Internalisation of membrane progesterone receptor-α after treatment with progesterone: Potential involvement of a clathrin-dependent pathway

This article has been made available through the Brunel Open Access Publishing Fund.Internalisation and recycling of seven trans-membrane domain receptors is a critical regulatory event for their signalling. The mechanism(s) by which membrane progesterone receptor-α (mPRα) number is regulated on the cell surface is unclear. In this study, we investigated the cellular distribution of mPRα and mechanisms of mPRα trafficking using a cell line derived from a primary culture of human myometrial cells (M11) as an experimental model. RT-PCR and immunofluorescent analysis demonstrated expression of mPRα in M11 cells with mPRα primarily distributed on the cell surface under basal conditions. For the first time, plasma membrane localisation of mPRα was confirmed using immuno-gold transmission electron microscopy. Stimulation of M11 cells with progesterone (P4, 100 nM) resulted in internalisation of mPRα from the plasma membrane to the cytoplasm (10 min) and subsequent partial translocation back to the cell surface (20 min). We investigated potential endocytotic pathways involved in trafficking of mPRα after its internalisation. Partial co-localisation of clathrin with mPRα was obvious after 10 min of P4 treatment. Of note, chlorpromazine (inhibitor of clathrin-mediated pathway) inhibited the endocytosis of mPRα, whereas treatment with nystatin (inhibitor of caveolae-mediated pathway) did not affect internalisation. Collectively, these data suggest that mPRα is expressed on the cell surface of M11 cells and undergoes endocytosis after P4 stimulation primarily via a clathrin-mediated pathway.This article is available through the Brunel Open Access Publishing Fun

HERMIONE: a randomized Phase 2 trial of MM-302 plus trastuzumab versus chemotherapy of physician's choice plus trastuzumab in patients with previously treated, anthracycline-naïve, HER2-positive, locally advanced/metastatic breast cancer.

BackgroundHuman epidermal growth factor receptor 2 (HER2)-positive breast cancer is a particularly aggressive form of the disease, and ultimately progresses in patients with metastases on standard therapies. Anthracyclines, such as doxorubicin, are an effective treatment for HER2-positive breast cancer, particularly when administered in combination with trastuzumab - however, doxorubicin-related cardiotoxicity has limited its use. Many patients are therefore never treated with anthracyclines, even upon disease progression, despite the potential for benefit. MM-302 is a novel, HER2-targeted antibody-liposomal doxorubicin conjugate that specifically targets HER2-overexpressing cells. Preclinical and Phase 1 data suggest that MM-302, as a monotherapy or in combination with trastuzumab, could be effective for managing previously treated, anthracycline-naïve, HER2-positive breast cancer, without the cardiotoxicity observed with free doxorubicin formulations.Methods/designHERMIONE is an open-label, multicenter, randomized (1:1) Phase 2 trial of MM-302 plus trastuzumab versus chemotherapy of physician's choice (gemcitabine, capecitabine, or vinorelbine) plus trastuzumab planned to enroll 250 anthracycline-naïve patients with locally advanced/metastatic HER2-positive breast cancer. Key inclusion criteria are: previous treatment with trastuzumab (with or without pertuzumab) in any setting; refractory or intolerant to pertuzumab (refractory to pertuzumab defined as progression in the locally advanced or metastatic setting, or disease recurrence during or within 12 months of completing pertuzumab-containing neoadjuvant and/or adjuvant therapy); and disease progression on, or intolerant to, ado-trastuzumab emtansine for locally advanced or metastatic disease. The trial is currently being conducted at sites in the USA, Canada, and Western Europe. Treatment will be administered in 21-day cycles, and will be continued until disease progression or unacceptable toxicity. The primary endpoint is independently assessed progression-free survival (PFS). Tumor response will be assessed every 6 weeks, and defined according to RECIST v1.1. Secondary endpoints include investigator-assessed PFS, overall survival (OS), OS rates at 6 months and 1 year, objective response rates, safety and tolerability, quality of life, and the pharmacokinetic profile of MM-302 plus trastuzumab.DiscussionThe HERMIONE study will evaluate the efficacy and safety of MM-302 plus trastuzumab in patients with refractory HER2-positive advanced/metastatic breast cancer for whom there are no standard of care therapies with a proven survival advantage.Trial registrationClinicaltrials.gov identifier: NCT02213744 . Registration date: 06AUG2014

Adaptive Lévy processes and area-restricted search in human foraging

A considerable amount of research has claimed that animals’ foraging behaviors display movement lengths with power-law distributed tails, characteristic of Lévy flights and Lévy walks. Though these claims have recently come into question, the proposal that many animals forage using Lévy processes nonetheless remains. A Lévy process does not consider when or where resources are encountered, and samples movement lengths independently of past experience. However, Lévy processes too have come into question based on the observation that in patchy resource environments resource-sensitive foraging strategies, like area-restricted search, perform better than Lévy flights yet can still generate heavy-tailed distributions of movement lengths. To investigate these questions further, we tracked humans as they searched for hidden resources in an open-field virtual environment, with either patchy or dispersed resource distributions. Supporting previous research, for both conditions logarithmic binning methods were consistent with Lévy flights and rank-frequency methods–comparing alternative distributions using maximum likelihood methods–showed the strongest support for bounded power-law distributions (truncated Lévy flights). However, goodness-of-fit tests found that even bounded power-law distributions only accurately characterized movement behavior for 4 (out of 32) participants. Moreover, paths in the patchy environment (but not the dispersed environment) showed a transition to intensive search following resource encounters, characteristic of area-restricted search. Transferring paths between environments revealed that paths generated in the patchy environment were adapted to that environment. Our results suggest that though power-law distributions do not accurately reflect human search, Lévy processes may still describe movement in dispersed environments, but not in patchy environments–where search was area-restricted. Furthermore, our results indicate that search strategies cannot be inferred without knowing how organisms respond to resources–as both patched and dispersed conditions led to similar Lévy-like movement distributions

Solvent extraction of PDMS tubing as a new method for the capture of volatile organic compounds from headspace

Polydimethylsiloxane (PDMS) tubing is increasingly being used to collect volatile organic compounds (VOCs) from static biological headspace. However, analysis of VOCs collected using PDMS tubing often deploys thermal desorption, where samples are considered as ‘one-offs’ and cannot be used in multiple experiments. In this study, we developed a static headspace VOC collection method using PDMS tubing which is solvent-based, meaning that VOC extracts can be used multiple times and can be linked to biological activity. Using a synthetic blend containing a range of known semiochemicals (allyl isothiocyanate, (Z)-3-hexen-1-ol, 1-octen-3-one, nonanal, (E)-anethol, (S)-bornyl acetate, (E)-caryophyllene and pentadecane) with differing chemical and physicochemical properties, VOCs were collected in static headspace by exposure to PDMS tubing with differing doses, sampling times and lengths. In a second experiment, VOCs from oranges were collected using PDMS sampling of static headspace versus dynamic headspace collection. VOCs were eluted with diethyl ether and analysed using gas chromatography – flame ionization detector analysis and coupled GC – mass spectrometry analysis. GC-FID analysis of collected samples showed that longer PDMS tubes captured significantly greater quantities of compounds than shorter tubes, and that sampling duration significantly altered the recovery of all tested compounds. Moreover, greater quantities of compounds were recovered from closed compared to open systems. Finally, analysis of orange headspace VOCs showed only a few qualitative differences in VOCs recovered compared to dynamic headspace collections, although quantities sampled using PDMS tubing were lower. In summary, extraction of PDMS tubing with diethyl ether solvent captures VOCs from the headspace of synthetic blends and biological samples, and the resulting extracts can be used for multiple experiments linking VOC content to biological activity

A Decision Support Tool for the Pushback Rate Control of Airport Departures

Airport surface congestion control has the potential to mitigate the increase in taxi times and fuel burn at major airports. One possible class of congestion control strategies predicts the departure throughput, and recommends a rate at which to release aircraft pushbacks from the gate. This paper describes the field-testing of these types of strategies at Boston Logan International Airport, focusing on the communication of the suggested rate to the air traffic controller, and additional support for its implementation. Two Android tablet computers were used for the field-tests; one to input the data and the other to display the recommended rate to the air traffic controllers. Two potential decision-support displays were tested: a rate control display that only presented a color-coded suggested pushback rate and a volume control display that provided additional support to the controllers on the number of aircraft that had called-ready and had been released. A survey of controllers showed that they had found the decision-support tool easy to use, especially the additional functionality that is provided by the aircraft volume control display. The field tests were also found to yield significant operational benefits showing that such a congestion control strategy could be effective in practice.United States. Federal Aviation AdministrationNational Science Foundation (U.S.) (Award 0931843

Attributable healthcare utilization and cost of pneumonia due to drug-resistant streptococcus pneumonia: a cost analysis

Background: The burden of disease due to S. pneumoniae (pneumococcus), particularly pneumonia, remains high despite the widespread use of vaccines. Drug resistant strains complicate clinical treatment and may increase costs. We estimated the annual burden and incremental costs attributable to antibiotic resistance in pneumococcal pneumonia. Methods: We derived estimates of healthcare utilization and cost (in 2012 dollars) attributable to penicillin, erythromycin and fluoroquinolone resistance by taking the estimate of disease burden from a previously described decision tree model of pneumococcal pneumonia in the U.S. We analyzed model outputs assuming only the existence of susceptible strains and calculating the resulting differences in cost and utilization. We modeled the cost of resistance from delayed resolution of illness and the resulting additional health services. Results: Our model estimated that non-susceptibility to penicillin, erythromycin and fluoroquinolones directly caused 32,398 additional outpatient visits and 19,336 hospitalizations for pneumococcal pneumonia. The incremental cost of antibiotic resistance was estimated to account for 4% ($91 million) of direct medical costs and 5$233 million) of total costs including work and productivity loss. Most of the incremental medical cost ($82 million) was related to hospitalizations resulting from erythromycin non-susceptibility. Among patients under age 18 years, erythromycin non-susceptibility was estimated to cause 17$38 million in costs, or 39% of pneumococcal pneumonia costs attributable to resistance. Conclusions: We estimate that antibiotic resistance in pneumococcal pneumonia leads to substantial healthcare utilization and cost, with more than one-third driven by macrolide resistance in children. With 5% of total pneumococcal costs directly attributable to resistance, strategies to reduce antibiotic resistance or improve antibiotic selection could lead to substantial savings

Predator-Prey Quasi-cycles from a Path Integral Formalism

The existence of beyond mean field quasi-cycle oscillations in a simple spatial model of predator prey interactions is derived from a path integral formalism. The results agree substantially with those obtained from analysis of similar models using system size expansions of the master equation. In all of these analyses, the discrete nature of predator prey populations and finite size effects lead to persistent oscillations in time, but spatial patterns fail to form. The path integral formalism goes beyond mean field theory and provides a focus on individual realizations of the stochastic time evolution of population not captured in the standard master equation approach.Comment: 4 page

Non-Thermal X-ray Emission from the Northwestern Rim of the Galactic Supernova Remnant G266.2-1.2 (RX J0852.0-4622)

We present a detailed spatially-resolved spectroscopic analysis of two X-ray observations (with a total integration time of 73280 seconds) made of the luminous northwestern rim complex of the Galactic supernova remnant (SNR) G266.2-1.2 (RX J0852.0-4622) with the Chandra X-ray Observatory. G266.2-1.2 is a member of a class of Galactic SNRs which feature X-ray spectra dominated by non-thermal emission: in the cases of these SNRs, the emission is believed to have a synchrotron origin and studies of the X-ray spectra of these SNRs can lend insights into how SNRs accelerate cosmic-ray particles. The Chandra observations have clearly revealed fine structure in this rim complex and the spectra of these features are dominated by non-thermal emission. We have measured the length scales of the upstream structures at eight positions along the rim and derive lengths of 0.02-0.08 pc (assuming a distance of 750 pc to G266.2-1.2). We have also extracted spectra from seven regions in the rim complex and fit these spectra with such models as a simple power law as well as the synchrotron models SRCUT and SRESC. We have constrained our fits to the latter two models using estimates for the flux densities of these filaments at 1 GHz as determined from radio observations made with the Australia Telescope Compact Array (ATCA). Statistically-acceptable fits to all seven regions are derived using each model: differences in the fit parameters (such as photon index and cutoff frequency) are seen in the different regions, which may indicate variations in shock conditions and the maximum energies of the cosmic-ray electrons accelerated at each region. Finally, we estimate the maximum energy of cosmic-ray electrons accelerated along this rim complex to be approximately 40 TeV. We include a summary of estimated maximum energies for both Galactic SNRs as well as SNRs located in the Large Magellanic Cloud.Comment: 56 Pages, 11 Figures, Accepted for publication in the Astrophysical Journa

Fuel Burn and Emissions Reduction Potential of Low Power/Low Drag Approaches

Changing aircraft operational procedures is one strategy that can be used to reduce fuel burn and mitigate environmental impacts of aviation in relatively short timeframes with existing aircraft types. This study quantifies the fuel burn and emissions reduction potential of delayed deceleration approaches, where the aircraft is kept fast and in clean aerodynamic configuration for as long as possible during the approach phase of flight. This reduces the drag and thrust requirements and these procedures are therefore called Low Power/Low Drag approaches. Operational data is used to characterize approach profiles, together with their fuel burn and emissions properties correlated to airspeed and configuration for a selection of aircraft types. Aircraft that were observed to decelerate and configure flaps later in the approach had 30-40% lower fuel burn and carbon dioxide emissions below 10,000 ft compared to those that did not. Estimates of US system-wide fuel burn and emissions reduction potential from Low Power/Low Drag approaches are provided: if only 1% of the total operations used these approaches, savings across all operators would amount to 2.9 million US gallons ($5.8-11.6 million at$2-4/US gallon) of fuel and 28,000 metric tons of carbon dioxide emissions per year. A discussion is provided on the implementation barriers which need to be addressed if benefits are to be realized.United States. Federal Aviation Administration (Award 06-C-NE-MIT, Amendment 017