Cholesterol reduction impairs exocytosis of synaptic vesicles

Cholesterol and sphingolipids are abundant in neuronal membranes, where they help the organisation of the membrane microdomains involved in major roles such as axonal and dendritic growth, and synapse and spine stability. The aim of this study was to analyse their roles in presynaptic physiology. We first confirmed the presence of proteins of the exocytic machinery (SNARES and Ca v 2.1 channels) in the lipid microdomains of cultured neurons, and then incubated the neurons with fumonisin B (an inhibitor of sphingolipid synthesis), or with mevastatin or zaragozic acid (two compounds that affect the synthesis of cholesterol by inhibiting HMG-CoA reductase or squalene synthase). The results demonstrate that fumonisin B and zaragozic acid efficiently decrease sphingolipid and cholesterol levels without greatly affecting the viability of neurons or the expression of synaptic proteins. Electron microscopy showed that the morphology and number of synaptic vesicles in the presynaptic boutons of cholesterol-depleted neurons were similar to those observed in control neurons. Zaragozic acid (but not fumonisin B) treatment impaired synaptic vesicle uptake of the lipophilic dye FM1-43 and an antibody directed against the luminal epitope of synaptotagmin-1, effects that depended on the reduction in cholesterol because they were reversed by cholesterol reloading. The time-lapse confocal imaging of neurons transfected with ecliptic SynaptopHluorin showed that cholesterol depletion affects the post-depolarisation increase in fluorescence intensity. Taken together, these findings show that reduced cholesterol levels impair synaptic vesicle exocytosis in cultured neurons

The Gribov problem in Noncommutative QED

It is shown that in the noncommutative version of QED (NCQED) Gribov copies induced by the noncommutativity of space-time appear in the Landau gauge. This is a genuine effect of noncommutative geometry which disappears when the noncommutative parameter vanishes.Comment: 19 pages, 3 figures. Published. The paper has been shortened and many references have been judged unnecessary or not suitable during the reviewing proces

Importin-beta and CRM1 control a RANBP2 spatiotemporal switch essential for mitotic kinetochore function

Protein conjugation with small ubiquitin-related modifier (SUMO) is a post-translational modification that modulates protein interactions and localisation. RANBP2 is a large nucleoporin endowed with SUMO E3 ligase and SUMO-stabilising activity, and is implicated in some cancer types. RANBP2 is part of a larger complex, consisting of SUMO-modified RANGAP1, the GTP-hydrolysis activating factor for the GTPase RAN. During mitosis, the RANBP2–SUMO-RANGAP1 complex localises to the mitotic spindle and to kinetochores after microtubule attachment. Here, we address the mechanisms that regulate this localisation and how they affect kinetochore functions. Using proximity ligation assays, we find that nuclear transport receptors importin-β and CRM1 play essential roles in localising the RANBP2–SUMO-RANGAP1 complex away from, or at kinetochores, respectively. Using newly generated inducible cell lines, we show that overexpression of nuclear transport receptors affects the timing of RANBP2 localisation in opposite ways. Concomitantly, kinetochore functions are also affected, including the accumulation of SUMO- conjugated topoisomerase-IIα and stability of kinetochore fibres. These results delineate a novel mechanism through which nuclear transport receptors govern the functional state of kinetochores by regulating the timely deposition of RANBP2

N-type Ca2+ Channels Are Present in Secretory Granules and Are Transiently Translocated to the Plasma Membrane during Regulated Exocytosis

Abstract An intracellular pool of N-type voltage-operated calcium channels has recently been described in different neuronal cell lines. We have now further characterized the intracellular pool of N-type calcium channels in both IMR32 human neuroblastoma and PC12 rat pheochromocytoma cells. Intracellular N-type calcium channels were found to be accumulated in subcellular fractions where the chromogranin B-containing secretory granules were also enriched. 125I-ω-Conotoxin GVIA binding assays on fixed and permeabilized cells revealed that intracellular N-type calcium channels translocate to the plasma membrane in cells exposed to secretagogues (KCl, ionomycin, and phorbol esters). The kinetics, Ca2+ and protein kinase C dependence, and brefeldin A insensitivity of N-type calcium channels translocation were similar to the regulated release of chromogranin B, while no correlation was found with the constitutive secretion of a heparan sulfate proteoglycan. A PC12 subclone deficient in the regulated but not in the constitutive pathway of secretion had a small intracellular pool of N-type calcium channels, and no secretagogueinduced translocation occurred in these cells. Calcium channel translocation was accompanied by a stronger response of Fura-2-loaded cells to depolarizing stimuli, suggesting that the newly inserted channels are functional

Stress echocardiography for the risk stratification of patients following coronary bypass surgery

Objectives: The aim of the study was to assess the prognostic value of stress echocardiography after surgical revascularization. Methods: We evaluated 500 (100 women) patients who had undergone exercise or pharmacological SE after a median of 69 months after coronary artery by-pass grafting (CABG). Of these, 351 (70%) complained of symptoms suggestive of ischemic origin while 149 (30%) were tested for asymptomatic progression of the disease. Results: SE was positive for ischemia in 196 (39%) patients. During a median follow-up of 25 months, 61 patients died, 33 had a nonfatal myocardial infarction, and 112 underwent late (N3 months) revascularization. Multivariable Cox\u27 regression analysis indicated age (HR=1.04; 95% CI 1.01-1.06; pb0.003), and peak WMSI (HR=3.07; 95% CI 1.96-4.81; p=0.0001) as independent predictors of hard (total mortality and myocardial infarction) events. SE information provided a significant improvement in predictive power of the statistical model (chi-square increase 34%, pb0.0001 for hard and 91%, pb0.0001 for major events, respectively). Survival analysis showed ischemia at SE to be associated with significantly higher hard and major event rate in both symptomatic and asymptomatic patients. Discussion: SE represents an effective tool for the risk stratification of patients with previous CABG independently of the presence of symptoms suggestive of ischemic origin

Hepatitis E virus in Italy: molecular analysis of travel-related and autochthonous cases

Human hepatitis E virus (HEV) is considered an emerging pathogen in industrialized countries. The aim of the present study was to contribute to the body of knowledge available on the molecular epidemiology of acute hepatitis E in Italy. Three sets of HEV-specific primers targeting the ORF1 and ORF2 were used to examine serum samples collected from acute hepatitis patients positive for anti-HEV IgG and/or IgM, between 2007 and 2010. Seventeen patients (39.5 %) tested HEV RNA-positive: 12 infections, due to genotype 1, were associated with travel to endemic areas (Bangladesh, India and Pakistan), while five infections, due to genotype 3, were presumably autochthonous. Risk factors identified in this group included exposure to raw seafood, pork liver sausages and wild boar. Results from the present study confirm that human HEV infection in Italy is caused by different genotypes, depending on whether the infection is travel-related or autochthonous

Strategies and Techniques for Powering Wireless Sensor Nodes through Energy Harvesting and Wireless Power Transfer

The continuous development of the internet of things (IoT) infrastructure and applications is paving the way for advanced and innovative ideas and solutions, some of which are pushing the limits of state-of-the-art technology. The increasing demand for Wireless Sensor Nodes (WSNs) able to collect and transmit data through wireless communication channels, while often positioned in locations that are difficult to access, is driving research into innovative solutions involving energy harvesting (EH) and wireless power transfer (WPT) to eventually allow battery-free sensor nodes. Due to the pervasiveness of radio frequency (RF) energy, RF EH and WPT are key technologies with the potential to power IoT devices and smart sensing architectures involving nodes that need to be wireless, maintenance free, and sufficiently low in cost to promote their use almost anywhere. This paper presents a state-of-the-art, ultra-low power 2.5 W highly integrated mixed-signal system on chip (SoC), for multi-source energy harvesting and wireless power transfer. It introduces a novel architecture that integrates an ultra-low power intelligent power management, an RF to DC converter with very low power sensitivity and high power conversion efficiency (PCE), an Amplitude-Shift-Keying/Frequency-Shift-Keying (ASK/FSK) receiver and digital circuitry to achieve the advantage to cope, in a versatile way and with minimal use of external components, with the wide variety of energy sources and use cases. Diverse methods for powering wireless Sensor Nodes through energy harvesting and wireless power transfer are implemented providing related system architectures and experimental results

Unusual Methylobacterium fujisawaense Infection in a Patient with Acute Leukaemia Undergoing Hematopoietic Stem Cell Transplantation: First Case Report

Microorganisms of the genus Methylobacterium are facultative methylotrophic, gram-negative rods that are ubiquitous in nature and rarely cause human disease, mostly in subjects with preexisting causes of immune depression. Methylobacterium fujisawaense, first proposed as a new species in 1988, has never been reported as a bacterial agent of human infections so far. Here we describe a case of M. fujisawaense infection in a relapsed acute leukaemia undergoing unrelated allogeneic hematopoietic stem cell transplantation. Molecular identification of an M. fujisawaense strain was obtained from multiple mycobacterial blood cultures