Propagation of CJD Prions in Primary Murine Glia Cells Expressing Human PrPc

There are various existing cell models for the propagation of animal prions. However, in vitro propagation of human prions has been a long-standing challenge. This study presents the establishment of a long-term primary murine glia culture expressing the human prion protein homozygous for methionine at codon 129, which allows in vitro propagation of Creutzfeldt–Jakob disease (CJD) prions (variant CJD (vCJD) and sporadic CJD (sCJD) type MM2). Prion propagation could be detected by Western blotting of pathological proteinase K-resistant prion protein (PrPSc) from 120 days post exposure. The accumulation of PrPSc could be intensified by adding a cationic lipid mixture to the infectious brain homogenate at the time of infection. Stable propagation of human prions in a long-term murine glia cell culture represents a new tool for future drug development and for mechanistic studies in the field of human prion biology. In addition, our cell model can reduce the need for bioassays with human prions and thereby contributes to further implementation of the 3R principles aiming at replacement, reduction and refinement of animal experiments.Peer Reviewe

Evidence-based cross validation for acoustic power transmission for a novel treatment system

Introduction: The novel Trans-Fusimo Treatment System (TTS) is designed to control Magnetic Resonance guided Focused Ultrasound (MRgFUS) therapy to ablate liver tumours under respiratory motion. It is crucial to deliver the acoustic power within tolerance limits for effective liver tumour treatment via MRgFUS. Before application in a clinical setting, evidence of reproducibility and reliability is a must for safe practice. Materials and methods: The TTS software delivers the acoustic power via ExAblate-2100 Conformal Bone System (CBS) transducer. A built-in quality assurance application was developed to measure the force values, using a novel protocol to measure the efficiency for the electrical power values of 100 and 150W for 6s of sonication. This procedure was repeated 30 times by two independent users against the clinically approved ExAblate-2100 CBS for cross-validation. Results: Both systems proved to deliver the power within the accepted efficiency levels (70–90%). Two sample t-tests were used to assess the differences in force values between the ExAblate-2100 CBS and the TTS (p > 0.05). Bland-Altman plots were used to demonstrate the limits of agreement between the two systems falling within the 10% limits of agreement. Two sample t-tests indicated that TTS does not have user dependency (p > 0.05). Conclusions: The TTS software proved to deliver the acoustic power without exceeding the safety levels. Results provide evidence as a part of ISO13485 regulations for CE marking purposes. The developed methodology could be utilised as a part of quality assurance system in clinical settings; when the TTS is used in clinical practice

Analysis to Action: A Guidebook For Conflict-Sensitive USAID Programming in Africa

https://nsuworks.nova.edu/hcas_dcrs_facbooks/1039/thumbnail.jp

Methodology on Quantification of Sonication Duration for Safe Application of MR Guided Focused Ultrasound for Liver Tumour Ablation

Background and objective Magnetic Resonance Guided Focused Ultrasound (MRgFUS) for liver tumour ablation is a challenging task due to motion caused by breathing and occlusion due the ribcage between the transducer and the tumour. To overcome these challenges, a novel system for liver tumour ablation during free breathing has been designed. Methods The novel TRANS-FUSIMO Treatment System (TTS, EUFP7) interacts with a Magnetic Resonance (MR) scanner and a focused ultrasound transducer to sonicate to a moving target in liver. To meet the requirements of ISO 13485; a quality management system for medical device design, the system needs to be tested for certain process parameters. The duration of sonication and, the delay after the sonication button is activated, are among the parameters that need to be quantified for efficient and safe ablation of tumour tissue. A novel methodology is developed to quantify these process parameters. A computerised scope is programmed in LabVIEW to collect data via hydrophone; where the coordinates of fiber-optic sensor assembly was fed into the TRANS-FUSIMO treatment software via Magnetic Resonance Imaging (MRI) to sonicate to the tip of the sensor, which is synchronised with the clock of the scope, embedded in a degassed water tank via sensor assembly holder. The sonications were executed for 50 W, 100 W, 150 W for 10 s to quantify the actual sonication duration and the delay after the emergency stop by two independent operators for thirty times. The deviation of the system from the predefined specs was calculated. Student's-T test was used to investigate the user dependency. Results The duration of sonication and the delay after the sonication were quantified successfully with the developed method. TTS can sonicate with a maximum deviation of 0.16 s (Std 0.32) from the planned duration and with a delay of 14 ms (Std 0.14) for the emergency stop. Student's T tests indicate that the results do not depend on operators (p > .05). Conclusion The evidence obtained via this protocol is crucial for translation- of-research into the clinics for safe application of MRgFUS. The developed protocol could be used for system maintenance in compliance with quality systems in clinics for daily quality assurance routines

Studying the Physical Diversity of Late-M Dwarfs with Dynamical Masses

We present a systematic study of the physical properties of late-M dwarfs based on high-quality dynamical mass measurements and near-infrared (NIR) spectra. We use astrometry from Keck NGS and LGS AO imaging to determine orbits for late-M binaries. We find that LP 349-25 (M7.5+M8) is a pair of young brown dwarfs (Mtot = 0.120 Msun) for which Lyon and Tucson evolutionary models jointly predict an age of 140+/-30 Myr. This is consistent with the age of the Pleiades, but at least LP 349-25A defies the empirical Pleiades lithium depletion boundary, implying that the system is in fact older and that evolutionary models underpredict the component luminosities. We find that LHS 1901AB (M6.5+M6.5) is a pair of very low-mass stars (Mtot = 0.194 Msun) with model-derived ages consistent with limits from its lack of activity (> 6 Gyr). Our improved orbit for Gl 569Bab (M8.5+M9) results in a higher mass for this binary (Mtot = 0.140 Msun) compared to previous work (0.125 Msun). We use these masses along with our published results for 2MASS J2206-2047AB (M8+M8) to test four sets of ultracool model atmospheres currently in use. Fitting these models to our NIR integrated-light spectra provides temperature estimates warmer by ~250 K than those derived independently from Dusty evolutionary models given the measured masses and luminosities. We propose that model atmospheres are more likely to be the source of this discrepancy, as it would be difficult to explain a uniform temperature offset over such a wide range of masses, ages, and activity levels in the context of evolutionary models. Our results contrast those of Konopacky et al. as we find an opposite and smaller mass discrepancy from what they report when we adopt their model-testing approach since our Teff estimates from fitting spectra are ~650 K higher than from their fitting of broadband photometry alone.Comment: 53 pages, 12 figures, accepted to Ap

Determination of the number of light neutrino species from single photon production at LEP

A determination of the number of light neutrino families performed by measuring the cross section of single photon production in \ee\ collision near the \Zo\ resonance is reported. From an integrated luminosity of $100~\mathrm{pb^{-1}}$, collected during the years 1991--94, we have observed 2091 single photon candidates with an energy above 1~\GeV\ in the polar angular region $45^\circ < \theta_\gamma < 135^\circ$. From a maximum likelihood fit to the single photon cross section, the \Zo\ decay width into invisible particles is measured to be \Ginv = 498 \pm 12 \mathrm{(stat)} \pm 12 \mathrm{(sys)~MeV}. Using the Standard Model couplings of neutrinos to the \Zo, the number of light neutrino species is determined to be $N_\nu = 2.98 \pm 0.07 (\mathrm{stat}) \pm 0.07 (\mathrm{sys}).

Search for R-Parity Breaking Sneutrino Exchange at LEP

We report on a search for R--parity breaking effects due to supersymmetric tau--sneutrino exchange in the reactions e+e- to e+e- and e+e- to mu+mu- at centre--of--mass energies from 91~{\GeV} to 172~{\GeV}, using the L3 detector at LEP. No evidence for deviations from the Standard Model expectations of the measured cross sections and forward--backward asymmetries for these reactions is found. Upper limits for the couplings $\lambda_{131}$ and $\lambda_{232}$ for sneutrino masses up to m_{\SNT} \leq 190~\GeV are determined from an analysis of the expected effects due to tau sneutrino exchange