Validation of commercial Mas receptor antibodies for utilization in Western Blotting, immunofluorescence and immunohistochemistry studies

Mas receptor (MasR) is a G protein-coupled receptor proposed as a candidate for mediating the angiotensin (Ang)-converting enzyme 2-Ang (1-7) protective axis of renin-angiotensin system. Because the role of this receptor is not definitively clarified, determination of MasR tissue distribution and expression levels constitutes a critical knowledge to fully understanding its function. Commercially available antibodies have been widely employed for MasR protein localization and quantification, but they have not been adequately validated. In this study, we carried on an exhaustive evaluation of four commercial MasR antibodies, following previously established criteria. Western Blotting (WB) and immunohistochemistry studies starting from hearts and kidneys from wild type (WT) mice revealed that antibodies raised against different MasR domains yielded different patterns of reactivity. Furthermore, staining patterns appeared identical in samples from MasR knockout (MasR-KO) mice. We verified by polymerase chain reaction analysis that the MasR-KO mice used were truly deficient in this receptor as MAS transcripts were undetectable in either heart or kidney from this animal model. In addition, we evaluated the ability of the antibodies to detect the human c-myc-tagged MasR overexpressed in human embryonic kidney cells. Three antibodies were capable of detecting the MasR either by WB or by immunofluorescence, reproducing the patterns obtained with an anti c-myc antibody. In conclusion, although three of the selected antibodies were able to detect MasR protein at high expression levels observed in a transfected cell line, they failed to detect this receptor in mice tissues at physiological expression levels. As a consequence, validated antibodies that can recognize and detect the MasR at physiological levels are still lacking

Non-monotonic variation with salt concentration of the second virial coefficient in protein solutions

The osmotic virial coefficient $B_2$ of globular protein solutions is calculated as a function of added salt concentration at fixed pH by computer simulations of the ``primitive model''. The salt and counter-ions as well as a discrete charge pattern on the protein surface are explicitly incorporated. For parameters roughly corresponding to lysozyme, we find that $B_2$ first decreases with added salt concentration up to a threshold concentration, then increases to a maximum, and then decreases again upon further raising the ionic strength. Our studies demonstrate that the existence of a discrete charge pattern on the protein surface profoundly influences the effective interactions and that non-linear Poisson Boltzmann and Derjaguin-Landau-Verwey-Overbeek (DLVO) theory fail for large ionic strength. The observed non-monotonicity of $B_2$ is compared to experiments. Implications for protein crystallization are discussed.Comment: 43 pages, including 17 figure

New Strong-Field QED Effects at ELI: Nonperturbative Vacuum Pair Production

Since the work of Sauter, and Heisenberg, Euler and K\"ockel, it has been understood that vacuum polarization effects in quantum electrodynamics (QED) predict remarkable new phenomena such as light-light scattering and pair production from vacuum. However, these fundamental effects are difficult to probe experimentally because they are very weak, and they are difficult to analyze theoretically because they are highly nonlinear and/or nonperturbative. The Extreme Light Infrastructure (ELI) project offers the possibility of a new window into this largely unexplored world. I review these ideas, along with some new results, explaining why quantum field theorists are so interested in this rapidly developing field of laser science. I concentrate on the theoretical tools that have been developed to analyze nonperturbative vacuum pair production.Comment: 20 pages, 9 figures; Key Lecture at the ELI Workshop and School on "Fundamental Physics with Ultra-High Fields", 29 Sept - 2 Oct. 2008, Frauenworth Monastery, Germany; v2: refs updated, English translations of reviews of Nikishov and Ritu

Erratum: Antimicrobials: A global alliance for optimizing their rational use in intra-abdominal infections (AGORA). [World J Emerg Surg. 11, (2016) (33)] DOI: 10.1186/s13017-016-0089-y

The original article [1] contains an error whereby a co-author, Boris Sakakushev has their family name spelt incorrectly as 'Sakakhushev'. The authors would therefore like it known that the correct spelling of the family name is 'Sakakushev'. © The Author(s)

Study of fast-ion transport induced by fishbones on JET

The impact of fishbone oscillations onto a confined fast-ion population is simulated for a JET plasma and benchmarked against experiment quantitatively with the help of neutron rate measurements. The transient drops in volume integrated neutron emission are found to be mainly caused by the spatial redistribution of the (neutral beam injected) fast-ion population confined in the plasma rather than by fast-ion loss. The simulations yield a quadratic dependence of the neutron drop on the fishbone amplitude. It is found that the simulations are able to correctly reproduce the magnitude of the experimentally observed drop in volume integrated neutron emission to within a factor 2. Furthermore, frequency chirping is found to be important. Omitting the fishbone frequency chirp in the simulations reduces the magnitude of the neutron rate drop (and hence fast-ion redistribution) to about half its original value