Gene expression of O-GlcNAc cycling enzymes in human breast cancers

O-GlcNAcylation is an abundant, dynamic, and inducible posttranslational modification in which single β-N-acetylglucosamine residues are attached by O-glycosidic linkage to serine or treonine residues. It is suggested that abnormally regulated O-GlcNAcylation may contribute to the pathology of cancer. Cycling of O-GlcNAc residues on intracellular proteins is controlled by two enzymes, O-GlcNAc transferease (OGT), which catalyses the addition of O-GlcNAc residues and nucleocytoplasmic β-N-acetylglucosaminidase (O-GlcNAcase; encoded by MGEA5 gene), an enzyme involved in the removal of O-GlcNAc. In this study, relationship between the mRNA expressions of genes coding O-GlcNAc cycling enzymes in breast ductal carcinomas and clinicopathological parameters were analyzed. The results showed that poorly differentiated tumors (grade II and III) had significantly higher OGT expression than grade I tumors. Contrary, MGEA5 transcript levels were significantly lower in grade II and III in comparison with grade I tumors. The Spearman rank correlation showed the expressions of OGT and MGEA5 in breast cancer was negatively correlated (r = −0.430, P = 0.0002). Lymph node metastasis status was significantly associated with decreased MGEA5 mRNA expression. This result suggests that elevation in O-GlcNAc modification of proteins may be implicated in breast tumor progression and metastasis

Regulation of Dopamine Release by CASK-β Modulates Locomotor Initiation in Drosophila melanogaster

CASK is an evolutionarily conserved scaffolding protein that has roles in many cell types. In Drosophila, loss of the entire CASK gene or just the CASK- transcript causes a complex set of adult locomotor defects. In this study, we show that the motor initiation component of this phenotype is due to loss of CASK- in dopaminergic neurons and can be specifically rescued by expression of CASK- within this subset of neurons. Functional imaging demonstrates that mutation of CASK- disrupts coupling of neuronal activity to vesicle fusion. Consistent with this, locomotor initiation can be rescued by artificially driving activity in dopaminergic neurons. The molecular mechanism underlying this role of CASK- in dopaminergic neurons involves interaction with Hsc70-4, a molecular chaperone previously shown to regulate calcium-dependent vesicle fusion. These data suggest that there is a novel CASK- -dependent regulatory complex in dopaminergic neurons that serves to link activity and neurotransmitter release.Fil: Slawson, Justin B. Brandeis University; Estados UnidosFil: Kuklin, Elena A. Brandeis University; Estados UnidosFil: Mukherjee, Konark. Brandeis University; Estados UnidosFil: Pírez, Nicolas. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquimicas de Buenos Aires; Argentina. Fundación Instituto Leloir; Argentina. Brandeis University; Estados UnidosFil: Donelson, Nathan C. Brandeis University; Estados UnidosFil: Griffith, Leslie C. Brandeis University; Estados Unido

Constraining the False Positive Rate for Kepler Planet Candidates with Multi-Color Photometry from the GTC

Using the OSIRIS instrument installed on the 10.4-m Gran Telescopio Canarias (GTC) we acquired multi-color transit photometry of four small (Rp < 5 R_Earth) short-period (P < 6 days) planet candidates recently identified by the Kepler space mission. These observations are part of a program to constrain the false positive rate for small, short-period Kepler planet candidates. Since planetary transits should be largely achromatic when observed at different wavelengths (excluding the small color changes due to stellar limb darkening), we use the observed transit color to identify candidates as either false positives (e.g., a blend with a stellar eclipsing binary either in the background/foreground or bound to the target star) or validated planets. Our results include the identification of KOI 225.01 and KOI 1187.01 as false positives and the tentative validation of KOI 420.01 and KOI 526.01 as planets. The probability of identifying two false positives out of a sample of four targets is less than 1%, assuming an overall false positive rate for Kepler planet candidates of 10% (as estimated by Morton & Johnson 2011). Therefore, these results suggest a higher false positive rate for the small, short-period Kepler planet candidates than has been theoretically predicted by other studies which consider the Kepler planet candidate sample as a whole. Furthermore, our results are consistent with a recent Doppler study of short-period giant Kepler planet candidates (Santerne et al. 2012). We also investigate how the false positive rate for our sample varies with different planetary and stellar properties. Our results suggest that the false positive rate varies significantly with orbital period and is largest at the shortest orbital periods (P < 3 days), where there is a corresponding rise in the number of detached eclipsing binary stars... (truncated)Comment: 13 pages, 12 figures, 3 tables; revised for MNRA

A Process Description of Playing to Live! A Community Psychosocial Arts Program During Ebola

From 2014 to 2015, Liberia experienced the largest Ebola epidemic in world history. The impact of this disease was not only physical; it created fear, loss, and trauma throughout the country. This article will describe the process of three phases of a community-based psychosocial expressive arts program, which used theory from the fields of expressive arts therapy to build mental health capacity during and after the epidemic. This article will highlight the background of Ebola virus disease and the Ebola virus disease epidemic, provide an overview of current theory and research for expressive arts therapy and the impact of trauma, describe the process of how the program developed and was implemented, the process of partnering with the community, program components, the two pilot programs, and the large-scale community program. We performed a mixed-methods analysis of the large-scale program’s activity data to evaluate the impact. The results highlight a positive response from the participating children and facilitators. The authors discuss the findings from the results, best practices, and limitations. Additionally, the authors discuss implications and considerations for future programming

Stable transformation of an episomal protein-tagging shuttle vector in the piscine diplomonad Spironucleus vortens

<p>Abstract</p> <p>Background</p> <p>Diplomonads are common free-living inhabitants of anoxic aquatic environments and are also found as intestinal commensals or parasites of a wide variety of animals. <it>Spironucleus vortens </it>is a putatively commensal diplomonad of angelfish that grows to high cell densities in axenic culture. Genomic sequencing of <it>S. vortens </it>is in progress, yet little information is available regarding molecular and cellular aspects of <it>S. vortens </it>biology beyond descriptive ultrastructural studies. To facilitate the development of <it>S. vortens </it>as an additional diplomonad experimental model, we have constructed and stably transformed an episomal plasmid containing an enhanced green fluorescent protein (GFP) tag, an AU1 epitope tag, and a tandem affinity purification (TAP) tag. This construct also contains selectable antibiotic resistance markers for both <it>S. vortens </it>and <it>E. coli</it>.</p> <p>Results</p> <p>Stable transformants of <it>S. vortens </it>grew relatively rapidly (within 7 days) after electroporation and were maintained under puromycin selection for over 6 months. We expressed the enhanced GFP variant, eGFP, under transcriptional control of the <it>S. vortens </it>histone H3 promoter, and visually confirmed diffuse GFP expression in over 50% of transformants. Next, we generated a histone H3::GFP fusion using the <it>S. vortens </it>conventional histone H3 gene and its native promoter. This construct was also highly expressed in the majority of <it>S. vortens </it>transformants, in which the H3::GFP fusion localized to the chromatin in both nuclei. Finally, we used fluorescence <it>in situ </it>hybridization (FISH) of the episomal plasmid to show that the transformed plasmid localized to only one nucleus/cell and was present at roughly 10–20 copies per nucleus. Because <it>S. vortens </it>grows to high densities in laboratory culture, it is a feasible diplomonad from which to purify native protein complexes. Thus, we also included a TAP tag in the plasmid constructs to permit future tagging and subsequent purification of protein complexes by affinity chromatography via a two-step purification procedure.</p> <p>Conclusion</p> <p>Currently, progress in protistan functional and comparative genomics is hampered by the lack of free-living or commensal protists in axenic culture, as well as a lack of molecular genetic tools with which to study protein function in these organisms. This stable transformation protocol combined with the forthcoming genome sequence allows <it>Spironucleus vortens </it>to serve as a new experimental model for cell biological studies and for comparatively assessing protein functions in related diplomonads such as the human intestinal parasite, <it>Giardia intestinalis</it>.</p

A transiting companion to the eclipsing binary KIC002856960

We present an early result from an automated search of Kepler eclipsing binary systems for circumbinary companions. An intriguing tertiary signal has been discovered in the short period eclipsing binary KIC002856960. This third body leads to transit-like features in the light curve occurring every 204.2 days, while the two other components of the system display eclipses on a 6.2 hour period. The variations due to the tertiary body last for a duration of \sim1.26 days, or 4.9 binary orbital periods. During each crossing of the binary orbit with the tertiary body, multiple individual transits are observed as the close binary stars repeatedly move in and out of alignment with the tertiary object. We are at this stage unable to distinguish between a planetary companion to a close eclipsing binary, or a hierarchical triply eclipsing system of three stars. Both possibilities are explored, and the light curves presented.Comment: Accepted into A&A Letters (5 pages & 3 figures

OGA heterozygosity suppresses intestinal tumorigenesis in Apc min/+ mice

Emerging evidence suggests that aberrant O-GlcNAcylation is associated with tumorigenesis. Many oncogenic factors are O-GlcNAcylated, which modulates their functions. However, it remains unclear how O-GlcNAcylation and O-GlcNAc cycling enzymes, O-GlcNAc transferase (OGT) and O-GlcNAcase (OGA), affect the development of cancer in animal models. In this study, we show that reduced level of OGA attenuates colorectal tumorigenesis induced by Adenomatous polyposis coli (Apc) mutation. The levels of O-GlcNAcylation and O-GlcNAc cycling enzymes were simultaneously upregulated in intestinal adenomas from mice, and in human patients. In two independent microarray data sets, the expression of OGA and OGT was significantly associated with poor cancer-specific survival of colorectal cancer (CRC) patients. In addition, OGA heterozygosity, which results in increased levels of O-GlcNAcylation, attenuated intestinal tumor formation in the Apc min/+ background. Apc min/+ OGA +/-mice exhibited a significantly increased survival rate compared with Apc min/+ mice. Consistent with this, Apc min/+ OGA +/-mice expressed lower levels of Wnt target genes than Apc min/+. However, the knockout of OGA did not affect Wnt/??-catenin signaling. Overall, these findings suggest that OGA is crucial for tumor growth in CRC independently of Wnt/??-catenin signaling.open2

Call to adopt a nominal set of astrophysical parameters and constants to improve the accuracy of fundamental physical properties of stars

The increasing precision of astronomical observations of stars and stellar systems is gradually getting to a level where the use of slightly different values of the solar mass, radius and luminosity, as well as different values of fundamental physical constants, can lead to measurable systematic differences in the determination of basic physical properties. An equivalent issue with an inconsistent value of the speed of light was resolved by adopting a nominal value that is constant and has no error associated with it. Analogously, we suggest that the systematic error in stellar parameters may be eliminated by: (1) replacing the solar radius Rsun and luminosity Lsun by the nominal values that are by definition exact and expressed in SI units: 1 RnomSun = 6.95508 x 10^8 m and 1 LnomSun = 3.846 x 10^{26} W; (2) computing stellar masses in terms of Msun by noting that the measurement error of the product G.Msun is 5 orders of magnitude smaller than the error in G; (3) computing stellar masses and temperatures in SI units by using the derived values Msun(2010) = 1.988547 x 10^{30} kg and Tsun(2010) = 5779.57 K; and (4) clearly stating the reference for the values of the fundamental physical constants used. We discuss the need and demonstrate the advantages of such a paradigm shift.Comment: 6 pages, 3 table