Role of Protein Ubiquitination in NFH-LacZ mice

In neurodegenerative diseases, one can observe deposits of degradation products that represent hallmark structures. Actually, the underlying mechanisms are not well understood, but some hypotheses claim that the ubiquitin-proteasome system is perturbed in neurodegenerative diseases. Some of the influencing factors are aging, oxidation and the formation of free radicals, as well as genetic mutations which affect the function of proteins and result in an accumulation and formation of aggresomes. The amyotrophic lateral sclerosis, in which a malfunction of the sodium dismutase perturbs the redox system, is characterized by the accumulation of elements of the cytoskeleton in motor neurons and a progressive neuronal death. We suppose that in these diseases the ubiquitin- proteasome system is deregulated and try to demonstrate this hypothesis by comparing the ubiquitination of different neurofilaments in brain and spinal cord of transgenic and control mice. These NFH-LacZ mice with a truncated NF-H protein and a ß-galactosidase marker protein induce an accumulation of NF-proteins and neurofilaments are no longer transported into axons or dendrites. The accumulation of such aggregates resembles the phenotype of amyotrophic lateral sclerosis. Beside the ubiquitination the neurofilament expression and phosphorylation state was investigated. The results cannot demonstrate a perturbation of the ubiquitin-proteasome system of neurofilaments in transgenic mice. In contrast, in accordance with the mechanism of the NFH-LacZ mice a decrease of high and medium density neurofilaments and a hypophosphorylation were found. In conclusion, to elicit the pathological mechanism of amyotrophic lateral sclerosis and to develop focused treatments, we have to review the pathological mechanism of the transgenic mice and repeat the experiments with other animal models or with human material. Other possibilities would be to focus on other degradation mechanisms, such as the endosome/lysosome system, and to define their role in the amyotrophic lateral sclerosis more clearly

Cyclic voltammetry : a tool to quantify 2,4,6-trichloroanisole in aqueous samples from cork planks boiling industrial process

Chloroanisoles, namely 2,4,6-trichloroanisole, are pointed out as the primary responsible of the development of musty off-flavours in bottled wine, due to their migration from cork stoppers, which results in huge economical losses for wine industry. A prevention step is the detection of these compounds in cork planks before stoppers are produced. Mass spectrometry gas chromatography is the reference method used although it is far beyond economical possibilities of the majority of cork stoppers producers. In this work, a portable cyclic voltammetry approach was used to detect 2,4,6-trichloroanisole extracted from natural cork planks to the aqueous phase during the cork boiling industrial treatment process. Analyses were carried out under ambient conditions, in less than 15 min with a low use of solvent and without any sample pre-treatment. The proposed technique had detection (0.31±0.01 ng/L) and quantification (0.95±0.05 ng/L) limits lower than the human threshold detection level. For blank solutions, without 2,4,6-trichloroanisole addition, a concentration in the order of the quantification limit was estimated (1.0±0.2 ng/L), which confirms the satisfactory performance of the proposed methodology. For aqueous samples from the industrial cork planks boiling procedure, intra-day repeatabilities were lower than 3%, respectively. Also, 2,4,6-trichloroanisole contents in the aqueous samples determined by this novel approach were in good agreement with those obtained by GC-MS (correlation coefficient equal to 0.98), confirming the satisfactory accuracy of the proposed methodology. So, since this novel approach is a fast, low-cost, portable and user-friendly method, it can be an alternative and helpful tool for in-situ industrial applications, allowing accurate detection of releasable 2,4,6-trichloroanisole in an earlier phase of cork stoppers production, which may allow implementing more effective cork treatments to reduce or avoid future 2,4,6-trichloroanisole contaminations of wine.This work was partially supported by project PEst-C/EQB/LA0020/2011, financed by FEDER through COMPETE - Programa Operacional Factores de Competitividade and by FCT - Fundacao para a Ciencia e a Tecnologia (Portugal)

Toward Unraveling Grape Tannin Composition: Application of Online Hydrophilic Interaction Chromatography × Reversed-Phase Liquid Chromatography–Time-of-Flight Mass Spectrometry for Grape Seed Analysis

Despite the significant importance of tannins in viticulture and enology, relatively little is known about the detailed chemical composition of these molecules. This is due to challenges associated with the accurate analytical determination of the highly structurally diverse proanthocyanidins which comprise tannins. In this contribution, we address this limitation by demonstrating how online comprehensive two-dimensional liquid chromatography (LC × LC) coupled to high resolution mass spectrometry (HR-MS) can be exploited as a powerful analytical approach for the detailed characterization of grape seed tannins. Hydrophilic interaction chromatography (HILIC) and reversed-phase liquid chromatography (RP-LC) were employed in the two dimensions to provide complementary information in terms of separation according to hydrophilicity and hydrophobicity, respectively. Online coupling of HILIC × RP-LC with fluorescence detection and electrospray ionization MS delivered high resolution analysis in a practical analysis time, while allowing selective detection and facilitating compound identification. Time-of-flight (TOF) MS provided high acquisition rates and sensitivity coupled to accurate mass information, which allowed detection of procyanidins up to a degree of polymerization (DP) of 16 and a degree of galloylation up to 8 in a red grape seed extract. This analytical methodology promises to shed new light on these important grape constituents and potentially on their evolution during wine production

Investigation of the volatile composition of pinotage wines fermented with different malolactic starter cultures using comprehensive two-dimensional gas chromatography coupled to time-of-flight mass spectrometry (GC×GC-TOF-MS)

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Shape Correspondence with Isometric and Non-Isometric Deformations

The registration of surfaces with non-rigid deformation, especially non-isometric deformations, is a challenging problem. When applying such techniques to real scans, the problem is compounded by topological and geometric inconsistencies between shapes. In this paper, we capture a benchmark dataset of scanned 3D shapes undergoing various controlled deformations (articulating, bending, stretching and topologically changing), along with ground truth correspondences. With the aid of this tiered benchmark of increasingly challenging real scans, we explore this problem and investigate how robust current state-of- the-art methods perform in different challenging registration and correspondence scenarios. We discover that changes in topology is a challenging problem for some methods and that machine learning-based approaches prove to be more capable of handling non-isometric deformations on shapes that are moderately similar to the training set