Reactive oxygen species plasmatic levels in ischemic stroke

Oxidative stress is probably one of the mechanisms involved in neuronal damage induced by ischemia-reperfusion, and the antioxidant activity of plasma may be an important factor providing protection from neurological damage caused by stroke-associated oxidative stress. The aim of this study was to investigate the status of oxidative stress, NO and ONOO⁻ levels in patients with atherothrombotic and lacunar acute ischemic stroke and iNOS, eNOS and nitrotyrosine expression in the same patients. Plasma ONOO⁻ levels were significantly higher in patients than in controls while NO decreases in patients in respect to controls. Densitometric analysis of bands indicated that iNOS and N-Tyr protein levels were significantly higher in patients in respect to controls. This study has highlighted a significant NO decrease in our patients compared with controls and this is most probably due to the increased expression of inducible NO synthase by the effect of thrombotic attack. In fact, the constitutive NO isoforms, which produce small amounts of NO, are beneficial, while activation of the inducible isoform of NO, which produces much more NO, causes injury, being its toxicity greatly enhanced by generation of peroxynitrite. The significant ONOO⁻ increase observed in our patients, compared to controls, is most probably due to reaction of NO with O₂^·−. These findings suggest that free radical production and oxidative stress in ischemic stroke might have a major role in the pathogenesis of ischemic brain injury. Peroxynitrite might be the main marker of brain damage and neurological impairment in acute ischemic stroke.     

Epigenetic control of the critical region for premature ovarian failure on autosomal genes translocated to the X chromosome: a hypothesis

Chromosomal rearrangements in Xq are frequently associated to premature ovarian failure (POF) and have contributed to define a POF “critical region” from Xq13.3 to Xq26. Search for X-linked genes responsible for the phenotype has been elusive as most rearrangements did not interrupt genes and many were mapped to gene deserts. We now report that ovary-expressed genes flanked autosomal breakpoints in four POF cases analyzed whose X chromosome breakpoints interrupted a gene poor region in Xq21, where no ovary-expressed candidate genes could be found. We also show that the global down regulation in the oocyte and up regulation in the ovary of X-linked genes compared to the autosomes is mainly due to genes in the POF “critical region”. We thus propose that POF, in X;autosome balanced translocations, may not only be caused by haploinsufficiency, but also by a oocyte-specific position effect on autosomal genes, dependent on dosage compensation mechanisms operating on the active X chromosome in mammals.     

Spreeta-based biosensor assays for endocrine disruptors

The construction and performance of an automated low-cost Spreeta-based prototype biosensor system for the detection of endocrine disrupting chemicals (EDCs) is described. The system consists primarily of a Spreeta miniature liquid sensor incorporated into an aluminum flow cell holder, dedicated to support a Biacore chip frame, in combination with a simple pressurized air-driven fluid system. During the optimization, a monoclonal antibody (MAb)-based immunoassay for the estrogenic compound bisphenol A (BPA) was used as a model. After the optimization two thyroxine transport protein inhibition assays for thyroid endocrine disruptors were implemented. The average noise of the system for 1 min of baseline was 1.1 microRIU (refractive index units) and it could be operated in the range of 18-22 degrees C with a minimum baseline drift (5-10 microRIU/100 min). Optimum signal to noise ratio (S/N R) was obtained using a flow cell height of 100 microm and a flow rate of 180 microl/min. The sensitivity of the Spreeta-based biosensor inhibition assays implemented (50% inhibition concentration (IC50) of 30.2 nM for BPA using MAb12 and 12.3 and 11.6 nM for thyroxine (T4) using thyroxine-binding globulin (TBG) and recombinant transthyretin (rTTR), respectively) was comparable to the sensitivity previously obtained using a Biacore 3000 (IC50 of 39.9 nM for BPA and 8.6 and 13.7 nM, respectively, for T4). The results indicate that the alternative prototype system can be used in combination with ready-to-use biosensor chip surfaces and it is potentially a useful tool for the bioeffect-related screening of EDCs.     

Is the Eco-label EU Decision for hard coverings really capable of capturing the environmental performances of the marble productive chain? A field verification by means of a life cycle approach

Purpose

This work intends to show whether the Eco-label EU Decision for hard coverings, in which marble is contemplated among hard coverings as a natural product, is really capable of capturing the environmental performances of the marble productive chain, in other words whether it is actually viable for the natural products, like marble.

Methods

After a preliminary critical analysis of the suitability for marble of the current EU Decision (2009/607/EC), a classical life cycle assessment (LCA) methodology has been applied on the field to the marble “Perlato di Sicilia.” More specifically, the whole productive chain of a couple of firms treating the “Perlato di Sicilia” marble has been examined. The life cycle analysis is actually a cradle-to-gate analysis which includes the raw material extraction, processing phase, and finishing operations.

Results and discussion

Both the preliminary critical analyses of the structure of the Decision and the in-field checking on the two firms of the Custonaci marble district in Sicily singled out several conflicting points of the Decision for the marble working chain. These difficulties could be reasonably extended to other natural stones, such as granite, for which similar working processes are applied. Based on the outcomes of both these analyses, in the present work, a set of new indicators and modified criteria, already present in the Decision, is proposed as a candidate to be considered for inclusion in a future release of the Decision.

Conclusions

The changes here introduced can represent a useful indication toward a more suitable scheme of the EU Eco-label for marble, at least. Clearly, further investigations need to better assess the proposed scheme, especially in terms of threshold values of pollutant releases and use of explosive that are actually specific for the marble productive chain. The present modified version of the standard has been proposed to the Sicilian administration in order to be voluntarily adopted by marble productive sites of the region, in the aim of extensively verifying its suitability.     

Different functions of HOPS isoforms in the cell

Hepatocyte odd protein shuttling (HOPS) moves between nucleus and cytoplasm. HOPS overexpression leads to cell cycle arrest in G₀/G₁, and HOPS knockdown causes centrosome alterations, with subsequent abnormal cell division. Recently, we demonstrated that HOPS acts as a functional bridge in NPM-p19^Arf interactions. Here we show that HOPS is present in 3 different isoforms that play distinct intracellular functions. Although HOPS is a transmembrane ubiquitin, an isoform with intermediate molecular weight is cleaved from the membrane and released into the cytosol, to act as the shuttling protein. We identified a signal peptide peptidase structure in N-terminal membrane-bound HOPS that allows the regulated intramembrane proteolysis (RIP) system to control the relative amounts of the released, shuttling isoform capable of binding NPM. These results argue for distinct, isoform-specific functions of HOPS in the nucleolus, nucleus, and cytoplasm and provide insight into the dynamics of HOPS association with NPM, whose mutation and subsequent delocalization is found in 30% of acute myeloid leukemia patients.