Nerve growth factor induces neurite outgrowth of PC12 cells by promoting Gβγ-microtubule interaction

Background: Assembly and disassembly of microtubules (MTs) is critical for neurite outgrowth and differentiation. Evidence suggests that nerve growth factor (NGF) induces neurite outgrowth from PC12 cells by activating the receptor tyrosine kinase, TrkA. G protein-coupled receptors (GPCRs) as well as heterotrimeric G proteins are also involved in regulating neurite outgrowth. However, the possible connection between these pathways and how they might ultimately converge to regulate the assembly and organization of MTs during neurite outgrowth is not well understood. Results: Here, we report that Gβγ, an important component of the GPCR pathway, is critical for NGF-induced neuronal differentiation of PC12 cells. We have found that NGF promoted the interaction of Gβγ with MTs and stimulated MT assembly. While Gβγ-sequestering peptide GRK2i inhibited neurite formation, disrupted MTs, and induced neurite damage, the Gβγ activator mSIRK stimulated neurite outgrowth, which indicates the involvement of Gβγ in this process. Because we have shown earlier that prenylation and subsequent methylation/demethylation of γ subunits are required for the Gβγ-MTs interaction in vitro, small-molecule inhibitors (L-28 and L-23) targeting prenylated methylated protein methyl esterase (PMPMEase) were tested in the current study. We found that these inhibitors disrupted Gβγ and ΜΤ organization and affected cellular morphology and neurite outgrowth. In further support of a role of Gβγ-MT interaction in neuronal differentiation, it was observed that overexpression of Gβγ in PC12 cells induced neurite outgrowth in the absence of added NGF. Moreover, overexpressed Gβγ exhibited a pattern of association with MTs similar to that observed in NGF-differentiated cells. Conclusions: Altogether, our results demonstrate that βγ subunit of heterotrimeric G proteins play a critical role in neurite outgrowth and differentiation by interacting with MTs and modulating MT rearrangement. Electronic supplementary material The online version of this article (doi:10.1186/s12868-014-0132-4) contains supplementary material, which is available to authorized users

Accurate gamma and MeV-electron track reconstruction with an ultra-low diffusion Xenon/TMA TPC at 10 atm

We report the performance of a 10 atm Xenon/trimethylamine time projection chamber (TPC) for the detection of X-rays (30 keV) and gamma-rays (0.511-1.275 MeV) in conjunction with the accurate tracking of the associated electrons. When operated at such a high pressure and in similar to 1%-admixtures, trimethylamine (TMA) endows Xenon with an extremely low electron diffusion (1.3 +/- 0.13 mm-sigma (longitudinal), 0.95 +/- 0.20 mm-sigma (transverse) along 1 m drift) besides forming a convenient Penning-Fluorescent' mixture. The TPC, that houses 1.1 kg of gas in its fiducial volume, operated continuously for 100 live-days in charge amplification mode. The readout was performed through the recently introduced microbulk Micromegas technology and the AFTER chip, providing a 3D voxelization of 8 mm x 8 mm x 1.2 mm for approximately 10 cm/MeV-long electron tracks. Resolution in energy (epsilon) at full width half maximum (R) inside the fiducial volume ranged from R = 14.6% (30 keV) to R = 4.6% (1.275 MeV). This work was developed as part of the R&D program of the NEXT collaboration for future detector upgrades in the search of the neutrino-less double beta decay (beta beta 0 nu) in Xe-136, specifically those based on novel gas mixtures. Therefore we ultimately focus on the calorimetric and topological properties of the reconstructed MeV-electron tracks. In particular, the obtained energy resolution has been decomposed in its various contributions and improvements towards achieving the R =1.4%root MeV/epsilon levels obtained in small sensors are discussedThe NEXT collaboration acknowledges funding support from the following agencies and institutions: European Research Council under Advanced Grant 339787-NEXT and Starting Grant 240054-TREX, Spanish Ministerio de Economia y Competitividad under grants Consolider-Ingenio 2010 CSD2008-0037 (CUP) and CSD2007-00042 (CPAN), contracts FPA2008-03456 and FPA2009-13697; Portuguese Fundacao para a Ciencia e a Tecnologia; European FEDER under grant PPTDC/FIS/103860/2008; US Department Of Energy under contract DE-AC02-05CH11231.Gonzalez Diaz, D.; Álvarez Puerta, V.; Borges, FIG.; Camargo, M.; Carcel, S.; Cebrian, S.; Cervera, A.... (2015). Accurate gamma and MeV-electron track reconstruction with an ultra-low diffusion Xenon/TMA TPC at 10 atm. Nuclear Instruments and Methods in Physics Research Section A: Accelerators, Spectrometers, Detectors and Associated Equipment. 804:8-24. https://doi.org/10.1016/j.nima.2015.08.033S82480

Prospective Observational Study of Pazopanib in Patients with Advanced Renal Cell Carcinoma (PRINCIPAL Study)

Background: Real-world data are essential to accurately assessing efficacy and toxicity of approved agents in everyday practice. PRINCIPAL, a prospective, observational study, was designed to confirm the real-world safety and efficacy of pazopanib in patients with advanced renal cell carcinoma (RCC). Subjects, Materials, and Methods: Patients with clear cell advanced/metastatic RCC and a clinical decision to initiate pazopanib treatment within 30 days of enrollment were eligible. Primary objectives included progression-free survival (PFS), overall survival (OS), objective response rate (ORR), relative dose intensity (RDI) and its effect on treatment outcomes, change in health-related quality of life (HRQoL), and safety. We also compared characteristics and outcomes of clinical-trial-eligible (CTE) patients, defined using COMPARZ trial eligibility criteria, with those of non-clinical-trial-eligible (NCTE) patients. Secondary study objectives were to evaluate clinical efficacy, safety, and RDI in patient subgroups. Results: Six hundred fifty-seven patients were enrolled and received ≥1 dose of pazopanib. Median PFS and OS were 10.3 months (95% confidence interval [CI], 9.2–12.0) and 29.9 months (95% CI, 24.7 to not reached), respectively, and the ORR was 30.3%. HRQoL showed no or little deterioration over time. Treatment-related serious adverse events (AEs) and AEs of special interest occurred in 64 (9.7%), and 399 (60.7%) patients, respectively. More patients were classified NCTE than CTE (85.2% vs. 14.8%). Efficacy of pazopanib was similar between the two groups. Conclusion: PRINCIPAL confirms the efficacy and safety of pazopanib in patients with advanced/metastatic RCC in a real-world clinical setting. Implications for Practice: PRINCIPAL is the largest (n = 657) prospective, observational study of pazopanib in patients with advanced/metastatic renal cell carcinoma, to the authors’ knowledge. Consistent with clinical trial results that often contain specific patient types, the PRINCIPAL study demonstrated that the effectiveness and safety of pazopanib is similarly safe and effective in patients with advanced kidney cancer in a real-world clinical setting. The PRINCIPAL study showed that patients with advanced kidney cancer who are treated with first-line pazopanib generally do not show disease progression for approximately 10 months and generally survive for nearly 30 months

Radio Frequency and DC High Voltage Breakdown of High Pressure Helium, Argon, and Xenon

[EN] Motivated by the possibility of guiding daughter ions from double beta decay events to single-ion sensors for barium tagging, the NEXT collaboration is developing a program of R&D to test radio frequency (RF) carpets for ion transport in high pressure xenon gas. This would require carpet functionality in regimes at higher pressures than have been previously reported, implying correspondingly larger electrode voltages than in existing systems. This mode of operation appears plausible for contemporary RF-carpet geometries due to the higher predicted breakdown strength of high pressure xenon relative to low pressure helium, the working medium in most existing RF carpet devices. In this paper we present the first measurements of the high voltage dielectric strength of xenon gas at high pressure and at the relevant RF frequencies for ion transport (in the 10MHz range), as well as new DC and RF measurements of the dielectric strengths of high pressure argon and helium gases at small gap sizes. We find breakdown voltages that are compatible with stable RF carpet operation given the gas, pressure, voltage, materials and geometry of interest.Woodruff, K.; Baeza-Rubio, J.; Huerta, D.; Jones, BJP.; Mcdonald, AD.; Norman, L.; Nygren, DR.... (2020). Radio Frequency and DC High Voltage Breakdown of High Pressure Helium, Argon, and Xenon. Journal of Instrumentation. 15(4):1-15. https://doi.org/10.1088/1748-0221/15/04/P04022S115154Dehmelt, H. G., & Major, F. G. (1962). Orientation of(He4)+Ions by Exchange Collisions with Cesium Atoms. 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