Neuroblastoma is a paediatric cancer which originates from precursor cells of the sympathetic nervous system and accounts for 15% of childhood cancer mortalities. With regards to the role of miRNAs ...in neuroblastoma, miR-34a, mapping to a chromosome 1p36 region that is commonly deleted, has been found to act as a tumor suppressor through targeting of numerous genes associated with cell proliferation and apoptosis.
A synthetic miR-34a (or negative control) precursor molecule was transfected into NB1691luc and SK-N-ASluc neuroblastoma cells. Quantitative PCR was used to verify increased miR-34a levels in NB1691luc and SK-N-ASluc cell lines prior to in vitro and in vivo analysis. In vitro analysis of the effects of miR-34a over expression on cell growth, cell cycle and phosphoprotein activation in signal transduction pathways was performed. Neuroblastoma cells over expressing miR-34a were injected retroperitoneally into immunocompromised CB17-SCID mice and tumor burden was assessed over a 21 day period by measuring bioluminescence (photons/sec/cm²).
Over expression of miR-34a in both NB1691luc and SK-N-ASluc neuroblastoma cell lines led to a significant decrease in cell number relative to premiR-negative control treated cells over a 72 hour period. Flow cytometry results indicated that miR-34a induced cell cycle arrest and subsequent apoptosis activation. Phosphoprotein analysis highlighted key elements involved in signal transduction, whose activation was dysregulated as a result of miR-34a introduction into cells. As a potential mechanism of miR-34a action on phosphoprotein levels, we demonstrate that miR-34a over-expression results in a significant reduction of MAP3K9 mRNA and protein levels. Although MAP3K9 is a predicted target of miR-34a, direct targeting could not be validated with luciferase reporter assays. Despite this fact, any functional effects of reduced MAP3K9 expression as a result of miR-34a would be expected to be similar regardless of the mechanism involved. Most notably, in vivo studies showed that tumor growth was significantly repressed after exogenous miR-34a administration in retroperitoneal neuroblastoma tumors.
We demonstrate for the first time that miR-34a significantly reduces tumor growth in an in vivo orthotopic murine model of neuroblastoma and identified novel effects that miR-34a has on phospho-activation of key proteins involved with apoptosis.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
The Taiwanese-American Occultation Survey (TAOS) aims to detect serendipitous occultations of stars by small (~1 km diameter) objects in the Kuiper Belt and beyond. Such events are very rare (<10 ...super(-3) events per star per year) and short in duration (~200 ms), so many stars must be monitored at a high readout cadence. TAOS monitors typically ~500 stars simultaneously at a 5 Hz readout cadence with four telescopes located at Lulin Observatory in central Taiwan. In this paper, we report the results of the search for small Kuiper Belt objects (KBOs) in seven years of data. No occultation events were found, resulting in a 95% c.l. upper limit on the slope of the faint end of the KBO size distribution of q - 3.34-3.82, depending on the surface density at the break in the size distribution at a diameter of about 90 km.
Achieving maximum scientific results from the overwhelming volume of astronomical data to be acquired over the next few decades demands novel, fully automatic methods of data analysis. Here we ...concentrate on eclipsing binary (EB) stars, a prime source of astrophysical information, of which only some hundreds have been rigorously analyzed, but whose numbers will reach millions in a decade. We describe the artificial neural network (ANN) approach which is able to surmount the human bottleneck and permit EB-based scientific yield to keep pace with future data rates. The ANN, following training on a sample of 33,235 model light curves, outputs a set of approximate model parameters image, and image for each input light curve data set. The obtained parameters can then be readily passed to sophisticated modeling engines. We also describe a novel method polyfit for preprocessing observational light curves before inputting their data to the ANN and present the results and analysis of testing the approach on synthetic data and on real data including 50 binaries from the Catalog and Atlas of Eclipsing Binaries (CALEB) database and 2580 light curves from OGLE survey data. The success rate, defined by less than a 10% error in the network output parameter values, is approximately 90% for the OGLE sample and close to 100% for the CALEB sample-sufficient for a reliable statistical analysis. The code is made available to the public. Our approach is applicable to EB light curves of all classes; this first paper in the eclipsing binaries via artificial intelligence (EBAI) series focuses on detached EBs, which is the class most challenging for this approach.
The acquisition of multidrug resistance is a major impediment to the successful treatment of neuroblastoma, a clinically heterogeneous cancer accounting for ∼15% of all pediatric cancer deaths. The ...MYCN transcription factor, whose gene is amplified in ∼30% of high‐risk neuroblastoma cases, influences drug resistance by regulating a cadre of genes, including those involved with drug efflux, however, other high‐risk subtypes of neuroblastoma lacking MYCN amplification, such as those with chromosome 11q deletions, also acquire multidrug resistance. To elucidate additional mechanisms involved with drug resistance in non‐MYCN amplified tumour cells, an SK‐N‐AS subline (SK‐N‐AsCis24) that is significantly resistant to cisplatin and cross resistant to etoposide was developed through a pulse‐selection process. High resolution aCGH analysis of SK‐N‐AsCis24 revealed a focal gain on chromosome 5 containing the coding sequence for the neural apoptosis inhibitory protein (NAIP). Significant overexpression of NAIP mRNA and protein was documented, while experimental modulation of NAIP levels in both SK‐N‐AsCis24 and in parental SK‐N‐AS cells confirmed that NAIP was responsible for the drug resistant phenotype by apoptosis inhibition. Furthermore, a decrease in the NAIP targeting microRNA, miR‐520f, was also demonstrated to be partially responsible for increased NAIP levels in SK‐N‐AsCis24. Interestingly, miR‐520f levels were determined to be significantly lower in postchemotherapy treatment tumours relative to matched prechemotherapy samples, consistent with a role for this miRNA in the acquisition of drug resistance in vivo, potentially through decreased NAIP targeting. Our findings provide biological novel insight into neuroblastoma drug‐resistance and have implications for future therapeutic research.
What's new?
Just under one‐third of high‐risk neuroblastomas, in which drug resistance is a central feature, involve amplification of the MYCN gene. Resistance in remaining high‐risk tumors may be determined by any of several non‐MYCN amplification mechanisms. Here, analysis of an SK‐N‐AS subline (SK‐N‐AsCis24) lacking MYCN amplification but resistant to cisplatin and etoposide reveals a link between the development of drug resistance via apoptotic inhibition and the neural apoptosis inhibitory protein (NAIP) and its regulatory microRNA, miR‐520f. NAIP upregulation was associated with DNA copy number gain on chromosome 5 and down‐regulation of miRNA‐520f. MiR‐520f was discovered to be down‐regulated post‐chemotherapy.
Abstract
We present a new pipeline based on the Support Vector Machine algorithm to confirm the detection and perform classification of small solar system objects by serendipitous stellar ...occultations. This pipeline is designed to analyze light curves and to identify the occultation events and the classification of the occulting bodies according to their size, typically from a fraction to a few kilometers, and their distance from the Sun, typically a few tens of astronomical units. The input light curves for this pipeline were obtained from the event simulator for the Trans-Neptunian Automated Occultation Survey (TAOS II). We explore parameters affecting occultation light curves such as spectral type, apparent magnitude and finite angular size of the occulted star, angle from opposition, and readout cadence for the observations; also we assumed a Poisson noise distribution as expected from the TAOS II project. We find that occultation events, especially by trans-Neptunian objects with diameters ≥2 km are detected with 99.99%, 99.53%, and 86% efficiency for stars with a visual apparent magnitude of 12, 14, and 16, respectively at 0.05 s of exposure time. In terms of size and distance classification the overall accuracy is 94%. However, for smaller objects the confirmation and classification depends mostly upon the signal-to-noise ratio.
We present a new occultation event simulator for the Trans-Neptunian Automated Occultation Survey (TAOS II). We have developed a method to compute occultation shadows by small objects with ...non-circular apparent shapes (as may result from an intrinsic morphology or from the projection of a contact binary). The new simulator calculates diffraction features in the occultation shadows, as well as resulting light curves as would be measured by the TAOS II survey system. We include effects such as the spectral type and finite angular size of the occulted star. We find that occultation events, especially by Trans-Neptunian Objects with diameters ∼3 km may be misidentified or mischaracterized when not taking non-spherical shapes into account.
Using 7 yr of MACHO survey data, we present a new determination of the optical depth to microlensing toward the Galactic bulge. We select the sample of 62 microlensing events (60 unique) on clump ...giant sources and perform a detailed efficiency analysis. We use only the clump giant sources because these are bright bulge stars and are not as strongly affected by blending as other events. Using a subsample of 42 clump events concentrated in an area of 4.5 deg super(2) with 739,000 clump giant stars, we find t = 2.17 super(+) sub(-) super(0) sub(0) super(.) sub(.) super(4) sub(3) super(7) sub(8) x 10 super(-6) at (l,b) = (1.50, - 2.68), somewhat smaller than found in most previous MACHO studies but in excellent agreement with recent theoretical predictions. We also present the optical depth in each of the 19 fields in which we detected events and find limits on optical depth for fields with no events. The errors in optical depth in individual fields are dominated by Poisson noise. We measure optical depth gradients of (1.06 c 0.71) x 10 super(-6) deg super(-1) and (0.29 c 0.43) x 10 super(-6) deg super(-1) in the Galactic latitude b and longitude l directions, respectively. Finally, we discuss the possibility of anomalous duration distribution of events in the field 104 centered on (l,b) = (3.11, - 3.01), as well as investigate spatial clustering of events in all fields.
The serendipitous detection of stellar occultations by outer solar system objects is a powerful method for ascertaining the small end (r 15 km) of the size distribution of Kuiper Belt objects and may ...potentially allow the exploration of objects as far out as the Oort Cloud. The design and implementation of an occultation survey is aided by a detailed understanding of how diffraction and observational parameters affect the detection of occultation events. In this study, stellar occultations are simulated, accounting for diffraction effects, finite source sizes, finite bandwidths, stellar spectra, sampling, and signal-to-noise ratios. Finally, the possibility of detecting small outer solar system objects from the Kuiper Belt all the way out to the Oort Cloud is explored for three photometric systems: a proposed space telescope, Whipple, the Taiwanese-American Occultation Survey, and the MMT.
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