Despite intense interest in discovering drugs that cause G-protein-coupled receptors (GPCRs) to selectively stimulate or block arrestin signalling, the structural mechanism of receptor-mediated ...arrestin activation remains unclear
. Here we reveal this mechanism through extensive atomic-level simulations of arrestin. We find that the receptor's transmembrane core and cytoplasmic tail-which bind distinct surfaces on arrestin-can each independently stimulate arrestin activation. We confirm this unanticipated role of the receptor core, and the allosteric coupling between these distant surfaces of arrestin, using site-directed fluorescence spectroscopy. The effect of the receptor core on arrestin conformation is mediated primarily by interactions of the intracellular loops of the receptor with the arrestin body, rather than the marked finger-loop rearrangement that is observed upon receptor binding. In the absence of a receptor, arrestin frequently adopts active conformations when its own C-terminal tail is disengaged, which may explain why certain arrestins remain active long after receptor dissociation. Our results, which suggest that diverse receptor binding modes can activate arrestin, provide a structural foundation for the design of functionally selective ('biased') GPCR-targeted ligands with desired effects on arrestin signalling.
En este informe se presentan datos sobre el comportamiento de herbicidas selectivos en el cultivo de trigo. La mezcla de Picloram + 2,4-D (28,08 + 300 g/ha) mostró tendencia a disminuir el ...rendimiento en la medida en que se atrasó la aplicación, aunque esa disminución sólo fue significativa cuando se aplic6 en el estado aristas visibles. Igual tendencia, pero con menores daños fue puesta de manifiesto por la mezcla Dicamba + 2,4-D (57,71 + 300 g/ha). El Bromoxynil (454,75 g/ha) fue el herbicida que provocó menores daños. El componente granos/espiga explicó en mayor medida las variaciones de rendimiento provocadas por la utilización de herbicidas. La comunidad de malezas presentes no produjo daños. Dirección: F. GARCIA y O. RUBIOLO. Prof. Adj. y Asoc. De Terapéutica vegetal y Cerealicultura, respectivamente.
TP53 mutations are frequent in non-small cell lung cancer (NSCLC) and have been associated with poor outcome. The prognostic and predictive relevance of EGFR/TP53 co-mutations in NSCLC is ...controversial. We analyzed lung tissue specimens from 70 patients with NSCLC using next-generation sequencing to determine EGFR and TP53 status and the association between these status with baseline patient and tumor characteristics, adjuvant treatments, relapse, and progression-free (PFS) and overall survival (OS) after surgical resection. We found the EGFR mutation in 32.9% of patients (20% classical mutations and 12.9% uncommon mutations). TP53 missense mutations occurred in 25.7% and TP53/EGFR co-mutations occurred in 43.5% of patients. Stage after surgical resection was significantly associated with OS (P=0.028). We identified an association between progression-free survival and poor outcome in patients with distant metastases (P=0.007). We found a marginally significant difference in OS between genders (P=0.057) and between mutant and wild type TP53 (P=0.079). In univariate analysis, distant metastases (P=0.027), pathological stage (IIIA-IIIB vs I-II; P=0.028), and TP53 status (borderline significance between wild type and mutant; P=0.079) influenced OS. In multivariable analysis, a significant model for high risk of death and poor OS (P=0.029) selected patients in stage IIIA-IIIB, with relapse and distant metastases, non-responsive to platin-based chemotherapy and erlotinib, with tumors harboring EGFR uncommon mutations, with TP53 mutant, and with EGFR/TP53 co-mutations. Our study suggested that TP53 mutation tends to confer poor survival and a potentially negative predictive effect associated with a non-response to platinum-based chemotherapy and erlotinib in early-stage resected EGFR-mutated NSCLC.
Intratumoral similarities and differences between large-cell neuroendocrine carcinomas (LCNECs) and small-cell lung carcinomas (SCLCs) are determined partially by the Notch signaling pathway, which ...controls the switch from neuroendocrine to slight/non-neuroendocrine cell fate. LCNECs are divided into two subgroups according to genomic alterations: type I LCNECs exhibit a neuroendocrine profile characterized by achaete-scute homolog 1 (ASCL1)high/delta-like protein 3 (DLL3)high/NOTCHlow and type II LCNECs show the pattern ASCL1low/DLL3low/NOTCHhigh. Here, we used immunohistochemistry, transmission electron microscopy, and digital analysis to examine the role of the Notch ligand DLL3 as an immunomarker of the neuroendocrine state and ASCL1 as a regulator of cell-cell interactions in SCLCs and LCNECs. High DLL3 and ASCL1 expression was associated with atypical submicroscopic characteristics involving nuclear size, chromatin arrangement, Golgi apparatus, and endoplasmic reticulum, and was characteristic of type I LCNECs with similarity to SCLCs, whereas low DLL3 and ASCL1 expression was found in both SCLCs and type II LCNECs. In patients diagnosed at an early stage who did not have metastasis and who underwent chemotherapy, DLL3high and ASCL1high SCLCs and type I LCNECs were associated with a better prognosis and a lower risk of death. The present findings suggested that DLL3/ASCL1 are potential therapeutic targets and prognostic indicators in patients with SCLCs or LCNECs.
A
bstract
Neutrinoless double-beta decay is a key process in particle physics. Its experimental investigation is the only viable method that can establish the Majorana nature of neutrinos, providing ...at the same time a sensitive inclusive test of lepton number violation. CROSS (Cryogenic Rare-event Observatory with Surface Sensitivity) aims at developing and testing a new bolometric technology to be applied to future large-scale experiments searching for neutrinoless double-beta decay of the promising nuclei
100
Mo and
130
Te. The limiting factor in large-scale bolometric searches for this rare process is the background induced by surface radioactive contamination, as shown by the results of the CUORE experiment. The basic concept of CROSS consists of rejecting this challenging background component by pulse-shape discrimination, assisted by a proper coating of the faces of the crystal containing the isotope of interest and serving as energy absorber of the bolometric detector. In this paper, we demonstrate that ultra-pure superconductive Al films deposited on the crystal surfaces act successfully as pulse-shape modifiers, both with fast and slow phonon sensors. Rejection factors higher than 99.9% of
α
surface radioactivity have been demonstrated in a series of prototypes based on crystals of Li
2
MoO
4
and TeO
2
. We have also shown that point-like energy depositions can be identified up to a distance of
∼
1 mm from the coated surface. The present program envisions an intermediate experiment to be installed underground in the Canfranc laboratory (Spain) in a CROSS-dedicated facility. This experiment, comprising
∼
3
×
10
25
nuclei of
100
Mo, will be a general test of the CROSS technology as well as a worldwide competitive search for neutrinoless double-beta decay, with sensitivity to the effective Majorana mass down to 70 meV in the most favorable conditions.
CUPID-Mo is a bolometric experiment to search for neutrinoless double-beta decay (
0
ν
β
β
) of
100
Mo
. In this article, we detail the CUPID-Mo detector concept, assembly and installation in the ...Modane underground laboratory, providing results from the first datasets. The CUPID-Mo detector consists of an array of 20
100
Mo
-enriched 0.2 kg
Li
2
MoO
4
crystals operated as scintillating bolometers at
∼
20
mK
. The
Li
2
MoO
4
crystals are complemented by 20 thin Ge optical bolometers to reject
α
events by the simultaneous detection of heat and scintillation light. We observe a good detector uniformity and an excellent energy resolution of 5.3 keV (6.5 keV) FWHM at 2615 keV, in calibration (physics) data. Light collection ensures the rejection of
α
particles at a level much higher than 99.9% – with equally high acceptance for
γ
/
β
events – in the region of interest for
100
Mo
0
ν
β
β
. We present limits on the crystals’ radiopurity:
≤
3
μ
Bq/kg
of
226
Ra
and
≤
2
μ
Bq/kg
of
232
Th
. We discuss the science reach of CUPID-Mo, which can set the most stringent half-life limit on the
100
Mo
0
ν
β
β
decay in half-a-year’s livetime. The achieved results show that CUPID-Mo is a successful demonstrator of the technology developed by the LUMINEU project and subsequently selected for the CUPID experiment, a proposed follow-up of CUORE, the currently running first tonne-scale bolometric
0
ν
β
β
experiment.
Dry Dilution Refrigerators (DDR) based on pulse tube cryo-coolers have started to replace Wet Dilution Refrigerators (WDR) due to the ease and low cost of operation. However these advantages come at ...the cost of increased vibrations, induced by the pulse tube. In this work, we present the vibration measurements performed on three different commercial DDRs. We describe in detail the vibration measurement system we assembled, based on commercial accelerometers, conditioner and DAQ, and examined the effects of the various damping solutions utilized on three different DDRs, both in the low and high frequency regions. Finally, we ran low temperature, pseudo-massive (30 and 250g) germanium bolometers in the best vibration-performing system under study and report on the results.
The common epidermal growth factor receptor (EGFR) mutations, such as the L858R point mutation in exon 21 and the in-frame deletional mutation in exon 19, have been definitively associated with ...response to EGFR-tyrosine kinase inhibitors (EGFR-TKI). However, the clinical outcome and response to treatment for many other rarer mutations are still unclear. In this study, we report the results of Brazilian patients in stage IB-IIIA non-small cell lung cancer (NSCLC) following complete resection with minimal residual disease and EGFR mutations treated with adjuvant chemotherapy and/or EGFR-TKIs. The frequency of EGFR mutations was investigated in 70 cases of early stage NSCLC. Mutations in exons 18 and 20, uncommon mutations in exons 19 and 21, as well as in exons 3, 7, 14, 16, 22, 27, and 28, and/or the presence of different mutations in a single tumor (complex mutations) are considered rare. EGFR mutations were detected in 23 tumors (32.9%). Fourteen cases carried rare mutations and were treated with platinum-based chemotherapy and two cases were treated with erlotinib. The clinical outcome is described case by case with references to the literature. Notably, we found two rare EGFR mutations and one of them with an unknown response to chemotherapy and/or EGFR-TKIs. We have provided complementary information concerning the clinical outcome and treatment of patients with early stage NSCLC for several rare EGFR mutations not previously or only rarely reported. Description of cases harboring rare mutations can support the decision-making process in this subset of patients.
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