Targeted knock‐in (KI) can be achieved in embryos by clustered regularly interspaced short palindromic repeats (CRISPR)‐assisted homology directed repair (HDR). However, HDR efficiency is constrained ...by the competition of nonhomologous end joining. The objective of this study was to explore whether CRISPR‐assisted targeted KI rates can be improved in bovine embryos by exposure to the HDR enhancer RS‐1. In vitro produced zygotes were injected with CRISPR components (300 ng/µl Cas9 messenger RNA and 100 ng/µl single guide RNA against a noncoding region) and a single‐stranded DNA (ssDNA) repair template (100 ng/µl). ssDNA template contained a 6 bp XbaI site insert, allowing targeted KI detection by restriction analysis, flanked by 50 bp homology arms. Following microinjection, zygotes were exposed to 0, 3.75, or 7.5 µM RS‐1 for 24 hr. No differences were noted between groups in terms of development or genome edition rates. However, targeted KI rates were doubled in the group exposed to 7.5 µM RS‐1 compared to the others (52.8% vs. 25% and 23.1%, for 7.5, 0, and 3.75 µM, respectively). In conclusion, transient exposure to 7.5 µM RS‐1 enhances targeted KI rates resulting in approximately half of the embryos containing the intended mutation, hence allowing direct KI generation in embryos.
RS‐1 biases DNA repair towards homologous directed repair (HDR). Enhancing HDR increases the efficiency of clustered regularly interspaced short palindromic repeats (CRISPR)‐mediated targeted KI. Following transient RS‐1 exposure half of the injected bovine embryos contain the intended DNA insertion.
Summary Background There is evidence that meticillin-resistant Staphylococcus aureus (MRSA) bacteraemia can be reduced with improved infection control and antibiotic stewardship. Aim To survey ...infection control and antibiotic stewardship practices within European hospitals and to identify initiatives that correlate with reduced MRSA prevalence. Methods Online questionnaires were sent to European hospitals about their surveillance, hand hygiene, intravenous device management, admission screening, isolation, antibiotic prescribing, hospital demographics and MRSA blood culture isolates during 2010. Findings In all, 269 replies were received from hospitals in 29 European countries. Lower MRSA prevalence showed significant association with presence of incidence surveillance, performance of root cause analysis, mandatory training requirements for hand hygiene, accountability measures for persistent non-compliance, and multi-stakeholder teamwork in antibiotic prescribing. Presence of policies on intravenous catheter insertion and management showed no variation between different MRSA prevalence groups. However, low-prevalence hospitals reported more competency assessment programmes in insertion and maintenance of peripheral and central venous catheters. Hospitals from the UK and Ireland reported the highest uptake of infection control and antibiotic stewardship practices that were significantly associated with low MRSA prevalence, whereas Southern European hospitals exhibited the lowest. In multiple regression analysis, isolation of high-risk patients, performance of root cause analysis, obligatory training for nurses in hand hygiene, and undertaking joint ward rounds including microbiologists and infectious disease physicians remained significantly associated with lower MRSA prevalence. Conclusion Proactive infection control and antibiotic stewardship initiatives that instilled accountability, ownership, teamwork, and validated competence among healthcare workers were associated with improved MRSA outcomes.
Gaucher disease (GD) and Fabry disease (FD) are two relatively common inherited glycosphingolipidoses caused by deficiencies in the lysosomal glycosidases glucocerebrosidase and alpha-galactosidase ...A, respectively. For both diseases enzyme supplementation is presently used as therapy. Cells and tissues of GD and FD patients are uniformly deficient in enzyme activity, but the two diseases markedly differ in cell types showing lysosomal accumulation of the glycosphingolipid substrates glucosylceramide and globotriaosylceramide, respectively. The clinical manifestation of Gaucher disease and Fabry disease is consequently entirely different and the response to enzyme therapy is only impressive in the case of GD patients. This review compares both glycosphingolipid storage disorders with respect to similarities and differences. Presented is an update on insights regarding pathophysiological mechanisms as well as recently available biochemical markers and diagnostic tools for both disorders. Special attention is paid to sphingoid bases of the primary storage lipids in both diseases. The value of elevated glucosylsphingosine in Gaucher disease and globotriaosylsphingosine in Fabry disease for diagnosis and monitoring of disease is discussed as well as the possible contribution of the sphingoid bases to (patho)physiology. This article is part of a Special Issue entitled New Frontiers in Sphingolipid Biology.
•Gaucher and Fabry diseases are both lysosomal glycosphingolipid storage disorders due to glycosidase deficiencies.•Clinical manifestations, diagnostic issues, and therapy approaches for both diseases are viewed.•The occurrence and possible implications of lyso-glycosphingolipids in both diseases are discussed.•Design and use of activity-based probes allowing visualization of active glycosidases in situ are described.
Context.
Magnetic bright points (MBPs) are dynamic, small-scale magnetic elements often found with field strengths of the order of a kilogauss within intergranular lanes in the photosphere.
Aims.
...Here we study the evolution of various physical properties inferred from inverting high-resolution full Stokes spectropolarimetry data obtained from ground-based observations of the quiet Sun at disc centre.
Methods.
Using automated feature-tracking algorithms, we studied 300 MBPs and analysed their temporal evolution as they evolved to kilogauss field strengths. These properties were inferred using both the NICOLE and SIR Stokes inversion codes. We employ similar techniques to study radiative magnetohydrodynamical simulations for comparison with our observations.
Results.
Evidence was found for fast (∼30−100 s) amplification of magnetic field strength (by a factor of 2 on average) in MBPs during their evolution in our observations. Similar evidence for the amplification of fields is seen in our simulated data.
Conclusions.
Several reasons for the amplifications were established, namely, strong downflows preceding the amplification (convective collapse), compression due to granular expansion and mergers with neighbouring MBPs. Similar amplification of the fields and interpretations were found in our simulations, as well as amplification due to vorticity. Such a fast amplification will have implications for a wide array of topics related to small-scale fields in the lower atmosphere, particularly with regard to propagating wave phenomena in MBPs.
Anosognosia for hemiplegia (AHP) is informative about the neurocognitive basis of motor awareness. However, it is frequently associated with concomitant symptoms, such as hemispatial neglect and ...disturbances in the sense of body ownership (DSO). Although double dissociations between these symptoms have been reported, there is ongoing debate about whether they are manifestations of independent abnormalities, or a single neurocognitive deficit. We aimed to investigate the specificity of lesions associated with AHP by surpassing four, existing methodological limitations: (a) recruit a relatively large sample of patients (total N = 70) in a multi-centre study; (b) identify lesions associated with AHP in grey and white matter using voxel-based methods; (c) take into account the duration of AHP and concomitant neglect symptoms; and (d) compare lesions against a control hemiplegic group, patients suffering from AHP and DSO, and a few, rare patients with selective DSO. Results indicated that acute AHP is associated with a wide network, mainly including: (1) the Rolandic operculum, (2) the insula and (3) the superior temporal gyri. Subcortically, damage mainly involved the basal ganglia and white matter, mostly the superior corona radiate, arcuate fasciculus and the part of the ventral, superior longitudinal fasciculus. Persistent symptoms were linked with wider damage involving fronto-temporal cortex and long white matter tracts. A shift in the latero-medial direction (mainly involving the basal ganglia and surrounding white matter) emerged when DSO was taken accounted for. These results suggest that while bodily awareness is processed by areas widely distributed across the brain, intact subcortical structures and white matter tracts may be necessary to support basic feelings of owning and controlling contralateral body parts. An accurate and ‘up-to-date’ awareness of our motor abilities, however, may rely also on intact processing in cortical areas which presumably allow higher-order inferences about the current state of the body.
Objective
Brain structural changes in schizoaffective disorder, and how far they resemble those seen in schizophrenia and bipolar disorder, have only been studied to a limited extent.
Method
...Forty‐five patients meeting DSM‐IV and RDC criteria for schizoaffective disorder, groups of patients with 45 matched schizophrenia and bipolar disorder, and 45 matched healthy controls were examined using voxel‐based morphometry (VBM).
Results
Analyses comparing each patient group with the healthy control subjects found that the patients with schizoaffective disorder and the patients with schizophrenia showed widespread and overlapping areas of significant volume reduction, but the patients with bipolar disorder did not. A subsequent analysis compared the combined group of patients with the controls followed by extraction of clusters. In regions where the patients differed significantly from the controls, no significant differences in mean volume between patients with schizoaffective disorder and patients with schizophrenia in any of five regions of volume reduction were found, but mean volumes in the patients with bipolar disorder were significantly smaller in three of five.
Conclusion
The findings provide evidence that, in terms of structural gray matter brain abnormality, schizoaffective disorder resembles schizophrenia more than bipolar disorder.
The present work is the first of a two‐part paper on the Embrace Magnetometer Network. In this part, we present the new Embrace Magnetometer Network (Embrace MagNet) in South America, which is ...originally planned to cover most of the eastern portion of the Southern America longitudinal sector by installing and operating fluxgate magnetometer stations. We discuss the purpose and scientific goals of the network, associated with aeronomy and space weather. We provide details on the instrumentation, location of the sensors, sensitivity matching process, gain matching process, and magnetometer installation. In addition, we present and discuss details about the data storage, near‐real time display, and availability.
Plain Language Summary
This manuscript introduces the reader to the new Embrace Magnetometer Network (Embrace MagNet), which is located in South America and is based on fluxgate magnetometer. Its main purpose is to fulfill the gap in magnetic measurements, which are suitable for space weather purpose. Details on the instrumentation, location of the sensors, sensitivity matching process, gain matching process, and magnetometer installation are provided in this first of a two‐part paper on the Embrace MagNet. The accompanying paper provides information on the first scientific findings.
Key Points
Magnetic measurements in South America ready for space weather
Magnetic station measurement with near‐observatory data quality
New real‐time data providing proxy for Dst, and the new Ksa magnetic indices
ABSTRACT
We present NuSTAR (Nuclear Spectroscopic Telescope Array) observations of four active galactic nuclei (AGNs) located within 15 Mpc. These AGNs, namely ESO 121-G6, NGC 660, NGC 3486, and NGC ...5195, have observed X-ray luminosities of L2–10 keV,obs ≲ 1039 erg s−1, classifying them as low-luminosity AGN (LLAGN). We perform broad-band X-ray spectral analysis for the AGN by combining our NuSTAR data with Chandra or XMM–Newton observations to directly measure their column densities (NH) and infer their intrinsic power. We complement our X-ray data with archival and new high-angular resolution mid-infrared (mid-IR) data for all objects, except NGC 5195. Based on our X-ray spectral analysis, we found that both ESO 121-G6 and NGC 660 are heavily obscured (NH > 1023 cm−2; L2–10 keV,int ∼ 1041 erg s−1), and NGC 660 may be Compton thick. We also note that the X-ray flux and spectral slope for ESO 121-G6 have significantly changed over the last decade, indicating significant changes in the obscuration and potentially accretion rate. On the other hand, NGC 3486 and NGC 5195 appear to be unobscured and just mildly obscured, respectively, with L2–10 keV,int < 1039 erg s−1, i.e. genuine LLAGN. Both of the heavily obscured AGNs have Lbol > 1041 erg s−1 and λEdd ≳ 10−3, and are detected in high-angular resolution mid-IR imaging, indicating the presence of obscuring dust on nuclear scale. NGC 3486, however, is undetected in high-resolution mid-IR imaging, and the current data do not provide stringent constraints on the presence or absence of obscuring nuclear dust in the AGN.
We report observations from the Spitzer Space Telescope regarding the frequency of 24 mu m excess emission toward Sun-like stars. Our unbiased sample is composed of 309 stars with masses 0.7-2.2 M ...unk and ages from <3 Myr to >3 Gyr that lack excess emission at wavelengths less than or equal to 8 mu m. We identify 30 stars that exhibit clear evidence of excess emission from the observed 24 mu m/8 mu m flux ratio. The implied 24 mu m excesses of these candidate debris disk systems range from 13% (the minimum detectable) to more than 100% compared to the expected photospheric emission. The frequency of systems with evidence for dust debris emitting at 24 mu m ranges from 8.5%-19% at ages <300 Myr to <4% for older stars. The results suggest that many, perhaps most, Sun-like stars might form terrestrial planets.
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