115 research outputs found
Expression determinants of mammalian argonaute proteins in mediating gene silencing
RNA interference occurs by two main processes: mRNA site-specific cleavage and non-cleavage-based mRNA degradation or translational repression. Site-specific cleavage is carried out by argonaute-2 (Ago2), while all four mammalian argonaute proteins (Ago1–Ago4) can carry out non-cleavage-mediated inhibition, suggesting that Ago1, Ago3 and Ago4 may have similar but potentially redundant functions. It has been observed that in mammalian tissues, expression of Ago3 and Ago4 is dramatically lower compared with Ago1; however, an optimization of the Ago3 and Ago4 coding sequences to include only the most common codon at each amino acid position was able to augment the expression of Ago3 and Ago4 to levels comparable to that of Ago1 and Ago2. Thus, we examined whether particular sequence features exist in the coding region of Ago3 and Ago4 that may prevent a high level of expression. Swapping specific sub-regions of wild-type and optimized Ago sequence identified the portion of the coding region (nucleotides 1–1163 for Ago-3 and 1–1494 for Ago-4) that is most influential for expression. This finding has implications for the evolutionary conservation of Ago proteins in the mammalian lineage and the biological role that potentially redundant Ago proteins may have
Grouping of UVCB substances with dose-response transcriptomics data from human cell-based assays
The application of in vitro biological assays as new approach methodologies (NAMs) to support grouping of UVCB (unknown or variable composition, complex reaction products, and biological materials) substances has recently been demonstrated. In addition to cell-based phenotyping as NAMs, in vitro transcriptomic profiling is used to gain deeper mechanistic understanding of biological responses to chemicals and to support grouping and read-across. However, the value of gene expression profiling for characterizing complex substances like UVCBs has not been explored. Using 141 petroleum substance extracts, we performed dose-response transcriptomic profiling in human induced pluripotent stem cell (iPSC)-derived hepatocytes, cardiomyocytes, neurons, and endothelial cells, as well as cell lines MCF7 and A375. The goal was to determine whether transcriptomic data can be used to group these UVCBs and to further characterize the molecular basis for in vitro biological responses. We found distinct transcriptional responses for petroleum substances by manufacturing class. Pathway enrichment informed interpretation of effects of substances and UVCB petroleum-class. Transcriptional activity was strongly correlated with concentration of polycyclic aromatic compounds (PAC), especially in iPSC-derived hepatocytes. Supervised analysis using transcriptomics, alone or in combination with bioactivity data collected on these same substances/cells, suggest that transcriptomics data provide useful mechanistic information, but only modest additional value for grouping. Overall, these results further demonstrate the value of NAMs for grouping of UVCBs, identify informative cell lines, and provide data that could be used for justifying selection of substances for further testing that may be required for registration
Integrative Chemical–Biological Grouping of Complex High Production Volume Substances from Lower Olefin Manufacturing Streams
Human cell-based test methods can be used to evaluate potential hazards of mixtures and products of petroleum refining (“unknown or variable composition, complex reaction products, or biological materials” substances, UVCBs). Analyses of bioactivity and detailed chemical characterization of petroleum UVCBs were used separately for grouping these substances; a combination of the approaches has not been undertaken. Therefore, we used a case example of representative high production volume categories of petroleum UVCBs, 25 lower olefin substances from low benzene naphtha and resin oils categories, to determine whether existing manufacturing-based category grouping can be supported. We collected two types of data: nontarget ion mobility spectrometry-mass spectrometry of both neat substances and their organic extracts and in vitro bioactivity of the organic extracts in five human cell types: umbilical vein endothelial cells and induced pluripotent stem cell-derived hepatocytes, endothelial cells, neurons, and cardiomyocytes. We found that while similarity in composition and bioactivity can be observed for some substances, existing categories are largely heterogeneous. Strong relationships between composition and bioactivity were observed, and individual constituents that determine these associations were identified. Overall, this study showed a promising approach that combines chemical composition and bioactivity data to better characterize the variability within manufacturing categories of petroleum UVCBs
Searching for Massive Outflows in Holmberg IX X-1 and NGC 1313 X-1: The Iron K Band
We have analysed all the good quality XMM-Newton data publicly available for
the bright ULXs Holmberg IX X-1 and NGC 1313 X-1, with the aim of searching for
discrete emission or absorption features in the Fe K band that could provide
observational evidence for the massive outflows predicted if these sources are
accreting at substantially super-Eddington rates. We do not find statistically
compelling evidence for any atomic lines, and the limits that are obtained have
interesting consequences. Any features in the immediate Fe K energy band (6-7
keV) must have equivalent widths weaker than ~30 eV for Holmberg IX X-1, and
weaker than ~50 eV for NGC 1313 X-1 (at 99 per cent confidence). In comparison
to the sub-Eddington outflows observed in GRS 1915+105, which imprint iron
absorption features with equivalent widths of ~30 eV, the limits obtained here
appear quite stringent, particularly when Holmberg IX X-1 and NGC 1313 X-1 must
be expelling at least 5-10 times as much material if they host black holes of
similar masses. The difficulty in reconciling these observational limits with
the presence of strong line-of-sight outflows suggests that either these
sources are not launching such outflows, or that they must be directed away
from our viewing angle.Comment: 11 pages, 5 figures, accepted for publication in MNRA
Phase 1 Trial of Antibody NI006 for Depletion of Cardiac Transthyretin Amyloid
BACKGROUND
Transthyretin amyloid (ATTR) cardiomyopathy is a progressive and fatal disease caused by misfolded transthyretin. Despite advances in slowing disease progression, there is no available treatment that depletes ATTR from the heart for the amelioration of cardiac dysfunction. NI006 is a recombinant human anti-ATTR antibody that was developed for the removal of ATTR by phagocytic immune cells.
METHODS
In this phase 1, double-blind trial, we randomly assigned (in a 2:1 ratio) 40 patients with wild-type or variant ATTR cardiomyopathy and chronic heart failure to receive intravenous infusions of either NI006 or placebo every 4 weeks for 4 months. Patients were sequentially enrolled in six cohorts that received ascending doses (ranging from 0.3 to 60 mg per kilogram of body weight). After four infusions, patients were enrolled in an open-label extension phase in which they received eight infusions of NI006 with stepwise increases in the dose. The safety and pharmacokinetic profiles of NI006 were assessed, and cardiac imaging studies were performed.
RESULTS
The use of NI006 was associated with no apparent drug-related serious adverse events. The pharmacokinetic profile of NI006 was consistent with that of an IgG antibody, and no antidrug antibodies were detected. At doses of at least 10 mg per kilogram, cardiac tracer uptake on scintigraphy and extracellular volume on cardiac magnetic resonance imaging, both of which are imaging-based surrogate markers of cardiac amyloid load, appeared to be reduced over a period of 12 months. The median N-terminal pro-B-type natriuretic peptide and troponin T levels also seemed to be reduced.
CONCLUSIONS
In this phase 1 trial of the recombinant human antibody NI006 for the treatment of patients with ATTR cardiomyopathy and heart failure, the use of NI006 was associated with no apparent drug-related serious adverse events. (Funded by Neurimmune; NI006-101 ClinicalTrials.gov number, NCT04360434.)
A human antibody against pathologic IAPP aggregates protects beta cells in type 2 diabetes models
In patients with type 2 diabetes, pancreatic beta cells progressively degenerate and gradually lose their ability to produce insulin and regulate blood glucose. Beta cell dysfunction and loss is associated with an accumulation of aggregated forms of islet amyloid polypeptide (IAPP) consisting of soluble prefibrillar IAPP oligomers as well as insoluble IAPP fibrils in pancreatic islets. Here, we describe a human monoclonal antibody selectively targeting IAPP oligomers and neutralizing IAPP aggregate toxicity by preventing membrane disruption and apoptosis in vitro. Antibody treatment in male rats and mice transgenic for human IAPP, and human islet-engrafted mouse models of type 2 diabetes triggers clearance of IAPP oligomers resulting in beta cell protection and improved glucose control. These results provide new evidence for the pathological role of IAPP oligomers and suggest that antibody-mediated removal of IAPP oligomers could be a pharmaceutical strategy to support beta cell function in type 2 diabetes
Rational F-Theory GUTs without exotics
We construct F-theory GUT models without exotic matter, leading to the MSSM
matter spectrum with potential singlet extensions. The interplay of engineering
explicit geometric setups, absence of four-dimensional anomalies, and realistic
phenomenology of the couplings places severe constraints on the allowed local
models in a given geometry. In constructions based on the spectral cover we
find no model satisfying all these requirements. We then provide a survey of
models with additional U(1) symmetries arising from rational sections of the
elliptic fibration in toric constructions and obtain phenomenologically
appealing models based on SU(5) tops. Furthermore we perform a bottom-up
exploration beyond the toric section constructions discussed in the literature
so far and identify benchmark models passing all our criteria, which can serve
as a guideline for future geometric engineering.Comment: 27 Pages, 1 Figur
Black hole spin: theory and observation
In the standard paradigm, astrophysical black holes can be described solely
by their mass and angular momentum - commonly referred to as `spin' - resulting
from the process of their birth and subsequent growth via accretion. Whilst the
mass has a standard Newtonian interpretation, the spin does not, with the
effect of non-zero spin leaving an indelible imprint on the space-time closest
to the black hole. As a consequence of relativistic frame-dragging, particle
orbits are affected both in terms of stability and precession, which impacts on
the emission characteristics of accreting black holes both stellar mass in
black hole binaries (BHBs) and supermassive in active galactic nuclei (AGN).
Over the last 30 years, techniques have been developed that take into account
these changes to estimate the spin which can then be used to understand the
birth and growth of black holes and potentially the powering of powerful jets.
In this chapter we provide a broad overview of both the theoretical effects of
spin, the means by which it can be estimated and the results of ongoing
campaigns.Comment: 55 pages, 5 figures. Published in: "Astrophysics of Black Holes -
From fundamental aspects to latest developments", Ed. Cosimo Bambi, Springer:
Astrophysics and Space Science Library. Additional corrections mad
Combinations of single-top-quark production cross-section measurements and vertical bar f(LV)V(tb)vertical bar determinations at root s=7 and 8 TeV with the ATLAS and CMS experiments
This paper presents the combinations of single-top-quark production cross-section measurements by the ATLAS and CMS Collaborations, using data from LHC proton-proton collisions at = 7 and 8 TeV corresponding to integrated luminosities of 1.17 to 5.1 fb(-1) at = 7 TeV and 12.2 to 20.3 fb(-1) at = 8 TeV. These combinations are performed per centre-of-mass energy and for each production mode: t-channel, tW, and s-channel. The combined t-channel cross-sections are 67.5 +/- 5.7 pb and 87.7 +/- 5.8 pb at = 7 and 8 TeV respectively. The combined tW cross-sections are 16.3 +/- 4.1 pb and 23.1 +/- 3.6 pb at = 7 and 8 TeV respectively. For the s-channel cross-section, the combination yields 4.9 +/- 1.4 pb at = 8 TeV. The square of the magnitude of the CKM matrix element V-tb multiplied by a form factor f(LV) is determined for each production mode and centre-of-mass energy, using the ratio of the measured cross-section to its theoretical prediction. It is assumed that the top-quark-related CKM matrix elements obey the relation |V-td|, |V-ts| << |V-tb|. All the |f(LV)V(tb)|(2) determinations, extracted from individual ratios at = 7 and 8 TeV, are combined, resulting in |f(LV)V(tb)| = 1.02 +/- 0.04 (meas.) +/- 0.02 (theo.). All combined measurements are consistent with their corresponding Standard Model predictions.Peer reviewe
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