Synthetic long non-coding RNAs [SINEUPs] rescue defective gene expression in vivo

Non-coding RNAs provide additional regulatory layers to gene expression as well as the potential to being exploited as therapeutic tools. Non-coding RNA-based therapeutic approaches have been attempted in dominant diseases, however their use for treatment of genetic diseases caused by insufficient gene dosage is currently more challenging. SINEUPs are long antisense non-coding RNAs that up-regulate translation in mammalian cells in a gene-specific manner, although, so far evidence of SINEUP efficacy has only been demonstrated in in vitro systems. We now show that synthetic SINEUPs effectively and specifically increase protein levels of a gene of interest in vivo. We demonstrated that SINEUPs rescue haploinsufficient gene dosage in a medakafish model of a human disorder leading to amelioration of the disease phenotype. Our results demonstrate that SINEUPs act through mechanisms conserved among vertebrates and that SINEUP technology can be successfully applied in vivo as a new research and therapeutic tool for gene-specific up-regulation of endogenous functional proteins

V5852 Sgr : an unusual nova possibly associated with the Sagittarius stream

We report spectroscopic and photometric follow-up of the peculiar nova V5852~Sgr (discovered as OGLE-2015-NOVA-01), which exhibits a combination of features from different nova classes. The photometry shows a flat-topped light curve with quasi-periodic oscillations, then a smooth decline followed by two fainter recoveries in brightness. Spectroscopy with the Southern African Large Telescope shows first a classical nova with an Fe II or Fe IIb spectral type. In the later spectrum, broad emissions from helium, nitrogen and oxygen are prominent and the iron has faded which could be an indication to the start of the nebular phase. The line widths suggest ejection velocities around 1000 km s-1. The nova is in the direction of the Galactic bulge and is heavily reddened by an uncertain amount. The V magnitude 16 days after maximum enables a distance to be estimated and this suggests that the nova may be in the extreme trailing stream of the Sagittarius dwarf spheroidal galaxy. If so it is the first nova to be detected from that, or from any dwarf spheroidal galaxy. Given the uncertainty of the method and the unusual light curve we cannot rule out the possibility that it is in the bulge or even the Galactic disk behind the bulge.Publisher PDFPeer reviewe

Highly Asynchronous and Asymmetric Cleavage Divisions Accompany Early Transcriptional Activity in Pre-Blastula Medaka Embryos

In the initial phase of development of fish embryos, a prominent and critical event is the midblastula transition (MBT). Before MBT cell cycle is rapid, highly synchronous and zygotic gene transcription is turned off. Only during MBT the cell cycle desynchronizes and transcription is activated. Multiple mechanisms, primarily the nucleocytoplasmic ratio, are supposed to control MBT activation. Unexpectedly, we find in the small teleost fish medaka (Oryzias latipes) that at very early stages, well before midblastula, cell division becomes asynchronous and cell volumes diverge. Furthermore, zygotic transcription is extensively activated already after the 64-cell stage. Thus, at least in medaka, the transition from maternal to zygotic transcription is uncoupled from the midblastula stage and not solely controlled by the nucleocytoplasmic ratio

Gene expression throughout a vertebrate's embryogenesis

Abstract Background Describing the patterns of gene expression during embryonic development has broadened our understanding of the processes and patterns that define morphogenesis. Yet gene expression patterns have not been described throughout vertebrate embryogenesis. This study presents statistical analyses of gene expression during all 40 developmental stages in the teleost Fundulus heteroclitus using four biological replicates per stage. Results Patterns of gene expression for 7,000 genes appear to be important as they recapitulate developmental timing. Among the 45% of genes with significant expression differences between pairs of temporally adjacent stages, significant differences in gene expression vary from as few as five to more than 660. Five adjacent stages have disproportionately more significant changes in gene expression (> 200 genes) relative to other stages: four to eight and eight to sixteen cell stages, onset of circulation, pre and post-hatch, and during complete yolk absorption. The fewest differences among adjacent stages occur during gastrulation. Yet, at stage 16, (pre-mid-gastrulation) the largest number of genes has peak expression. This stage has an over representation of genes in oxidative respiration and protein expression (ribosomes, translational genes and proteases). Unexpectedly, among all ribosomal genes, both strong positive and negative correlations occur. Similar correlated patterns of expression occur among all significant genes. Conclusions These data provide statistical support for the temporal dynamics of developmental gene expression during all stages of vertebrate development

Structural Conformers of (1,3-Dithiol-2-ylidene)ethanethioamides: The Balance Between Thioamide Rotation and Preservation of Classical Sulfur-Sulfur Hypervalent Bonds

The reaction of N-(2-phthalimidoethyl)-N-alkylisopropylamines and S2Cl2 gave 4-N-(2-phthalimidoethyl)-N-alkylamino-5-chloro-1,2-dithiol-3-thiones that quantitatively cycloadded to dimethyl or diethyl acetylenedicarboxylate to give stable thioacid chlorides, which in turn reacted with one equivalent of aniline or a thiole to give thioanilides or a dithioester. Several compounds of this series showed atropisomers that were studied by a combination of dynamic NMR, simulation of the signals, conformational analysis by DFT methods, and single crystal X-ray diffraction, showing a good correlation between the theoretical calculations, the experimental values of energies, and the preferred conformations in the solid state. The steric hindering of the crowded substitution at the central amine group was found to be the reason for the presence of permanent atropisomers in this series of compounds and the cause of a unique disposition of the thioxo group at close-to-right angles with respect to the plane defined by the 1,3-dithiole ring in the dithiafulvene derivatives, thus breaking the sulfur–sulfur hypervalent bond that is always found in this kind of compounds.Ministerio de Economıá y Competitividad, Spain (Project CTQ2012- 31611), Junta de Castilla y León, Consejería de Educación y Cultura y Fondo Social Europeo (Project BU246A12-1), and the European Commission, Seventh Framework Programme (Project SNIFFER FP7-SEC-2012-312411

Analysis of the Ush2a Gene in Medaka Fish (Oryzias latipes)

Patients suffering from Usher syndrome (USH) exhibit sensorineural hearing loss, retinitis pigmentosa (RP) and, in some cases, vestibular dysfunction. USH is the most common genetic disorder affecting hearing and vision and is included in a group of hereditary pathologies associated with defects in ciliary function known as ciliopathies. This syndrome is clinically classified into three types: USH1, USH2 and USH3. USH2 accounts for well over one-half of all Usher cases and mutations in the USH2A gene are responsible for the majority of USH2 cases, but also for atypical Usher syndrome and recessive non-syndromic RP. Because medaka fish (Oryzias latypes) is an attractive model organism for genetic-based studies in biomedical research, we investigated the expression and function of the USH2A ortholog in this teleost species. Ol-Ush2a encodes a protein of 5.445 aa codons, containing the same motif arrangement as the human USH2A. Ol-Ush2a is expressed during early stages of medaka fish development and persists into adulthood. Temporal Ol-Ush2a expression analysis using whole mount in situ hybridization (WMISH) on embryos at different embryonic stages showed restricted expression to otoliths and retina, suggesting that Ol-Ush2a might play a conserved role in the development and/or maintenance of retinal photoreceptors and cochlear hair cells. Knockdown of Ol-Ush2a in medaka fish caused embryonic developmental defects (small eyes and heads, otolith malformations and shortened bodies with curved tails) resulting in late embryo lethality. These embryonic defects, observed in our study and in other ciliary disorders, are associated with defective cell movement specifically implicated in left-right (LR) axis determination and planar cell polarity (PCP)

The WEBT BL Lac Campaign 2000

We present UBVRI light curves of BL Lacertae from May 2000 to January 2001, obtained by 24 telescopes in 11 countries. More than 15000 observations were performed in that period, which was the extension of the Whole Earth Blazar Telescope (WEBT) campaign originally planned for July-August 2000. Rapid flux oscillations are present all the time, involving variations up to a few tenths of mag on hour time scales, and witnessing an intense intraday activity of this source. Colour indexes have been derived by coupling the highest precision B and R data taken by the same instrument within 20 min and after subtracting the host galaxy contribution from the fluxes. The 620 indexes obtained show that the optical spectrum is weakly sensitive to the long-term trend, while it strictly follows the short-term flux behaviour, becoming bluer when the brightness increases. Thus, spectral changes are not related to the host galaxy contribution, but they are an intrinsic feature of fast flares. We suggest that the achromatic mechanism causing the long-term flux base-level modulation can be envisaged in a variation of the relativistic Doppler beaming factor, and that this variation is likely due to a change of the viewing angle. Discrete correlation function (DCF) analysis reveals the existence of a characteristic time scale of variability of about 7 h in the light curve of the core WEBT campaign, while no measurable time delay between variations in the B and R bands is found.Comment: 14 pages, 8 PostScript figures, 5 JPEG figures, in press for A&

Coordinated Multiwavelength Observations of BL Lacertae in 2000

BL Lacertae was the target of an extensive multiwavelength monitoring campaign in the second half of 2000. Simultaneous or quasi-simultaneous observations were taken at radio (UMRAO and Metsaehovi) and optical(WEBT collaboration) frequencies, in X-rays (BeppoSAX and RXTE), and at VHE gamma-rays (HEGRA). The WEBT optical campaign achieved an unprecedented time coverage, virtually continuous over several 10 - 20 hour segments. It revealed intraday variability on time scales of ~ 1.5 hours and evidence for spectral hardening associated with increasing optical flux. During the campaign, BL Lacertae underwent a major transition from a rather quiescent state prior to September 2000, to a flaring state for the rest of the year. This was also evident in the X-ray activity of the source. BeppoSAX observations on July 26/27 revealed a rather low X-ray flux and a hard spectrum, while a BeppoSAX pointing on Oct. 31 - Nov. 2, 2000, indicated significant variability on time scales of < a few hours, and provided evidence for the synchrotron spectrum extending out to ~ 10 keV during that time. During the July 26/27 observation, there is a tantalizing, though not statistically significant, indication of a time delay of ~ 4 - 5 hr between the BeppoSAX and the R-band light curve. Also, a low-significance detection of a time delay of 15 d between the 14.5 GHz and the 22 GHz radio light curves is reported. Several independent methods to estimate the co-moving magnetic field in the source are presented, suggesting a value of ~ 2 e_B^{2/7} G, where e_B is the magnetic-field equipartition factor w.r.t. the electron energy density in the jet.Comment: Accepted for publication in Ap

Calcium ion currents mediating oocyte maturation events

During maturation, the last phase of oogenesis, the oocyte undergoes several changes which prepare it to be ovulated and fertilized. Immature oocytes are arrested in the first meiotic process prophase, that is morphologically identified by a germinal vesicle. The removal of the first meiotic block marks the initiation of maturation. Although a large number of molecules are involved in complex sequences of events, there is evidence that a calcium increase plays a pivotal role in meiosis re-initiation. It is well established that, during this process, calcium is released from the intracellular stores, whereas less is known on the role of external calcium entering the cell through the plasma membrane ion channels. This review is focused on the functional role of calcium currents during oocyte maturation in all the species, from invertebrates to mammals. The emerging role of specific L-type calcium channels will be discussed