Evidence for the involvement of lipid rafts localized at the ER-mitochondria associated membranes in autophagosome formation

Mitochondria-associated membranes (MAMs) are subdomains of the endoplasmic reticulum (ER) that interact with mitochondria. This membrane scrambling between ER and mitochondria appears to play a critical role in the earliest steps of autophagy. Recently, lipid microdomains, i.e. lipid rafts, have been identified as further actors of the autophagic process. In the present work, a series of biochemical and molecular analyses has been carried out in human fibroblasts with the specific aim of characterizing lipid rafts in MAMs and to decipher their possible implication in the autophagosome formation. In fact, the presence of lipid microdomains in MAMs has been detected and, in these structures, a molecular interaction of the ganglioside GD3, a paradigmatic “brick” of lipid rafts, with core-initiator proteins of autophagy, such as AMBRA1 and WIPI1, was revealed. This association seems thus to take place in the early phases of autophagic process in which MAMs have been hypothesized to play a key role. The functional activity of GD3 was suggested by the experiments carried out by knocking down ST8SIA1 gene expression, i.e., the synthase that leads to the ganglioside formation. This experimental condition results in fact in the impairment of the ER-mitochondria crosstalk and the subsequent hindering of autophagosome nucleation. We thus hypothesize that MAM raft-like microdomains could be pivotal in the initial organelle scrambling activity that finally leads to the formation of autophagosome. Introduction The interaction of the endoplasmic reticulum (ER) with mito- chondria occurs via certain subdomains of the ER, named mitochondria-associated membranes (MAMs), which allow membrane “scrambling” between these organelles and contrib- utes to the complex series of ER functions.1-3 Indeed, several regions of close apposition between the ER and mitochondria were detected by studies carried out several years ago.4,5 How- ever, since these studies provided only ultrastructural observa- tions, these reports remained neglected for a long time. In particular, while morphological evidence of the physical juxta- position between ER and mitochondria was described since 1959,6 it was experimentally proven only 30 y later. In fact, ana- lyzing ER fractions copurified with mitochondria in velocity sedimentation assays, mainly from rat liver cells, it was observed that mitochondria can tightly be associated with ele- ments of the ER and that the communication and intermixing between ER and mitochondria can be mediated by MAMs.7-12 These works also showed that these cosedimenting fractions were enriched in enzymes responsible for the synthesis of lipids. These findings suggested that MAMs could act as sites

The effect of radiation pressure on emission line profiles and black hole mass determination in active galactic nuclei

We present a new analysis of the motion of broad line region (BLR) clouds in active galactic nuclei (AGNs) taking into account the combined influence of gravity and radiation pressure force. We calculate cloud orbits under a large range of conditions and include the effect of a changing column density as a function of location. The dependence of radiation pressure force on the level of ionization and the column density are accurately computed. The main results are: a. The mean cloud location r(BLR) and the line widths (FWHMs) are combined in such a way that the simple virial mass estimate, r{BLR} FWHM^2/G, gives a reasonable approximation to the black hole mass M even when radiation pressure force is important. The reason is that L/M rather than L is the main parameter affecting the planar cloud motion. b. Reproducing the observed mean radius, FWHM and intensity of H-beta and CIV 1549 requires at least two different populations of clouds. c. The cloud location is a function of both L^{1/2} and L/M. Given this we suggest a new approximation for r(BLR) which, when inserted into the BH mass equation, results in a new approximation for M. The new expression involves L^{1/2}, FWHM and two constants that are obtained from a comparison with available M-sigma mass estimates. It deviates only slightly from the old mass estimate at all luminosities. d. The quality of present black hole mass estimators depends, critically, on the way the present M-sigma AGN sample (16 objects) represents the AGN population, in particular the distribution of Eddington ratios.Comment: Accepted for publication in ApJ, 11 pages, 9 figure

Apparent diffusion coefficient values in borderline breast lesions upgraded and not upgraded at definitive histopathological examination after surgical excision

Purpose: The study aims were to evaluate if the apparent diffusion coefficient (ADC) value could distinguish between breast lesions classified as B3 at core needle biopsy (CNB) that show or do not show atypia or malignancy at definitive histopathological examination (DHE) after surgical excision. Material and methods: From January 2013 to December 2017, 141 patients with a B3 breast lesion underwent magnetic resonance imaging and were included in the study. The ADC value was assessed drawing a ROI outlining the entire lesion, evaluating the mean ($ADC_{mean}$) and minimum ADC values ($ADC_{min}$). Results: Both $ADC_{mean}$ and ADCmin values showed a statistically significant difference between B3 lesions without and with malignancy or, for B3a lesions, atypia at DHE. They both showed a statistically significant difference also between B3a lesions without or with atypia or malignancy at DHE, but only $ADC_{min}$ (not $ADC_{mean}$) showed statistically significant difference between B3b lesions without or with malignancy at DHE. Conclusions: The ADC value could help distinguish between B3a lesions without or with atypia/malignancy at DHE after surgical excision and between B3b lesions without or with malignancy at DHE. Therefore, it could be used to help guide the diagnostic-therapeutic pathway of these lesions, particularly of B3a lesion

Does degradation from selective logging and illegal activities differently impact forest resources? A case study in Ghana

Degradation, a reduction of the ecosystem’s capacity to supply goods and services, is widespread in tropical forests and mainly caused by human disturbance. To maintain the full range of forest ecosystem services and support the development of effective conservation policies, we must understand the overall impact of degradation on different forest resources. This research investigates the response to disturbance of forest structure using several indicators: soil carbon content, arboreal richness and biodiversity, functional composition (guild and wood density), and productivity. We drew upon large field and remote sensing datasets from different forest types in Ghana, characterized by varied protection status, to investigate impacts of selective logging, and of illegal land use and resources extraction, which are the main disturbance causes in West Africa. Results indicate that functional composition and the overall number of species are less affected by degradation, while forest structure, soil carbon content and species abundance are seriously impacted, with resources distribution reflecting the protection level of the areas. Remote sensing analysis showed an increase in productivity in the last three decades, with higher resiliency to change in drier forest types, and stronger productivity correlation with solar radiation in the short dry season. The study region is affected by growing anthropogenic pressure on natural resources and by an increased climate variability: possible interactions of disturbance with climate are also discussed, together with the urgency to reduce degradation in order to preserve the full range of ecosystem functions

Omeprazole induces apoptosis in normal human polymorphonuclear leucocytes.

We investigated in vitro apoptosis in human polymorphonuclear neutrophils (PMN) induced by omeprazole. This drug, both in the native (OM) and acidified (OM-HCl) form, is a potent inducer of PMN apoptosis. The effect is time- and dose-dependent. OM-HCl is more efficient than OM in inducing PMN apoptosis. In fact, after 24 h incubation in vitro at 1×10 −4M OM-HCl induces apoptosis in 70% of the cell population compared to 37% induced by OM. Apoptosis induced by both forms of the drug is caspase dependent being significantly reduced by pretreating cells with the caspase 3 inhibitor (DEVDH-CHO). However, some differences in the apoptosis mechanisms between the two forms of the drug seem to exist because PMN treatment with the specific caspase 8 inhibitor (Z-IETD-FMK) only blocks OM-HCl mediated apoptosis. We observed cleavage of caspase 8 only in the cells incubated with OM-HCl while the executioner caspase 3 was activated with both forms of the drug. Furthermore, pretreatment with GM-CSF, a known activator of intracellular survival pathways in PMN, partially protected cells from OM-HCl induced apoptosis but did not contrast the apoptotic effect of OM. Cysteine cathepsin proteases also seem involved in the apoptotic mechanism of both drug forms since the specific inhibitor E64d gave a significant protection. To verify if OM-HCl induced apoptosis was dependent on the sulfenamide bound with the cell sulfhydryl groups we used molecules with thiol groups such as β-mercaptoethanol (β-ME) and reduced glutathione (GSH). Reactions of OM-HCl with cellular sulfhydryl groups are strongly involved in both the triggering and evolving phase of the apoptotic mechanism since significant protection from apoptosis was obtained when PMN were pretreated for 1h with β-ME (lipid-permeable) or GSH (lipid-impermeable). These results show that OM and OM-HCl induce apoptosis in human PMN and suggest that the second binds the sulfhydryl groups, present on the cell membrane, to then penetrate the cell thus causing a further significant increase in apoptosis. OM-induced PMN apoptosis during the treatment of gastric inflammatory disease could be an advantage for the resolution of the phlogosis state. However, this aspect should be further elucidated to assess the optimal therapeutical regimen for gastric diseases which are related to infective agents

The effect of radiation pressure on virial black hole mass estimates and the case of Narrow Line Seyfert 1 galaxies

We consider the effect of radiation pressure from ionizing photons on black hole (BH) mass estimates based on the application of the virial theorem to broad emission lines in AGN spectra. BH masses based only on the virial product V^2R and neglecting the effect of radiation pressure can be severely underestimated especially in objects close to the Eddington limit. We provide an empirical calibration of the correction for radiation pressure and we show that it is consistent with a simple physical model in which BLR clouds are optically thick to ionizing radiation and have average column densities of NH~10^23 cm^-2. This value is remarkably similar to what is required in standard BLR photoionization models to explain observed spectra. With the inclusion of radiation pressure the discrepancy between virial BH masses based on single epoch spectra and on reverberation mapping data drops from 0.4 to 0.2 dex rms. The use of single epoch observations as surrogates of reverberation mapping campaigns can thus provide more accurate BH masses than previously thought. Finally, we show that Narrow Line Seyfert 1 (NLS1) galaxies have apparently low BH masses because they are radiating close to their Eddington limit. After the radiation pressure correction, NLS1 galaxies have BH masses similar to other broad line AGNs and follow the same MBH-sigma/L relations as other active and normal galaxies. Radiation forces arising from ionizing photon momentum deposition constitute an important physical effect which must be taken into account when computing virial BH masses.Comment: To appear in the Astrophysical Journal, May 1 issu

Le parole sono ponti: risorse digitali per l'integrazione in contesti multilingui

Learning the language of the hosting country is a key factor for the integration of people coming from abroad but this objective could be achieved only if political institutions promote initiatives in this direction. Researchers in the field can cooperate, providing their expertise. In this paper we present two research activities concerning the use and the production of multilingual resources developed at the Institute for Computational Linguistics “Antonio Zampolli”. More specifically, we report on the creation of a multilingual glossary of difficult terms extracted from forms that people submit to the municipality of Genova to access public services. We also describe the experimental use in a primary school of ImagAct, a multilingual lexical resource focused on action verbs consulted by pupils to create a vocabulary of food preparation in Italian, English, Chinese and Spanish