Selected Reaction Monitoring (SRM) frente a Western Blot

We propose the SRM technology as a complementary method to the Western Blot for the detection and quantification of proteins in a sample. The technique Western Blot has its own limitations: i) only a protein-of-choice is detected, ignoring any non-relevant proteins, ii) the sensitivity of the technique depends on the specificity of the antibody and iii) Western Blot is expensive and time-consuming. The advantages of SRM with respect Western Blot are remarkable: i) you can detect up to hundreds of different proteins in a sample, ii) SRM is more sensitive, because just 50 copies of the target protein per cell are enough for the detection and iii) once it has been made an investment in the necessary machinery to develop this technique, the detection of proteins in a sample turns into a cheaper, faster, more specific and full-quantitative procedure, without the need of using antibodies. First of all, SRM requires the identification of little peptides, obtained by tryptic digestion, whose sequence must be unique for a single protein or isoform. There is software for that aim. Then, it’s necessary to create isotope-labeled peptides of that identified for acting as internal standards. That sample is introduced in a triple quadrupole mass spectrometer: it passes through a first quadrupole, which functions as a filter, where the fragments are selected, previously ionized, attending to the mass/charge (m/z) relation that correspond to that unique fragments of the protein of interest. In this first selection may be other peptides from other proteins, with the same m/z but with different sequence. To select those that are exclusive from the target protein, the fragments are moved to a second quadrupole, where they are fragmented again with a physical method, and so new smaller fragments are generated. All the new fragments are conduced to the third quadrupole, where just those which come from the protein of interest are selected, attending at their m/z again. The target peptide concentration is determined by measuring the observed signal response for the target peptide relative to that of the isotopic-labeled peptide, the concentration of which is calculated from a pre-determined calibration-response curve. Calibration curves have to be generated for each target peptide in the sample. Because SRM technology is increasing its use, there have been developed databases where the scientific community upload information about protocols and standards for each protein with the aim to facilitate the work to other researchers.

Evaluating the responsiveness of accelerometry to detect change in physical activity

The responsiveness to change of the Actical and ActiGraph accelerometers was assessed in children and adolescents. Participants (n=208) aged 6-16 years completed two simulated free-living protocols, one with primarily light-to-moderate physical activities (PA) and one with mostly moderate-to-vigorous PA. Time in sedentary, light, moderate, and vigorous PA was estimated using 8 previously developed cut-points (4 for Actical and 4 for ActiGraph) and 15-s and 30-s epochs. Accelerometer responsiveness for detecting differences in PA between protocols was assessed using standardized response means (SRM). SRM values >/=0.8 represented high responsiveness to change. Both accelerometers showed high responsiveness for all PA intensities (SRMs = 1.2-4.7 for Actical and 1.1-3.3 for ActiGraph). All cut-points and epoch lengths yielded high responsiveness, and choice of cut-points and epoch length had little effect on responsiveness. Thus, both the Actical and ActiGraph can detect change in PA in a simulated free-living setting, irrespective of cut-point selection or epoch length.

Measuring Competence in Supervisees and Supervisors : Satisfaction and Related Reactions in Supervision

Supervision is a highly valued component of practitioner training. This chapter discusses the following: factors influencing perceived satisfaction and alliance; and how satisfaction, alliance, and supervision relationships are currently measured; and reviews issues with the concept and its assessment. Given the importance of the supervisory relationship and of the supervisory alliance for the effectiveness of supervision and for the welfare of supervisees, the routine, repeated measurement of both these concepts, together with supervisee satisfaction, also assumes considerable utility. The chapter describes a selection of some commonly used measures: Supervisee Satisfaction Questionnaire (SSQ), Supervisory Relationship Questionnaire (SRQ), Supervisory Relationship Measure (SRM), Supervision Attitude Scale (SAS), Supervisory Working Alliance Inventory (SWAI), Supervisory Styles Inventory (SSI), Role Conflict and Ambiguity Inventory (RCAIC), and Evaluation Process within Supervision Inventory (EPSI).

Analysis of Daphnane Orthoesters in Poisonous Australian Pimelea Species by Liquid Chromatography-Tandem Mass Spectrometry.

Cattle grazing in arid rangelands of Australia suffer periodic extensive and serious poisoning by the plant species Pimelea trichostachya, P. simplex, and P. elongata. Pimelea poisoning (also known as St. George disease and Marree disease) has been attributed to the presence of the diterpenoid orthoester simplexin in these species. However, literature relating to previous studies is complicated by taxonomic revisions, and the presence of simplexin has not previously been verified in all currently recognized taxa capable of inducing pimelea poisoning syndrome, with no previous chemical studies of P. trichostachya (as currently classified) or P. simplex subsp. continua. We report here the isolation of simplexin from P. trichostachya and the development of a liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS) method to measure simplexin concentrations in pimelea plant material. Simplexin was quantified by positive-ion atmospheric pressure chemical ionization (APCI) LC-MS/MS with selected reaction monitoring (SRM) of the m/z 533.3 > 253.3 transition. LC-MS/MS analysis of the four poisonous taxa P. trichostachya, P. elongata, P. simplex subsp. continua, and P. simplex subsp. simplex showed similar profiles with simplexin as the major diterpenoid ester component in all four taxa accompanied by varying amounts of related orthoesters. Similar analyses of P. decora, P. haematostachya, and P. microcephala also demonstrated the presence of simplexin in these species but at far lower concentrations, consistent with the limited reports of stock poisoning associated with these species. The less common, shrubby species P. penicillaris contained simplexin at up to 55 mg/kg dry weight and would be expected to cause poisoning if animals consumed sufficient plant material.

Support vector machine for evaluating seismic-liquefaction potential using shear wave velocity

The use of the shear wave velocity data as a field index for evaluating the liquefaction potential of sands is receiving increased attention because both shear wave velocity and liquefaction resistance are similarly influenced by many of the same factors such as void ratio, state of stress, stress history and geologic age. In this paper, the potential of support vector machine (SVM) based classification approach has been used to assess the liquefaction potential from actual shear wave velocity data. In this approach, an approximate implementation of a structural risk minimization (SRM) induction principle is done, which aims at minimizing a bound on the generalization error of a model rather than minimizing only the mean square error over the data set. Here SVM has been used as a classification tool to predict liquefaction potential of a soil based on shear wave velocity. The dataset consists the information of soil characteristics such as effective vertical stress (sigma'(v0)), soil type, shear wave velocity (V-s) and earthquake parameters such as peak horizontal acceleration (a(max)) and earthquake magnitude (M). Out of the available 186 datasets, 130 are considered for training and remaining 56 are used for testing the model. The study indicated that SVM can successfully model the complex relationship between seismic parameters, soil parameters and the liquefaction potential. In the model based on soil characteristics, the input parameters used are sigma'(v0), soil type. V-s, a(max) and M. In the other model based on shear wave velocity alone uses V-s, a(max) and M as input parameters. In this paper, it has been demonstrated that Vs alone can be used to predict the liquefaction potential of a soil using a support vector machine model. (C) 2010 Elsevier B.V. All rights reserved.

High Temperature Phase Transition Studies and Dielectric Properties of Sr2SbMnO6

Sr2SbMnO6 (SSMO) ceramics were, fabricated using the nanocrystalline powders obtained via molten salt synthesis (MSS) method. High temperature X-ray diffraction studies confirmed the structural phase transition (room temperature tetragonal (I4/mcm) to the cubic phase (Pm-3m)) temperature to be around 736K. The discontinuity in the phase transition indicated its first order nature reflecting the presence of ferroelectric-like distortions in SSMO prepared from MSS which seemed to be unique as it was not observed so far in the case of SSMO prepared using solid-state reaction method. The dielectric behavior of SSMO was studied in the 300-950 K temperature range at high frequencies (MHz range) in order to suppress the of space charge and related effects that dominate at such higher temperatures and mask the real phase transition.

Optical Properties Study of S40Se60-xSbx Bulk Alloys

Bulk samples of S40Se60-xSbx (with x=10, 20, 30, 40 at. %) were prepared from high purity S, Se and Sb by melt quenching method. XRD studies revealed that all the samples have poly-crystalline phase. The variation in optical properties with composition has been investigated by XPS and Raman spectroscopy. The intensity of XPS core level spectra changes with addition of Sb clearly interprets the optical properties change due to compositional variation. The Raman shift and new peak formation in these samples clearly shows the structural modifications due to Sb addition.

Photo induced Effects on the Optical Properties of Sb40Se20S40 Thin Film

The thermally evaporated amorphous Sb40Se20S40 thin film of 800 nm thickness was subjected to light exposure for photo induced studies. The as-prepared and illuminated thin films were studied by X-ray diffraction, Fourier Transform Infrared Spectroscopy and X-ray Photoelectron Spectroscopy. The optical band gap was reduced due to photo induced effects along with the increase in disorder. These optical properties changes are due to the change of homopolar bond densities. The core level peak shifting in XPS spectra supports the optical changes happening in the film due to light exposure.

Nonlinear Refraction and Two-Photon Absorption in Dense 2Bi(2)O(3)-B2O3 Glasses

High density transparent glasses (7.86 g/cc) were fabricated in the 2Bi(2)O(3)-B2O3 (BBO) system. Optical band gap of the obtained glasses was found to be 2.6eV. The refractive index measured for these glasses was 2.25 +/- 0.05 at lambda=543 nm. Nonlinear refraction and absorption studies were carried out on the BBO glasses using z-scan technique a lambda=532 nm of 10 ns pulse width. The nonlinear refractive index obtained was n(2)=12.1x10(-14) cm(2)/W and nonlinear absorption coefficient was beta=15.2 cm/GW. The n(2) and beta values of the BBO glasses were large compared to the other reported high index bismuth based oxide glass systems in the literature. These were attributed to the high density, high linear refractive index, low band gap and two photon absorption associated with these glasses. The electronic origin of large nonlinearities was discussed based on bond-orbital theory.

Effect of Thickness on The Optical Properties Change in Bi/As2S3 Bilayer Thin Films

Bilayer thin films of Bi/As2S3 were prepared from Bi and As2S3 by thermal evaporation technique under high vacuum. We have prepared three bilayer films of 905nm, 910nm and 915nm thickness with with As2S3 as bottom layer (900nm) and Bi as top layer (5,10,15 nm). We have compared the optical changes due to the thickness variation of Bi layer on As2S3 film. The changes were characterized by FTIR and XPS techniques.

Effect of Oxygen Pressure on Structural and Magnetic Properties of YIG Thin Films

The effect of oxygen pressure (P-O2) on the Yttrium Iron Garnet (YIG) thin films were grown on silicon substrate by rf sputtering method was studied. The as-deposited films at 300K were amorphous in nature. The crystallization of these films was achieved by annealing at a temperature of 800 degrees C/1hr in air. The structural, microstructural and magnetic properties were found to be dependent on P-O2.

Raman & Infrared Study of Bi Filled Co4Sb12

CoSb3 skutterudites are established thermoelectric materials in the 500-800K temperature range. Undoped and Bi filled CoSb3 samples were synthesized by induction melting-annealing process and phase confirmation done by X-Ray diffraction. The role of bismuth as a filler in CoSb3 was investigated by Raman and far infrared reflectance study. It was found that bismuth strengthens Sb vibrations, and can potentially scatter Sb related acoustic phonons effectively. As a result substantial reduction in thermal conductivity may be possible with proper control of Bi filling.

Science of geoengineering

Carbon dioxide emissions from the burning of coal, oil, and gas are increasing atmospheric carbon dioxide concentrations. These increased concentrations cause additional energy to be retained in Earth's climate system, thus increasing Earth's temperature. Various methods have been proposed to prevent this temperature increase either by reflecting to space sunlight that would otherwise warm Earth or by removing carbon dioxide from the atmosphere. Such intentional alteration of planetary-scale processes has been termed geoengineering. The first category of geoengineering method, solar geoengineering (also known as solar radiation management, or SRM), raises novel global-scale governance and environmental issues. Some SRM approaches are thought to be low in cost, so the scale of SRM deployment will likely depend primarily on considerations of risk. The second category of geoengineering method, carbon dioxide removal (CDR), raises issues related primarily to scale, cost, effectiveness, and local environmental consequences. The scale of CDR deployment will likely depend primarily on cost.

Sensitivity of simulated climate to latitudinal distribution of solar insolation reduction in solar radiation management

Solar radiation management (SRM) geoengineering has been proposed as a potential option to counteract climate change. We perform a set of idealized geoengineering simulations using Community Atmosphere Model version 3.1 developed at the National Center for Atmospheric Research to investigate the global hydrological implications of varying the latitudinal distribution of solar insolation reduction in SRM methods. To reduce the solar insolation we have prescribed sulfate aerosols in the stratosphere. The radiative forcing in the geoengineering simulations is the net forcing from a doubling of CO2 and the prescribed stratospheric aerosols. We find that for a fixed total mass of sulfate aerosols (12.6 Mt of SO4), relative to a uniform distribution which nearly offsets changes in global mean temperature from a doubling of CO2, global mean radiative forcing is larger when aerosol concentration is maximum at the poles leading to a warmer global mean climate and consequently an intensified hydrological cycle. Opposite changes are simulated when aerosol concentration is maximized in the tropics. We obtain a range of 1 K in global mean temperature and 3% in precipitation changes by varying the distribution pattern in our simulations: this range is about 50% of the climate change from a doubling of CO2. Hence, our study demonstrates that a range of global mean climate states, determined by the global mean radiative forcing, are possible for a fixed total amount of aerosols but with differing latitudinal distribution. However, it is important to note that this is an idealized study and thus not all important realistic climate processes are modeled.