The functional analysis of nonsense suppressors derived from in vitro engineered \u3ci\u3eSaccharomyces cerevisiae\u3c/i\u3e tRNA\u3csup\u3eTrp\u3c/sup\u3e genes

Nonsense suppressors derived from Saccharomyces cerevisiae tRNATrp genes have not been identified by classical genetic screens, although one can construct efficient amber (am) suppressors from them by making the appropriate anticodon mutation in vitro. Herein, a series of in vitro constructed putative suppressor genes was produced to test if pre-tRNATrp processing difficulties could help to explain the lack of classical tRNATrp-based suppressors. It is clear that inefficient processing of in-trons from precursor tRNATrp, or inaccurate overall processing, may explain why some of these constructs fail to promote nonsense suppression in vivo. However, deficient processing must be only one of the reasons why classical tRNATrp-based suppressors have not been characterized, as suppres-sion may still be extremely weak or absent in instances where the in vitro construct can lead to an accumulation of mature tRNATrp. Furthermore, suppression is also very weak in strains transformed with an intronless derivative of a putative tRNATrp ochre (oc) suppressor gene, wherein intron removal cannot pose a problem

The functional analysis of nonsense suppressors derived from in vitro engineered \u3ci\u3eSaccharomyces cerevisiae\u3c/i\u3e tRNA\u3csup\u3eTrp\u3c/sup\u3e genes

Nonsense suppressors derived from Saccharomyces cerevisiae tRNATrp genes have not been identified by classical genetic screens, although one can construct efficient amber (am) suppressors from them by making the appropriate anticodon mutation in vitro. Herein, a series of in vitro constructed putative suppressor genes was produced to test if pre-tRNATrp processing difficulties could help to explain the lack of classical tRNATrp-based suppressors. It is clear that inefficient processing of in-trons from precursor tRNATrp, or inaccurate overall processing, may explain why some of these constructs fail to promote nonsense suppression in vivo. However, deficient processing must be only one of the reasons why classical tRNATrp-based suppressors have not been characterized, as suppres-sion may still be extremely weak or absent in instances where the in vitro construct can lead to an accumulation of mature tRNATrp. Furthermore, suppression is also very weak in strains transformed with an intronless derivative of a putative tRNATrp ochre (oc) suppressor gene, wherein intron removal cannot pose a problem

The Local Group Census: planetary nebulae in Sextans B

Five planetary nebulae (PNe) have been discovered in the nearby dwarf irregular galaxy. Emission line images were obtained using the Wide Field Camera of the 2.5m Isaac Newton Telescope (INT) at La Palma (Spain). The candidate PNe were identified by their point-like appearance and relatively strong [OIII] emission-line fluxes. They are located within a galactocentric distance of 2.8 arcmin, corresponding to 1.1 kpc at the distance of Sextans B. Luminosities are in the range 1800--5600Lsolar. Sextans B is one of the smallest dwarf irregular galaxies with a PN population. The number of PNe detected suggest an enhanced star formation rate between 1 and 5 Gyr ago.Comment: 7 pages, 2 figure

Identifying Old Tidal Dwarf Irregulars

We examine the observational consequences of the two possible origins for irregular galaxies: formation from collapse of a primordial cloud of gas early in the age of the Universe, and formation from tidal tails in an interaction that could have occured any time in the history of the Universe. Because the formation from tidal tails could have occurred a long time ago, proximity to larger galaxies is not sufficient to distinguish tidal dwarfs from traditional dwarfs. We consider the effects of little or no dark matter on rotation speeds and the Tully-Fisher relationship, the metallicity-luminosity relationship, structure, and stellar populations. From these selection criteria, we identify a small list of dwarf irregular galaxies that are candidates for having formed as tidal dwarfs.Comment: ApJ, to appear September 20, 200

Optical Counterparts of Ultra-Luminous X-ray Sources identified from Archival Hubble Space Telescope/WFPC2

We present a systematic analysis of archival HST WFPC2 ``Association'' data sets that correlate with the Chandra positions of a set of 44 ultra-luminous X-ray sources (ULXs) of nearby galaxies. We have improved the Chandra-HST relative astrometry whenever possible. Disparate numbers of potential ULX counterparts are found, and in some cases none are found. The lack of or low number of counterparts in some cases may be due to insufficient depth in the WFPC2 images. Particularly in late-type galaxies, the HST image in the ULX region was often complex or crowded. We therefore address various scenarios for the nature of the ULX since it is not known which, if any, of the sources found are true counterparts. The optical luminosities of the sources are typically in the range 10^4-6 L_sun. In several cases color information is available, with the colors roughly tending to be more red in early-type galaxies. This suggests that, in general, the (potential) counterparts found in early-type galaxies are likely to be older stellar populations, and are probably globular clusters. Several early-type galaxy counterparts have blue colors, which may be due to younger stellar populations in the host galaxies, however these could also be background sources. In spiral galaxies the sources may also be due to localized structure in the disks rather than bound stellar systems. Alternatively some of the counterparts in late-type galaxies may be isolated supergiant stars. The observed X-ray/optical flux ratio is diluted by the optical emission of the cluster in cases where the system is an X-ray binary in a cluster, particularly in the case of a low-mass X-ray binaries in old cluster. (abridged)Comment: 35 pages with 9 figures formatted with emulateapj. Only subset of figures 1 and 2 are shown, for full version see http://xassist.pha.jhu.edu/ptak/hst_ulx_pape

Systematic review of antiepileptic drugs’ safety and effectiveness in feline epilepsy

Understanding the efficacy and safety profile of antiepileptic drugs (AEDs) in feline epilepsy is a crucial consideration for managing this important brain disease. However, there is a lack of information about the treatment of feline epilepsy and therefore a systematic review was constructed to assess current evidence for the AEDs’ efficacy and tolerability in cats. The methods and materials of our former systematic reviews in canine epilepsy were mostly mirrored for the current systematic review in cats. Databases of PubMed, CAB Direct and Google scholar were searched to detect peer-reviewed studies reporting efficacy and/or adverse effects of AEDs in cats. The studies were assessed with regards to their quality of evidence, i.e. study design, study population, diagnostic criteria and overall risk of bias and the outcome measures reported, i.e. prevalence and 95% confidence interval of the successful and affected population in each study and in total

Altered Gut Microbiota and Endocannabinoid System Tone in Obese and Diabetic Leptin-Resistant Mice: Impact on Apelin Regulation in Adipose Tissue

Growing evidence supports the role of gut microbiota in the development of obesity, type 2 diabetes, and low-grade inflammation. The endocrine activity of adipose tissue has been found to contribute to the regulation of glucose homeostasis and low-grade inflammation. Among the key hormones produced by this tissue, apelin has been shown to regulate glucose homeostasis. Recently, it has been proposed that gut microbiota participate in adipose tissue metabolism via the endocannabinoid system (eCB) and gut microbiota-derived compounds, namely lipopolysaccharide (LPS). We have investigated gut microbiota composition in obese and diabetic leptin-resistant mice (db/db) by combining pyrosequencing and phylogenetic microarray analysis of 16S ribosomal RNA gene sequences. We observed a significant higher abundance of Firmicutes, Proteobacteria, and Fibrobacteres phyla in db/db mice compared to lean mice. The abundance of 10 genera was significantly affected by the genotype. We identified the roles of the eCB and LPS in the regulation of apelinergic system tone (apelin and APJ mRNA expression) in genetic obese and diabetic mice. By using in vivo and in vitro models, we have demonstrated that both the eCB and low-grade inflammation differentially regulate apelin and APJ mRNA expression in adipose tissue. Finally, deep-gut microbiota profiling revealed that the gut microbial community of type 2 diabetic mice is significantly different from that of their lean counterparts. This indicates specific relationships between the gut microbiota and the regulation of the apelinergic system. However, the exact roles of specific bacteria in shaping the phenotype of db/db mice remain to be determined

Validity and responsiveness of the Clubfoot Assessment Protocol (CAP). A methodological study

BACKGROUND: The Clubfoot Assessment Protocol (CAP) is a multi dimensional instrument designed for longitudinal follow up of the clubfoot deformity during growth. Item reliability has shown to be sufficient. In this article the CAP's validity and responsiveness is studied using the Dimeglio classification scoring as a gold standard. METHODS: Thirty-two children with 45 congenital clubfeet were assessed prospectively and consecutively at ages of new-born, one, two, four months and two years of age. For convergent/divergent construct validity the Spearman's correlation coefficients were calculated. Discriminate validity was evaluated by studying the scores in bilateral clubfeet. The floor-ceiling effects at baseline (untreated clubfeet) and at two years of age (treated clubfeet) were evaluated. Responsiveness was evaluated by using effect sizes (ES) and by calculating if significant changes (Wilcoxons signed test) had occurred between the different measurement occasions. RESULTS: High to moderate significant correlation were found between CAP mobility I and morphology and the Dimeglio scores (r(s )= 0.77 and 0.44 respectively). Low correlation was found between CAP muscle function, mobility II and motion quality and the Dimeglio scoring system (r(s )= 0.20, 0.09 and 0.06 respectively). Of 13 children with bilateral clubfeet, 11 showed different CAP mobility I scores between right and left foot at baseline (untreated) compared with 5 with the Dimeglio score. At the other assessment occasions the CAP mobility I continued to show higher discrimination ability than the Dimeglio. No floor effects and low ceiling effects were found in the untreated clubfeet for both instruments. High ceiling effects were found in the CAP for the treated children and low for the Dimeglio. Responsiveness was good. ES from untreated to treated ranged from 0.80 to 4.35 for the CAP subgroups and was 4.68 for the Dimeglio. The first four treatment months, the CAP mobility I had generally higher ES compared with the Dimeglio. CONCLUSION: The Clubfoot Assessment Protocol shows in this study good validity and responsiveness. The CAP is more responsive when severity ranges between mild – moderate to severe, while the Dimeglio focuses more on the extremes. The ability to discriminate between different mobility status of the right and left foot in bilaterally affected children in this population was higher compared with the Dimeglio score implicating a better sensitivity for the CAP

Comparison of weather station and climate reanalysis data for modelling temperature-related mortality

Epidemiological analyses of health risks associated with non-optimal temperature are traditionally based on ground observations from weather stations that offer limited spatial and temporal coverage. Climate reanalysis represents an alternative option that provide complete spatio-temporal exposure coverage, and yet are to be systematically explored for their suitability in assessing temperature-related health risks at a global scale. Here we provide the first comprehensive analysis over multiple regions to assess the suitability of the most recent generation of reanalysis datasets for health impact assessments and evaluate their comparative performance against traditional station-based data. Our findings show that reanalysis temperature from the last ERA5 products generally compare well to station observations, with similar non-optimal temperature-related risk estimates. However, the analysis offers some indication of lower performance in tropical regions, with a likely underestimation of heat-related excess mortality. Reanalysis data represent a valid alternative source of exposure variables in epidemiological analyses of temperature-related risk