Quantitative Characterization of α-Synuclein Aggregation in Living Cells through Automated Microfluidics Feedback Control

Aggregation of α-synuclein and formation of inclusions are hallmarks of Parkinson's disease (PD). Aggregate formation is affected by cellular environment, but it has been studied almost exclusively in cell-free systems. We quantitatively analyzed α-synuclein inclusion formation and clearance in a yeast cell model of PD expressing either wild-type (WT) α-synuclein or the disease-associated A53T mutant from the galactose (Gal)-inducible promoter. A computer-controlled microfluidics device regulated α-synuclein in cells by means of closed-loop feedback control. We demonstrated that inclusion formation is strictly concentration dependent and that the aggregation threshold of the A53T mutant is about half of the WT α-synuclein (56%). We chemically modulated the proteasomal and autophagic pathways and demonstrated that autophagy is the main determinant of A53T α-synuclein inclusions’ clearance. In addition to proposing a technology to overcome current limitations in dynamically regulating protein expression levels, our results contribute to the biology of PD and have relevance for therapeutic applications

Cervical Laminoplasty with Polypropylene Mesh: Our Experience

Cervical laminoplasty was made to reduce the complications of the simple laminectomy. The main indication is degenerative cervical myelopathy with posterior compression to two or more levels. To simplify this procedure with more effectiveness, we conceived a cervical laminoplastica with polypropylene mesh, generally used in abdominoplasty, to provide more resistance during mechanisms of flexion-extension. MATERIALS AND METHODS: We analyzed 35 patients (27 males, 8 females) with degenerative cervical myelopathy who underwent surgical tretament at our institution between 2010 and 2015. RESULTS AND DISCUSSION: The main duration of surgery was about 90 minutes without intra or post-operative complications. Clinical and radiological follow-up was done at 1, 3 and 6 months, with cervical dynamic Rx and cervical RM at 1one year. There were no complications and 70% of the patients experienced a stable clinical improvement, while in 30% of patients stabilization of clinical deficits. The duration of the superior clinical to one year and the age gt of 70 years the more meaningful negative prognostic indicators have appeared. CONCLUSIONS: A clinical history more than one year and the age elder than 70 years appear the main negative prognostic factors. The advantage of the laminoplasty is a smaller anatomical damage of the posterior elements without cervical instability, and use of polypropylene mesh provides more resistance during flexion-extension

Effects of balloon injury on neointimal hyperplasia in steptozotocin-induced diabetes and in hyperinsulinemic nondiabetic pancreatic islet-transplanted rats.

BACKGROUND: The mechanisms of increased neointimal hyperplasia after coronary interventions in diabetic patients are still unknown. METHODS AND RESULTS: Glucose and insulin effects on in vitro vascular smooth muscle cell (VSMC) proliferation and migration were assessed. The effect of balloon injury on neointimal hyperplasia was studied in streptozotocin-induced diabetic rats with or without adjunct insulin therapy. To study the effect of balloon injury in nondiabetic rats with hyperinsulinemia, pancreatic islets were transplanted under the kidney capsule in normal rats. Glucose did not increase VSMC proliferation and migration in vitro. In contrast, insulin induced a significant increase in VSMC proliferation and migration in cell cultures. Furthermore, in VSMC culture, insulin increased MAPK activation. A reduction in neointimal hyperplasia was consistently documented after vascular injury in hyperglycemic streptozotocin-induced diabetic rats. Insulin therapy significantly increased neointimal hyperplasia in these rats. This effect of hyperinsulinemia was totally abolished by transfection on the arterial wall of the N17H-ras-negative mutant gene. Finally, after experimental balloon angioplasty in hyperinsulinemic nondiabetic islet-transplanted rats, a significant increase in neointimal hyperplasia was observed. CONCLUSIONS: In rats with streptozotocin-induced diabetes, balloon injury was not associated with an increase in neointimal formation. Exogenous insulin administration in diabetic rats and islet transplantation in nondiabetic rats increased both blood insulin levels and neointimal hyperplasia after balloon injury. Hyperinsulinemia through activation of the ras/MAPK pathway, rather than hyperglycemia per se, seems to be of crucial importance in determining the exaggerated neointimal hyperplasia after balloon angioplasty in diabetic animals

Deterministic Annealing as a jet clustering algorithm in hadronic collisions

We show that a general purpose clusterization algorithm, Deterministic Annealing, can be adapted to the problem of jet identification in particle production by high energy collisions. In particular we consider the problem of jet searching in events generated at hadronic colliders. Deterministic Annealing is able to reproduce the results obtained by traditional jet algorithms and to exhibit a higher degree of flexibility.Comment: 13 pages, 6 figure

Study of timing characteristics of a 3 m long plastic scintillator counter using waveform digitizers

A plastic scintillator bar with dimensions 300 cm x 2.5 cm x 11 cm was exposed to a focused muon beam to study its light yield and timing characteristics as a function of position and angle of incidence. The scintillating light was read out at both ends by photomultiplier tubes whose pulse shapes were recorded by waveform digitizers. Results obtained with the WAVECATCHER and SAMPIC digitizers are analyzed and compared. A discussion of the various factors affecting the timing resolution is presented. Prospects for applications of plastic scintillator technology in large-scale particle physics detectors with timing resolution around 100 ps are provided in light of the results

Automatic synchronisation of the cell cycle in budding yeast through closed-loop feedback control

The cell cycle is the process by which eukaryotic cells replicate. Yeast cells cycle asynchronously with each cell in the population budding at a different time. Although there are several experimental approaches to synchronise cells, these usually work only in the short-term. Here, we build a cyber-genetic system to achieve long-term synchronisation of the cell population, by interfacing genetically modified yeast cells with a computer by means of microfluidics to dynamically change medium, and a microscope to estimate cell cycle phases of individual cells. The computer implements a controller algorithm to decide when, and for how long, to change the growth medium to synchronise the cell-cycle across the population. Our work builds upon solid theoretical foundations provided by Control Engineering. In addition to providing an avenue for yeast cell cycle synchronisation, our work shows that control engineering can be used to automatically steer complex biological processes towards desired behaviours similarly to what is currently done with robots and autonomous vehicles

Ageing, sex and cardioprotection

Translation of cardioprotective interventions aimed at reducing myocardial injury during ischaemia-reperfusion from experimental studies to clinical practice is an important yet unmet need in cardiovascular medicine. One particular challenge facing translation is the existence of demographic and clinical factors that influence the pathophysiology of ischaemia-reperfusion injury of the heart and the effects of treatments aimed at preventing it. Among these factors, age and sex are prominent and have a recognised role in the susceptibility and outcome of ischaemic heart disease. Remarkably, some of the most powerful cardioprotective strategies proven to be effective in young animals become ineffective during ageing. This article reviews the mechanisms and implications of the modulatory effects of ageing and sex on myocardial ischaemia-reperfusion injury and their potential effects on cardioprotective interventions

VHMPID: a new detector for the ALICE experiment at LHC

This article presents the basic idea of VHMPID, an upgrade detector for the ALICE experiment at LHC, CERN. The main goal of this detector is to extend the particle identification capabilities of ALICE to give more insight into the evolution of the hot and dense matter created in Pb-Pb collisions. Starting from the physics motivations and working principles the challenges and current status of development is detailed.Comment: 4 pages, 6 figures. To be published in EPJ Web of Conference

Contribution of the HMPID detector to the high-pT physics at LHC

The LHC will deliver unexplored energy regimes for proton-proton and heavy-ion collisions. As shown by the RHIC experiments, particle identification over a large momentum range is essential to disentangle physics processes, especially in the intermediate p$_T$ (1 $<p_{T}<5$ GeV/c) region. The novel design of the High-Momentum Particle Identification Detector (HMPID), based on large surface CsI photocathodes, is able to identify $\pi^{\pm}$, $K^{\pm}$, $p$ and $\bar{p}$ in the momentum region where bulk medium properties and hard scatterings interplay. Furthermore, measurement of resonance particles such as the $\phi \to K^+K^-$ could provide information on the system evolution. The HMPID layout and segmentation are optimized to study particle correlations at high momenta describing the early phase and the dynamical evolution of the collision. At LHC, the increased hard cross section will significantly be enhanced compared to RHIC. Jet reconstruction via Deterministic Annealing can address jet quenching and detailed measurements of jet properties. In this paper, we present these selected topics from the possible HMPID contributions to the physics goals of LHC.Comment: 6 pages, 7 figures, Contribution to QCD @ Work 2007: International Workshop on Quantum Chromodynamics Theory and Experiment, Martina Franca, Italy, 16-20 June 200