22 page

Special Project: Public Housing

Despite the general prosperity of this country, a cursory survey of any American town or city will reveal that many Americans live in housing which is substandard. Frequently one sees unpainted houses characterized by broken windows and inadequate sanitary facilities. In urban areas, the ever present tenement is often filled with too many people and not enough toilets; stairs are dangerous and refuse lies uncollected in the halls. Rooms without windows are common, while those blessed with windows frequently receive little light--the only view is another window of another building. Disease and discomfort are everywhere.These conditions, however, are neither new nor unnoticed.Throughout American history, individuals as well as various levels of government have attempted to remedy, or at least alleviate, the housing problem. This study will explore one such effort--the federally-financed, locally-administered public housing program created by the Housing Act of 1937. Consequently, throughout this study, the term public housing, describing any period after 1937,refers only to the product of this Act, unless otherwise indicated. This study does not concern federally-funded housing programs such as Federal Housing Authority mortgage guarantees, rehabilitation of existing dwellings, and self-help programs

Oral rivaroxaban versus standard therapy for the treatment of symptomatic venous thromboembolism : a pooled analysis of the EINSTEIN-DVT and PE randomized studies

Background: Standard treatment for venous thromboembolism (VTE) consists of a heparin combined with vitamin K antagonists. Direct oral anticoagulants have been investigated for acute and extended treatment of symptomatic VTE; their use could avoid parenteral treatment and/or laboratory monitoring of anticoagulant effects. Methods: A prespecified pooled analysis of the EINSTEIN-DVT and EINSTEIN-PE studies compared the efficacy and safety of rivaroxaban (15 mg twice-daily for 21 days, followed by 20 mg once-daily) with standard-therapy (enoxaparin 1.0 mg/kg twice-daily and warfarin or acenocoumarol). Patients were treated for 3, 6, or 12 months and followed for suspected recurrent VTE and bleeding. The prespecified noninferiority margin was 1.75. Results: 8282 patients were enrolled. 4151 received rivaroxaban and 4131 received standard-therapy. The primary efficacy outcome occurred in 86 rivaroxaban-treated patients (2.1%) compared with 95 (2.3%) standard-therapy-treated patients (hazard ratio, 0.89; 95% confidence interval [CI], 0.66-1.19; pnoninferiority<0.001). Major bleeding was observed in 40 (1.0%) and 72 (1.7%) patients in the rivaroxaban and standard-therapy groups, respectively (hazard ratio, 0.54; 95% CI, 0.37-0.79; p=0.002). In key subgroups, including fragile patients, cancer patients, patients presenting with large clots and those with a history of recurrent VTE, the efficacy and safety of rivaroxaban was similar compared with standard-therapy. Conclusion: The single-drug approach with rivaroxaban resulted in similar efficacy to standard-therapy and was associated with a significantly lower rate of major bleeding. Efficacy and safety results were consistent among key patient subgroups

How efficient are coronal mass ejections at accelerating solar energetic particles?

The largest solar energetic particle (SEP) events are thought to be due to particle acceleration at a shock driven by a fast coronal mass ejection (CME). We investigate the efficiency of this process by comparing the total energy content of energetic particles with the kinetic energy of the associated CMEs. The energy content of 23 large SEP events from 1998 through 2003 is estimated based on data from ACE, GOES, and SAMPEX, and interpreted using the results of particle transport simulations and inferred longitude distributions. CME data for these events are obtained from SOHO. When compared to the estimated kinetic energy of the associated coronal mass ejections (CMEs), it is found that large SEP events can extract ~10% or more of the CME kinetic energy. The largest SEP events appear to require massive, very energetic CMEs

A Unified Account of the Moral Standing to Blame

Recently, philosophers have turned their attention to the question, not when a given agent is blameworthy for what she does, but when a further agent has the moral standing to blame her for what she does. Philosophers have proposed at least four conditions on having “moral standing”: 1. One’s blame would not be “hypocritical”. 2. One is not oneself “involved in” the target agent’s wrongdoing. 3. One must be warranted in believing that the target is indeed blameworthy for the wrongdoing. 4. The target’s wrongdoing must some of “one’s business”. These conditions are often proposed as both conditions on one and the same thing, and as marking fundamentally different ways of “losing standing.” Here I call these claims into question. First, I claim that conditions (3) and (4) are simply conditions on different things than are conditions (1) and (2). Second, I argue that condition (2) reduces to condition (1): when “involvement” removes someone’s standing to blame, it does so only by indicating something further about that agent, viz., that he or she lacks commitment to the values that condemn the wrongdoer’s action. The result: after we clarify the nature of the non-hypocrisy condition, we will have a unified account of moral standing to blame. Issues also discussed: whether standing can ever be regained, the relationship between standing and our "moral fragility", the difference between mere inconsistency and hypocrisy, and whether a condition of standing might be derived from deeper facts about the "equality of persons"

Deriving metallicities from calcium triplet spectroscopy in combination with near-infrared photometry

Context. When they are established with sufficient precision, the ages, metallicities and kinematics of Galactic globular clusters (GGCs) can shed much light on the dynamical and chemical evolution of the Galactic halo and bulge. While the most fundamental way of determining GC abundances is by means of high-resolution spectroscopy, in practice this method is limited to only the brighter stars in the nearest and less reddened objects. This restriction has, over the years, led to the development of a large number of techniques that measure the overall abundance indirectly from parameters that correlate with overall metallicity. One of the most efficient methods is measuring the equivalent width (EW) of the calcium II triplet (CaT) at λ ≈ 8500 Å in red giants, which are corrected for the luminosity and temperature effects using the V magnitude differences from the horizontal branch (HB). Aims. We establish a similar method in the near-infrared (NIR), by combining the power of the differential magnitudes technique with the advantages of NIR photometry to minimize differential reddening effects. Methods. We used the Ks magnitude difference between the star and the reddest part of the HB (RHB) or of the red clump (RC) to generate reduced equivalent widths (rEW) from previously presented datasets. Then we calibrated these rEW against three previously reported different metallicity scales; one of which we corrected using high-resolution spectroscopic metallicities. Results. We calculated the calibration relations for the two datasets and the three metallicity scales and found that they are approximately equivalent, with almost negligible differences. We compared our NIR calibrations with the corresponding optical ones, and found them to be equivalent, which shows that the luminosity-corrected rEW using the Ks magnitude is compatible with the one obtained from the V magnitude. We then used the metallicities obtained from the calibration to investigate the internal metallicity distributions of the GCs. Conclusions. We have established that the ([Fe/H]:rEW) relation is independent of the magnitude used for the luminosity correction and find that the calibration relations change only slightly for different metallicity scales. The CaT technique using NIR photometry is thus a powerful tool to derive metallicities. In particular, it can be used to study the internal metallicity spread of a GC. We confirm the presence of at least two metallicity populations in NGC 6656 and find that several other GCs present peculiar metallicity distributions

Imprints of Short Distance Physics On Inflationary Cosmology

We analyze the impact of certain modifications to short distance physics on the inflationary perturbation spectrum. For the specific case of power-law inflation, we find distinctive -- and possibly observable -- effects on the spectrum of density perturbations.Comment: Revtex 4, 3 eps figs, 4 page

FGFR3 Activates RSK2 to Mediate Hematopoietic Transformation through Tyrosine Phosphorylation of RSK2 and Activation of the MEK/ERK Pathway

SummaryTo better understand the signaling properties of oncogenic FGFR3, we performed phospho-proteomics studies to identify potential downstream signaling effectors that are tyrosine phosphorylated in hematopoietic cells expressing constitutively activated leukemogenic FGFR3 mutants. We found that FGFR3 directly tyrosine phosphorylates the serine/threonine kinase p90RSK2 at Y529, which consequently regulates RSK2 activation by facilitating inactive ERK binding to RSK2 that is required for ERK-dependent phosphorylation and activation of RSK2. Moreover, inhibition of RSK2 by siRNA or a specific RSK inhibitor fmk effectively induced apoptosis in FGFR3-expressing human t(4;14)-positive myeloma cells. Our findings suggest that FGFR3 mediates hematopoietic transformation by activating RSK2 in a two-step fashion, promoting both the ERK-RSK2 interaction and subsequent phosphorylation of RSK2 by ERK