644 research outputs found
Structural and Functional analysis of the GABARAP interaction motif (GIM)
© 2017 The Authors. Published under the terms of the CC BY 4.0 license. Through the canonical LC3 interaction motif (LIR), [W/F/Y]-X 1 -X 2 -[I/L/V], protein complexes are recruited to autophagosomes to perform their functions as either autophagy adaptors or receptors. How these adaptors/receptors selectively interact with either LC3 or GABARAP families remains unclear. Herein, we determine the range of selectivity of 30 known core LIR motifs towards individual LC3s and GABARAPs. From these, we define a GABARAP Interaction Motif (GIM) sequence ([W/F] -[V/I]-X 2 -V) that the adaptor protein PLEKHM1 tightly conforms to. Using biophysical and structural approaches, we show that the PLEKHM1-LIR is indeed 11-fold more specific for GABARAP than LC3B. Selective mutation of the X 1 and X 2 positions either completely abolished the interaction with all LC3 and GABARAPs or increased PLEKHM1-GIM selectivity 20-fold towards LC3B. Finally, we show that conversion of p62/SQSTM1, FUNDC1 and FIP200 LIRs into our newly defined GIM, by introducing two valine residues, enhances their interaction with endogenous GABARAP over LC3B. The identification of a GABARAP-specific interaction motif will aid the identification and characterization of the expanding array of autophagy receptor and adaptor proteins and their in vivo functions
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Antiproliferative effects of masitinib and imatinib against canine oral fibrosarcoma in vitro
Background:
Canine oral fibrosarcoma (COF) is one of the most common oral tumors in dogs and carries a guarded prognosis due to a lack of effective systemic therapeutic options. Mastinib and imatinib are two commonly used tyrosine kinase inhibitors (TKIs) in veterinary oncology but their potential efficacy against COF is uncharacterized. To begin investigating the rationale for use of these TKIs against COF, the present study tested for the presence TKI targets PDGFR-α, PDGFR-β, Kit, and VEGFR-2 and examined the in vitro effects on cell viability after TKI treatment alone or with doxorubicin.
Immunohistochemistry for PDGFR-α, PDGFR-β, Kit, and VEGFR-2 was performed in 6 COF tumor biopsies. Presence of these same receptors within 2 COF cell lines was probed by reverse transcription-polymerase chain reaction and, for those with mRNA detected, confirmed via western blot. Effects on cell viability were assessed using an MTS assay after masitinib or imatinib treatment alone (0-100 μM), or in combination with doxorubicin (0-3000 nM doxorubicin). Anti-PDGFRB siRNA knockdown was performed and the effect on cell viability quantified.
Results:
Expression of the TKI targets evaluated was similar between the 2 COF cell lines and the 6 COF tumor biopsies: PDGFR-α and PDGFR-β were detected in neoplastic cells from most COF tumor biopsies (5/6 and 6/6, respectively) and were present in both COF cell lines; KIT and KDR were not detected in any sample. Masitinib and imatinib IC50 values ranged from 7.9–33.4 μM, depending on the specific TKI and cell line tested. The addition of doxorubicin resulted in synergistic cytotoxicity with both TKIs. Anti-PDGFRB siRNA transfection reduced PDGFR-β protein expression by 77 % and 67 % and reduced cell viability by 24 % (p < 0.0001) and 28 % (0 = 0.0003) in the two cell lines, respectively.
Conclusions:
These results provide rationale for further investigation into the use of TKIs, possibly in combination with doxorubicin, as treatment options for COF.Keywords: Masitinib, Oral fibrosarcoma, Imatinib, Platelet-derived growth factor receptor, Do
Life-threatening intoxication with methylene bis(thiocyanate): clinical picture and pitfalls. A case report
BACKGROUND: Methylene bis(thiocyanate) (MBT) is a microbiocidal agent mainly used in industrial water cooling systems and paper mills as an inhibitor of algae, fungi, and bacteria. CASE PRESENTATION: We describe the first case of severe intoxication following inhalation of powder in an industrial worker. Profound cyanosis and respiratory failure caused by severe methemoglobinemia developed within several minutes. Despite immediate admission to the intensive care unit, where mechanical ventilation and hemodialysis for toxin elimination were initiated, multi-organ failure involving liver, kidneys, and lungs developed. While liver failure was leading, the patient was successfully treated with the MARS (molecular adsorbent recirculating system) procedure. CONCLUSION: Intoxication with MBT is a potentially life-threatening intoxication causing severe methemoglobinemia and multi-organ failure. Extracorporeal liver albumin dialysis (MARS) appears to be an effective treatment to allow recovery of hepatic function
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Mechanism-based classification of PAH mixtures to predict carcinogenic potential
We have previously shown that relative potency factors and DNA adduct measurements are inadequate for predicting carcinogenicity of certain polycyclic aromatic hydrocarbons (PAHs) and PAH mixtures, particularly those that function through alternate pathways or exhibit greater
promotional activity compared to benzo[a]pyrene (BaP). Therefore, we developed a pathway
based approach for classification of tumor outcome after dermal exposure to PAH/mixtures.
FVB/N mice were exposed to dibenzo[def,p]chrysene (DBC), BaP or environmental PAH
mixtures (Mix 1-3) following a two-stage initiation/promotion skin tumor protocol. Resulting
tumor incidence could be categorized by carcinogenic potency as
DBC>>BaP=Mix2=Mix3>Mix1=Control, based on statistical significance. Gene expression
profiles measured in skin of mice collected 12 h post-initiation were compared to tumor outcome
for identification of short-term bioactivity profiles. A Bayesian integration model was utilized to
identify biological pathways predictive of PAH carcinogenic potential during initiation.
Integration of probability matrices from four enriched pathways (p<0.05) for DNA damage,
apoptosis, response to chemical stimulus and interferon gamma signaling resulted in the highest
classification accuracy with leave-one-out cross validation. This pathway-driven approach was
successfully utilized to distinguish early regulatory events during initiation prognostic for tumor
outcome and provides proof-of-concept for using short-term initiation studies to classify
carcinogenic potential of environmental PAH mixtures. These data further provide a ‘source-to
outcome’ model that could be used to predict PAH interactions during tumorigenesis and provide
an example of how mode-of-action based risk assessment could be employed for environmental
PAH mixtures.This is an author's peer-reviewed final manuscript, as accepted by the publisher. The published article is copyrighted by Oxford University Press and can be found at: http://toxsci.oxfordjournals.org/.Keywords: mixtures, toxicogenomics, skin cancer, polycyclic aromatic hydrocarbons, modelin
High-E_T dijet photoproduction at HERA
The cross section for high-E_T dijet production in photoproduction has been
measured with the ZEUS detector at HERA using an integrated luminosity of 81.8
pb-1. The events were required to have a virtuality of the incoming photon,
Q^2, of less than 1 GeV^2 and a photon-proton centre-of-mass energy in the
range 142 < W < 293 GeV. Events were selected if at least two jets satisfied
the transverse-energy requirements of E_T(jet1) > 20 GeV and E_T(jet2) > 15 GeV
and pseudorapidity requirements of -1 < eta(jet1,2) < 3, with at least one of
the jets satisfying -1 < eta(jet) < 2.5. The measurements show sensitivity to
the parton distributions in the photon and proton and effects beyond
next-to-leading order in QCD. Hence these data can be used to constrain further
the parton densities in the proton and photon.Comment: 36 pages, 13 figures, 20 tables, including minor revisions from
referees. Accepted by Phys. Rev.
An NLO QCD analysis of inclusive cross-section and jet-production data from the ZEUS experiment
The ZEUS inclusive differential cross-section data from HERA, for charged and
neutral current processes taken with e+ and e- beams, together with
differential cross-section data on inclusive jet production in e+ p scattering
and dijet production in \gamma p scattering, have been used in a new NLO QCD
analysis to extract the parton distribution functions of the proton. The input
of jet data constrains the gluon and allows an accurate extraction of
\alpha_s(M_Z) at NLO;
\alpha_s(M_Z) = 0.1183 \pm 0.0028(exp.) \pm 0.0008(model)
An additional uncertainty from the choice of scales is estimated as \pm
0.005. This is the first extraction of \alpha_s(M_Z) from HERA data alone.Comment: 37 pages, 14 figures, to be submitted to EPJC. PDFs available at
http://durpdg.dur.ac.uk/hepdata in LHAPDFv
Measurement of (anti)deuteron and (anti)proton production in DIS at HERA
The first observation of (anti)deuterons in deep inelastic scattering at HERA
has been made with the ZEUS detector at a centre-of-mass energy of 300--318 GeV
using an integrated luminosity of 120 pb-1. The measurement was performed in
the central rapidity region for transverse momentum per unit of mass in the
range 0.3<p_T/M<0.7. The particle rates have been extracted and interpreted in
terms of the coalescence model. The (anti)deuteron production yield is smaller
than the (anti)proton yield by approximately three orders of magnitude,
consistent with the world measurements.Comment: 26 pages, 9 figures, 5 tables, submitted to Nucl. Phys.
Limits on the effective quark radius from inclusive scattering at HERA
The high-precision HERA data allows searches up to TeV scales for Beyond the
Standard Model contributions to electron-quark scattering. Combined
measurements of the inclusive deep inelastic cross sections in neutral and
charged current scattering corresponding to a luminosity of around 1
fb have been used in this analysis. A new approach to the beyond the
Standard Model analysis of the inclusive data is presented; simultaneous
fits of parton distribution functions together with contributions of "new
physics" processes were performed. Results are presented considering a finite
radius of quarks within the quark form-factor model. The resulting 95% C.L.
upper limit on the effective quark radius is cm.Comment: 10 pages, 4 figures, accepted by Phys. Lett.
Measurement of the cross-section ratio sigma_{psi(2S)}/sigma_{J/psi(1S)} in deep inelastic exclusive ep scattering at HERA
The exclusive deep inelastic electroproduction of and
at an centre-of-mass energy of 317 GeV has been studied with the ZEUS
detector at HERA in the kinematic range GeV,
GeV and GeV, where is the photon virtuality, is the
photon-proton centre-of-mass energy and is the squared four-momentum
transfer at the proton vertex. The data for GeV were taken in
the HERA I running period and correspond to an integrated luminosity of 114
pb. The data for GeV are from both HERA I and HERA II
periods and correspond to an integrated luminosity of 468 pb. The decay
modes analysed were and for the
and for the . The cross-section ratio
has been measured as a function of
and . The results are compared to predictions of QCD-inspired
models of exclusive vector-meson production.Comment: 24 pages, 8 figure
Measurement of event shapes in deep inelastic scattering at HERA
Inclusive event-shape variables have been measured in the current region of
the Breit frame for neutral current deep inelastic ep scattering using an
integrated luminosity of 45.0 pb^-1 collected with the ZEUS detector at HERA.
The variables studied included thrust, jet broadening and invariant jet mass.
The kinematic range covered was 10 < Q^2 < 20,480 GeV^2 and 6.10^-4 < x < 0.6,
where Q^2 is the virtuality of the exchanged boson and x is the Bjorken
variable. The Q dependence of the shape variables has been used in conjunction
with NLO perturbative calculations and the Dokshitzer-Webber non-perturbative
corrections (`power corrections') to investigate the validity of this approach.Comment: 7+25 pages, 6 figure
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