1,264 research outputs found
A Human Islet Cell-Culture System for High-Throuput screening.
A small-molecule inducer of beta-cell proliferation in human islets represents a potential regeneration strategy for treating type 1 diabetes. However, the lack of suitable human beta cell lines makes such a discovery a challenge. Here, we adapted an islet cell culture system to high-throughput screening to identify such small molecules. We prepared microtiter plates containing extracellular matrix from a human bladder carcinoma cell line. Dissociated human islets were seeded onto these plates, cultured for up to 7 days, and assessed for proliferation by simultaneous Ki67 and C-peptide immunofluorescence. Importantly, this environment preserved beta-cell physiological function, as measured by glucose-stimulated insulin secretion. Adenoviral overexpression of cdk-6 and cyclin D(1), known inducers of human beta cell proliferation, was used as a positive control in our assay. This induction was inhibited by cotreatment with rapamycin, an immunosuppressant often used in islet transplantation. We then performed a pilot screen of 1280 compounds, observing some phenotypic effects on cells. This high-throughput human islet cell culture method can be used to assess various aspects of beta-cell biology on a relatively large number of compounds
High-throughput identification of genotype-specific cancer vulnerabilities in mixtures of barcoded tumor cell lines.
Hundreds of genetically characterized cell lines are available for the discovery of genotype-specific cancer vulnerabilities. However, screening large numbers of compounds against large numbers of cell lines is currently impractical, and such experiments are often difficult to control. Here we report a method called PRISM that allows pooled screening of mixtures of cancer cell lines by labeling each cell line with 24-nucleotide barcodes. PRISM revealed the expected patterns of cell killing seen in conventional (unpooled) assays. In a screen of 102 cell lines across 8,400 compounds, PRISM led to the identification of BRD-7880 as a potent and highly specific inhibitor of aurora kinases B and C. Cell line pools also efficiently formed tumors as xenografts, and PRISM recapitulated the expected pattern of erlotinib sensitivity in vivo
A transformed view of cyclosporine
Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/62591/1/397471a0.pd
Many-body localization in a quantum simulator with programmable random disorder
When a system thermalizes it loses all local memory of its initial
conditions. This is a general feature of open systems and is well described by
equilibrium statistical mechanics. Even within a closed (or reversible) quantum
system, where unitary time evolution retains all information about its initial
state, subsystems can still thermalize using the rest of the system as an
effective heat bath. Exceptions to quantum thermalization have been predicted
and observed, but typically require inherent symmetries or noninteracting
particles in the presence of static disorder. The prediction of many-body
localization (MBL), in which disordered quantum systems can fail to thermalize
in spite of strong interactions and high excitation energy, was therefore
surprising and has attracted considerable theoretical attention. Here we
experimentally generate MBL states by applying an Ising Hamiltonian with
long-range interactions and programmably random disorder to ten spins
initialized far from equilibrium. We observe the essential signatures of MBL:
memory retention of the initial state, a Poissonian distribution of energy
level spacings, and entanglement growth in the system at long times. Our
platform can be scaled to higher numbers of spins, where detailed modeling of
MBL becomes impossible due to the complexity of representing such entangled
quantum states. Moreover, the high degree of control in our experiment may
guide the use of MBL states as potential quantum memories in naturally
disordered quantum systems.Comment: 9 pages, 9 figure
A massive, quiescent galaxy at redshift of z=3.717
In the early Universe finding massive galaxies that have stopped forming
stars present an observational challenge as their rest-frame ultraviolet
emission is negligible and they can only be reliably identified by extremely
deep near-infrared surveys. These have revealed the presence of massive,
quiescent early-type galaxies appearing in the universe as early as z2,
an epoch 3 Gyr after the Big Bang. Their age and formation processes have now
been explained by an improved generation of galaxy formation models where they
form rapidly at z3-4, consistent with the typical masses and ages derived
from their observations. Deeper surveys have now reported evidence for
populations of massive, quiescent galaxies at even higher redshifts and earlier
times, however the evidence for their existence, and redshift, has relied
entirely on coarsely sampled photometry. These early massive, quiescent
galaxies are not predicted by the latest generation of theoretical models.
Here, we report the spectroscopic confirmation of one of these galaxies at
redshift z=3.717 with a stellar mass of 1.710 M whose
absorption line spectrum shows no current star-formation and which has a
derived age of nearly half the age of the Universe at this redshift. The
observations demonstrates that the galaxy must have quickly formed the majority
of its stars within the first billion years of cosmic history in an extreme and
short starburst. This ancestral event is similar to those starting to be found
by sub-mm wavelength surveys pointing to a possible connection between these
two populations. Early formation of such massive systems is likely to require
significant revisions to our picture of early galaxy assembly.Comment: 6 pages, 7 figures. This is the final preprint corresponding closely
to the published version. Uploaded 6 months after publication in accordance
with Nature polic
Mosaic DNA imports with interspersions of recipient sequence after natural transformation of Helicobacter pylori
Helicobacter pylori colonizes the gastric mucosa of half of the human population, causing gastritis, ulcers, and cancer. H. pylori
is naturally competent for transformation by exogenous DNA, and recombination during mixed infections of one stomach
with multiple H. pylori strains generates extensive allelic diversity. We developed an in vitro transformation protocol to study
genomic imports after natural transformation of H. pylori. The mean length of imported fragments was dependent on the
combination of donor and recipient strain and varied between 1294 bp and 3853 bp. In about 10% of recombinant clones, the
imported fragments of donor DNA were interrupted by short interspersed sequences of the recipient (ISR) with a mean length
of 82 bp. 18 candidate genes were inactivated in order to identify genes involved in the control of import length and
generation of ISR. Inactivation of the antimutator glycosylase MutY increased the length of imports, but did not have a
significant effect on ISR frequency. Overexpression of mutY strongly increased the frequency of ISR, indicating that MutY, while
not indispensable for ISR formation, is part of at least one ISR-generating pathway. The formation of ISR in H. pylori increases
allelic diversity, and contributes to the uniquely low linkage disequilibrium characteristic of this pathogen
Mapping Dynamic Histone Acetylation Patterns to Gene Expression in Nanog-depleted Murine Embryonic Stem Cells
Embryonic stem cells (ESC) have the potential to self-renew indefinitely and
to differentiate into any of the three germ layers. The molecular mechanisms
for self-renewal, maintenance of pluripotency and lineage specification are
poorly understood, but recent results point to a key role for epigenetic
mechanisms. In this study, we focus on quantifying the impact of histone 3
acetylation (H3K9,14ac) on gene expression in murine embryonic stem cells. We
analyze genome-wide histone acetylation patterns and gene expression profiles
measured over the first five days of cell differentiation triggered by
silencing Nanog, a key transcription factor in ESC regulation. We explore the
temporal and spatial dynamics of histone acetylation data and its correlation
with gene expression using supervised and unsupervised statistical models. On a
genome-wide scale, changes in acetylation are significantly correlated to
changes in mRNA expression and, surprisingly, this coherence increases over
time. We quantify the predictive power of histone acetylation for gene
expression changes in a balanced cross-validation procedure. In an in-depth
study we focus on genes central to the regulatory network of Mouse ESC,
including those identified in a recent genome-wide RNAi screen and in the
PluriNet, a computationally derived stem cell signature. We find that compared
to the rest of the genome, ESC-specific genes show significantly more
acetylation signal and a much stronger decrease in acetylation over time, which
is often not reflected in an concordant expression change. These results shed
light on the complexity of the relationship between histone acetylation and
gene expression and are a step forward to dissect the multilayer regulatory
mechanisms that determine stem cell fate.Comment: accepted at PLoS Computational Biolog
Pathophysiology of ANCA-Associated Small Vessel Vasculitis
Antineutrophil cytoplasmic autoantibodies (ANCAs) directed to proteinase 3 (PR3-ANCA) or myeloperoxidase (MPO-ANCA) are strongly associated with the ANCA-associated vasculitides—Wegener’s granulomatosis, microscopic polyangiitis, and Churg-Strauss syndrome. Clinical observations, including the efficacy of B-cell depletion via rituximab treatment, support—but do not prove—a pathogenic role for ANCA in the ANCA-associated vasculitides. In vitro experimental studies show that the interplay of ANCA, neutrophils, the alternative pathway of the complement system, and endothelial cells could result in lysis of the endothelium. A pathogenic role for MPO-ANCA is strongly supported by in vivo experimental studies in mice and rats, which also elucidate the pathogenic mechanisms involved in lesion development. Unfortunately, an animal model for PR3-ANCA–associated Wegener’s granulomatosis is not yet available. Here, cellular immunity appears to play a major role as well, particularly via interleukin-17–producing T cells, in line with granulomatous inflammation in the lesions. Finally, microbial factors, in particular Staphylococcus aureus and gram-negative bacteria, seem to be involved in disease induction and expression, but further studies are needed to define their precise role in disease development
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