Antibody-based detection of protein phosphorylation status to track the efficacy of novel therapies using nanogram protein quantities from stem cells and cell lines

This protocol describes a highly reproducible antibody-based method that provides protein level and phosphorylation status information from nanogram quantities of protein cell lysate. Nanocapillary isoelectric focusing (cIEF) combines with UV-activated linking chemistry to detect changes in phosphorylation status. As an example application, we describe how to detect changes in response to tyrosine kinase inhibitors (TKIs) in the phosphorylation status of the adaptor protein ​CrkL, a major substrate of the oncogenic tyrosine kinase ​BCR-​ABL in chronic myeloid leukemia (CML), using highly enriched CML stem cells and mature cell populations in vitro. This protocol provides a 2.5 pg/nl limit of protein detection (<0.2% of a stem cell sample containing <104 cells). Additional assays are described for phosphorylated tyrosine 207 (pTyr207)-​CrkL and the protein tyrosine phosphatase ​PTPRC/​CD45; these assays were developed using this protocol and applied to CML patient samples. This method is of high throughput, and it can act as a screen for in vitro cancer stem cell response to drugs and novel agents

Are Electrons Oscillating Photons, Oscillating “Vacuum," or Something Else? The 2015 Panel Discussion: An Unprecedented Engineering Opportunity: A Dynamical Linear Theory of Energy as Light and Matter

Platform: What physical attributes separate EM waves, of the enormous band of radio to visible to x-ray, from the high energy narrow band of gamma-ray? From radio to visible to x-ray, telescopes are designed based upon the optical imaging theory; which is an extension of the Huygens-Fresnel diffraction integral. Do we understand the physical properties of gamma rays that defy us to manipulate them similarly? One demonstrated unique property of gamma rays is that they can be converted to elementary particles (electron and positron pair); or a particle-antiparticle pair can be converted into gamma rays. Thus, EM waves and elementary particles, being inter-convertible; we cannot expect to understand the deeper nature of light without succeeding to find structural inter-relationship between photons and particles. This topic is directly relevant to develop a deeper understanding of the nature of light; which will, in turn, help our engineers to invent better optical instruments

16-O-methylcafestol is present in ground roast Arabica coffees: Implications for authenticity testing

High-field and low-field proton NMR spectroscopy were used to analyse lipophilic extracts from ground roast coffees. Using a sample preparation method that produced concentrated extracts, a small marker peak at 3.16 ppm was observed in 30 Arabica coffees of assured origin. This signal has previously been believed absent from Arabicas, and has been used as a marker for detecting adulteration with robusta. Via 2D 600 MHz NMR and LC-MS, 16-O-methylcafestol and 16-O-methylkahweol were detected for the first time in Arabica roast coffee and shown to be responsible for the marker peak. Using low-field NMR, robusta in Arabica could be detected at levels of the order of 1-2% w/w. A surveillance study of retail purchased "100% Arabica" coffees found that 6 out of 60 samples displayed the 3.16 ppm marker signal to a degree commensurate with adulteration at levels of 3-30% w/w

Smell-related quality of life changes after total laryngectomy : a multi-centre study

Purpose: A total laryngectomy creates an alternate airway for gas exchange that bypasses the upper aerodigestive tract. The subsequent reduction in nasal airflow, and therefore, reduction in deposition of particles to the olfactory neuroepithelium leads to hyposmia or anosmia. The aim of this study was to assess the quality of life impairment conferred by anosmia following laryngectomy and identify any specific patient-related risk factors that are associated with poorer outcomes. Methods: Consecutive patients with a total laryngectomy presenting for review at three tertiary head and neck services (in Australia, the United Kingdom and India) over a 12-month period were recruited. Patient demographic and clinical data were collected, and each subject completed the validated assessment of self-reported olfactory functioning and olfaction-related quality of life questionnaire (ASOF). Dichotomous comparisons were performed using the student's unpaired t-test for continuous variables (SRP), a chi-squared test for categorical variables, and a Kendall's tau-b for ordinal variables (SOC) to assess for a correlation with poorer questionnaire scores. Results: A total of 66 laryngectomees (13.4% female; age 65.7 ± 8.6 years) were included in the study. The mean SRP score of the cohort was found to be 15.6 ± 7.4, while the mean ORQ score was noted to be 16.4 ± 8.1. No other specific risk factors associated with poorer quality of life were identified. Conclusion: A significant quality of life detriment from hyposmia is conferred following laryngectomy. Further research to assess treatment options and the patient population that would best benefit from these interventions is required

Distinguishing the nature of comparable-mass neutron star binary systems with multimessenger observations: GW170817 case study

The discovery of GW170817 with gravitational waves (GWs) and electromagnetic (EM) radiation is prompting new questions in strong-gravity astrophysics. Importantly, it remains unknown whether the progenitor of the merger comprised two neutron stars (NSs), or a NS and a black hole (BH). Using new numerical-relativity simulations and incorporating modeling uncertainties we produce novel GW and EM observables for NS-BH mergers with similar masses. A joint analysis of GW and EM measurements reveals that if GW170817 is a NS-BH merger, <40% of the binary parameters consistent with the GW data are compatible with EM observations.Comment: 8 page

Multimessenger search for sources of gravitational waves and high-energy neutrinos: Initial results for LIGO-Virgo and IceCub

We report the results of a multimessenger search for coincident signals from the LIGO and Virgo gravitational-wave observatories and the partially completed IceCube high-energy neutrino detector, including periods of joint operation between 2007–2010. These include parts of the 2005–2007 run and the 2009–2010 run for LIGO-Virgo, and IceCube’s observation periods with 22, 59 and 79 strings. We find no significant coincident events, and use the search results to derive upper limits on the rate of joint sources for a range of source emission parameters. For the optimistic assumption of gravitational-wave emission energy of 10−2  M⊙c2 at ∼150  Hz with ∼60  ms duration, and high-energy neutrino emission of 1051  erg comparable to the isotropic gamma-ray energy of gamma-ray bursts, we limit the source rate below 1.6×10−2  Mpc−3 yr−1. We also examine how combining information from gravitational waves and neutrinos will aid discovery in the advanced gravitational-wave detector era