The Role of miRNAs, miRNA Clusters, and isomiRs in Development of Cancer Stem Cell Populations in Colorectal Cancer.

MicroRNAs (miRNAs or miRs) have a critical role in regulating stem cells (SCs) duringdevelopment and altered expression can cause developmental defects and/or disease. Indeed,aberrant miRNA expression leads to wide-spread transcriptional dysregulation which has beenlinked to many cancers. Mounting evidence also indicates a role for miRNAs in the developmentof the cancer SC (CSC) phenotype. Our goal herein is to provide a review of: (i) current researchon miRNAs and their targets in colorectal cancer (CRC), and (ii) miRNAs that are differentiallyexpressed in colon CSCs. MicroRNAs can work in clusters or alone when targeting different SC genesto influence CSC phenotype. Accordingly, we discuss the specific miRNA cluster classifications andisomiRs that are predicted to target theALDH1,CD166,BMI1,LRIG1, andLGR5SC genes.miR-23bandmiR-92Aare of particular interest because our previously reported studies on miRNA expressionin isolated normal versus malignant human colonic SCs showed thatmiR-23bandmiR-92aareregulators of theLGR5andLRIG1SC genes, respectively. We also identify additional miRNAs whoseexpression inversely correlated with mRNA levels of their target genes and associated with CRCpatient survival. Altogether, our deliberation on miRNAs, their clusters, and isomiRs in regulationof SC genes could provide insight into how dysregulation of miRNAs leads to the emergence ofdifferent CSC populations and SC overpopulation in CRC

An exponential decline at the bright end of the z=6 galaxy luminosity function

We present the results of a search for the most luminous star-forming galaxies at redshifts z~6 based on CFHT Legacy Survey data. We identify a sample of 40 Lyman break galaxies brighter than magnitude z'=25.3 across an area of almost 4 square degrees. Sensitive spectroscopic observations of seven galaxies provide redshifts for four, of which only two have moderate to strong Lyman alpha emission lines. All four have clear continuum breaks in their spectra. Approximately half of the Lyman break galaxies are spatially resolved in 0.7 arcsec seeing images, indicating larger sizes than lower luminosity galaxies discovered with the Hubble Space Telescope, possibly due to on-going mergers. The stacked optical and infrared photometry is consistent with a galaxy model with stellar mass ~ 10^{10} solar masses. There is strong evidence for substantial dust reddening with a best-fit A_V=0.7 and A_V>0.48 at 2 sigma confidence, in contrast to the typical dust-free galaxies of lower luminosity at this epoch. The spatial extent and spectral energy distribution suggest that the most luminous z~6 galaxies are undergoing merger-induced starbursts. The luminosity function of z=5.9 star-forming galaxies is derived. This agrees well with previous work and shows strong evidence for an exponential decline at the bright end, indicating that the feedback processes which govern the shape of the bright end are occurring effectively at this epoch.Comment: 14 pages, 11 figures, AJ in press, revised to address referee comment

A robust system for RNA interference in the chicken using a modified microRNA operon

AbstractRNA interference (RNAi) provides an effective method to silence gene expression and investigate gene function. However, RNAi tools for the chicken embryo have largely been adapted from vectors designed for mammalian cells. Here we present plasmid and retroviral RNAi vectors specifically designed for optimal gene silencing in chicken cells. The vectors use a chicken U6 promoter to express RNAs modelled on microRNA30, which are embedded within chicken microRNA operon sequences to ensure optimal Drosha and Dicer processing of transcripts. The chicken U6 promoter works significantly better than promoters of mammalian origin and in combination with a microRNA operon expression cassette (MOEC), achieves up to 90% silencing of target genes. By using a MOEC, we show that it is also possible to simultaneously silence two genes with a single vector. The vectors express either RFP or GFP markers, allowing simple in vivo tracking of vector delivery. Using these plasmids, we demonstrate effective silencing of Pax3, Pax6, Nkx2.1, Nkx2.2, Notch1 and Shh in discrete regions of the chicken embryonic nervous system. The efficiency and ease of use of this RNAi system paves the way for large-scale genetic screens in the chicken embryo

RELICS: High-Resolution Constraints on the Inner Mass Distribution of the z=0.83 Merging Cluster RXJ0152.7-1357 from strong lensing

Strong gravitational lensing (SL) is a powerful means to map the distribution of dark matter. In this work, we perform a SL analysis of the prominent X-ray cluster RXJ0152.7-1357 (z=0.83, also known as CL 0152.7-1357) in \textit{Hubble Space Telescope} images, taken in the framework of the Reionization Lensing Cluster Survey (RELICS). On top of a previously known $z=3.93$ galaxy multiply imaged by RXJ0152.7-1357, for which we identify an additional multiple image, guided by a light-traces-mass approach we identify seven new sets of multiply imaged background sources lensed by this cluster, spanning the redshift range [1.79-3.93]. A total of 25 multiple images are seen over a small area of ~0.4 $arcmin^2$, allowing us to put relatively high-resolution constraints on the inner matter distribution. Although modestly massive, the high degree of substructure together with its very elongated shape make RXJ0152.7-1357 a very efficient lens for its size. This cluster also comprises the third-largest sample of z~6-7 candidates in the RELICS survey. Finally, we present a comparison of our resulting mass distribution and magnification estimates with those from a Lenstool model. These models are made publicly available through the MAST archive.Comment: 15 Pages, 7 Figures, 4 Tables Accepted for publication in Ap

Drug Susceptibility in Leishmania Isolates Following Miltefosine Treatment in Cases of Visceral Leishmaniasis and Post Kala-Azar Dermal Leishmaniasis

Resistance to antimonials has emerged as a major hurdle to the treatment and control of VL and led to the introduction of Miltefosine as first line treatment in the Indian subcontinent. MIL is an oral drug with a long half-life, and it is feared that resistance may emerge rapidly, threatening control efforts under the VL elimination program. There is an urgent need for monitoring treatment efficacy and emergence of drug resistance in the field. In a set of VL/PKDL cases recruited for MIL treatment, we observed comparable drug susceptibility in pre- and post-treatment isolates from cured VL patients while MIL susceptibility was significantly reduced in isolates from VL relapse and PKDL cases. The PKDL isolates showed higher tolerance to MIL as compared to VL isolates. Both VL and PKDL isolates were uniformly susceptible to PMM. MIL transporter genes LdMT/LdRos3 were previously reported as potential resistance markers in strains in which MIL resistance was experimentally induced. The point mutations and the down-regulated expression of these transporters observed in vitro could, however, not be verified in natural populations of parasites. LdMT/LdRos3 genes therefore, do not appear to be suitable markers so far for monitoring drug susceptibility in clinical leishmanial isolates

RELICS: Properties of z>5.5 Galaxies Inferred from Spitzer and Hubble Imaging Including A Candidate z~6.8 Strong [OIII] Emitter

We present constraints on the physical properties (including stellar mass, age, and star formation rate) of 207 $6\lesssim z \lesssim8$ galaxy candidates from the Reionization Lensing Cluster Survey (RELICS) and companion Spitzer-RELICS surveys. We measure photometry using T-PHOT and perform spectral energy distribution fitting using EA$z$Y and BAGPIPES. Of the 207 candidates for which we could successfully measure Spitzer fluxes, 23 were demoted to likely low redshift ($z<4$). Among the remaining high redshift candidates, we find intrinsic stellar masses between $1\times10^6\rm{M_{\odot}}$ and $4\times10^9\rm{M_\odot}$, and rest-frame UV absolute magnitudes between $-22.6$ and $-14.5$ mag. While our sample is mostly comprised of $L_{UV}/L^*_{UV}<1$ galaxies, there are a number of brighter objects in the sample, extending to $L_{UV}/L^*_{UV}\sim2$. The galaxies in our sample span approximately four orders of magnitude in stellar mass and star-formation rates, and exhibit ages ranging from maximally young to maximally old. We highlight 11 galaxies which have detections in Spitzer/IRAC imaging and redshift estimates $z\geq6.5$, several of which show evidence for some combination of evolved stellar populations, large contributions of nebular emission lines, and/or dust. Among these is PLCKG287+32-2013, one of the brightest $z\sim7$ candidates known (AB mag 24.9) with a Spitzer 3.6$\mu$m flux excess suggesting strong [OIII] + H-$\beta$ emission ($\sim$1000\AA\ rest-frame equivalent width). We discuss the possible uses and limits of our sample and present a public catalog of Hubble 0.4--1.6$\mu$m + Spitzer 3.6$\mu$m and 4.5$\mu$m photometry along with physical property estimates for all 207 objects in the sample. Because of their apparent brightnesses, high redshifts, and variety of stellar populations, these objects are excellent targets for follow-up with James Webb Space Telescope.Comment: 20 pages, 9 figure

Virtual Northern Analysis of the Human Genome

BACKGROUND: We applied the Virtual Northern technique to human brain mRNA to systematically measure human mRNA transcript lengths on a genome-wide scale. METHODOLOGY/PRINCIPAL FINDINGS: We used separation by gel electrophoresis followed by hybridization to cDNA microarrays to measure 8,774 mRNA transcript lengths representing at least 6,238 genes at high (>90%) confidence. By comparing these transcript lengths to the Refseq and H-Invitational full-length cDNA databases, we found that nearly half of our measurements appeared to represent novel transcript variants. Comparison of length measurements determined by hybridization to different cDNAs derived from the same gene identified clones that potentially correspond to alternative transcript variants. We observed a close linear relationship between ORF and mRNA lengths in human mRNAs, identical in form to the relationship we had previously identified in yeast. Some functional classes of protein are encoded by mRNAs whose untranslated regions (UTRs) tend to be longer or shorter than average; these functional classes were similar in both human and yeast. CONCLUSIONS/SIGNIFICANCE: Human transcript diversity is extensive and largely unannotated. Our length dataset can be used as a new criterion for judging the completeness of cDNAs and annotating mRNA sequences. Similar relationships between the lengths of the UTRs in human and yeast mRNAs and the functions of the proteins they encode suggest that UTR sequences serve an important regulatory role among eukaryotes

A Candidate z∼10z\sim10 Galaxy Strongly Lensed into a Spatially Resolved Arc

The most distant galaxies known are at z~10-11, observed 400-500 Myr after the Big Bang. The few z~10-11 candidates discovered to date have been exceptionally small- barely resolved, if at all, by the Hubble Space Telescope. Here we present the discovery of SPT0615-JD, a fortuitous z~10 (z_phot=9.9+/-0.6) galaxy candidate stretched into an arc over ~2.5" by the effects of strong gravitational lensing. Discovered in the Reionization Lensing Cluster Survey (RELICS) Hubble Treasury program and companion S-RELICS Spitzer program, this candidate has a lensed H-band magnitude of 25.7+/-0.1 AB mag. With a magnification of \mu~4-7 estimated from our lens models, the de-lensed intrinsic magnitude is 27.6+/-0.3 AB mag, and the half-light radius is r_e<0.8 kpc, both consistent with other z>9 candidates. The inferred stellar mass (log [M* /M_Sun]=9.7^{+0.7}_{-0.5}) and star formation rate (\log [SFR/M_Sun yr^{-1}]=1.3^{+0.2}_{-0.3}) indicate that this candidate is a typical star-forming galaxy on the z>6 SFR-M* relation. We note that three independent lens models predict two counterimages, at least one of which should be of a similar magnitude to the arc, but these counterimages are not yet detected. Counterimages would not be expected if the arc were at lower redshift. However, the only spectral energy distributions capable of fitting the Hubble and Spitzer photometry well at lower redshifts require unphysical combinations of z~2 galaxy properties. The unprecedented lensed size of this z~10 candidate offers the potential for the James Webb Space Telescope to study the geometric and kinematic properties of a galaxy observed 500 Myr after the Big Bang.Comment: 7 pages, 4 figures. Submitted to ApJ Letter

RELICS: The Reionization Lensing Cluster Survey and the Brightest High-z Galaxies

Massive foreground galaxy clusters magnify and distort the light of objects behind them, permitting a view into both the extremely distant and intrinsically faint galaxy populations. We present here the z ~ 6-8 candidate high-redshift galaxies from the Reionization Lensing Cluster Survey (RELICS), a Hubble and Spitzer Space Telescope survey of 41 massive galaxy clusters spanning an area of ≈200 arcmin². These clusters were selected to be excellent lenses, and we find similar high-redshift sample sizes and magnitude distributions as the Cluster Lensing And Supernova survey with Hubble (CLASH). We discover 257, 57, and eight candidate galaxies at z ~ 6, 7, and 8 respectively, (322 in total). The observed (lensed) magnitudes of the z ~ 6 candidates are as bright as AB mag ~23, making them among the brightest known at these redshifts, comparable with discoveries from much wider, blank-field surveys. RELICS demonstrates the efficiency of using strong gravitational lenses to produce high-redshift samples in the epoch of reionization. These brightly observed galaxies are excellent targets for follow-up study with current and future observatories, including the James Webb Space Telescope