Distribution of nitric oxide synthase immunoreactivity in the mouse brain

Nitric oxide (NO) is a gaseous intercellular messenger with a wide range of neural functions. NO is synthesized by activation of different isoforms of nitric oxide synthases (NOS). At present NOS immunoreactivity has been described in mouse brain in restricted and definite areas and no detailed mapping studies have yet been reported for NOS immunoreactivity. We have studied the distribution of neuronal NOS-containing neurons in the brain of three months male mice, using a specific commercial polyclonal antibody against the neuronal isoform of nitric oxide synthase (nNOS). Neuronal cell bodies exhibiting nNOS immunoreactivity were found in several distinct nuclei throughout the brain. The neurons that were positively stained exhibited different intensities of reaction. In some brain areas (i.e., cortex, striatum, tegmental nuclei) neurons were intensely stained in a Golgi-like fashion. In other regions, immunoreactive cells are moderately stained (i.e., magnocellular nucleus of the posterior commissure, amygdaloid nucleus, interpeduncular nucleus, lateral periaqueductal gray) or weakly stained (i.e., vascular organ of the lamina terminalis, hippocampus, inferior colliculus, reticular nucleus). In the mouse, the NO-producing system appears well developed and widely diffused. In particular, nNOS immunoreactive neurons seem chiefly present in several sensory pathways like all the nuclei of the olfactory system, as well as in many regions of the lymbic system. These data suggest a widespread role for the NO system in the mouse nervous system.     

Mass spectrometry for detection of 4-hydroxy-trans-2-nonenal (HNE) adducts with peptides and proteins

Despite the great technical advancement of mass spectrometry, this technique has contributed in a limited way to the discovery and quantitation of specific/precocious markers linked to free radical-mediated diseases. Unsaturated aldehydes generated by free radical-induced lipid peroxidation of polyunsaturated fatty acids, and in particular 4-hydroxy-trans-2 nonenal (HNE), are involved in the onset and progression of many pathologies such as cardiovascular (atherosclerosis, long-term complications of diabetes) and neurodegenerative diseases (Alzheimer's disease, Parkinson's disease, and cerebral ischemia). Most of the biological effects of HNE are attributed to the capacity of HNE to react with the nucleophilic sites of proteins and peptides (other than nucleic acids), to form covalently modified biomolecules that can disrupt important cellular functions and induce mutations. By considering the emerging role of HNE in several human diseases, an unequivocal analytical approach as mass spectrometry to detect/elucidate the structure of protein-HNE adducts in biological matrices is strictly needed not only to understand the reaction mechanism of HNE, but also to gain a deeper insight into the pathological role of HNE. This with the aim to provide intermediate diagnostic biomarkers for human diseases. This review sheds focus on the "state-of-the-art" of mass spectrometric applications in the field of HNE-protein adducts characterization, starting from the fundamental early studies and discussing the different MS-based approaches that can provide detailed information on the mechanistic aspects of HNE-protein interaction. In the last decade, the increases in the accessible mass ranges of modern instruments and advances in ionization methods have made possible a fundamental improvement in the analysis of protein-HNE adducts by mass spectrometry, and in particular by matrix-assisted laser desorption/ionization (MALDI) and electrospray ionization (ESI) tandem mass spectrometry. The recent developments and uses of combined analytical approaches to detect and characterize the type/site of interaction have been highlighted, and several other aspects, including sample preparation methodologies, structure elucidation, and data analysis have also been considered.     

Determination of 210pb and 212pb in water and their radiological impact to the public via drinking water

A sensitive and accurate method for determining ultra low-level 210Pb and 212Pb in water samples through double measurements was developed.Pb was pre-concentrated as hydroxides,separated from alkaline earth elements as PbS precipitate,purified by an anion exchange resin chromatography column,precipitated as PbSO4 for source preparation and counted by a low background β-counter.The procedure was checked with IAEA reference materials,and the results agreed well with the recommended values.Theminimum detectable activity was 0.062 mBq·L-1 for 210Pb and 0.053 mBq·L-1 for 212Pb with a 48 L water sample.Seventeen drinking water samples were analyzed,with a Pb recovery of 88.8?5.5%,and the typical activity concentrations were 0.191?15.1 mBq·L-1 for 210Pb and of 1.12?5.77 mBq·L-1 for 212Pb.The estimated committed effective doses to adult members of the public in Italy due to intake of 210Pb and 212Pb in drinking water were 0.096?7.59 μSv·a-1 and 0.005?0.025 μSv·a-1,respectively.     

From first principles to the Burrows and Wheeler transform and beyond,via combinatorial optimization

We introduce a combinatorial optimization framework that naturally induces a class of optimal word permutations with respect to a suitably defined cost function taking into account various measures of relatedness between words. The Burrows and Wheeler transform (bwt) (cf. [M. Burrows, D. Wheeler, A block sorting lossless data compression algorithm, Technical Report 124, Digital Equipment Corporation, 1994]), and its analog for labelled trees (cf. [P. Ferragina, F. Luccio, G. Manzini, S. Muthukrishnan, Structuring labeled trees for optimal succinctness, and beyond, in: Proc. of the 45th Annual IEEE Symposium on Foundations of Computer Science, 2005, pp. 198–207]), are special cases in the class. We also show that the class of optimal word permutations defined here is identical to the one identified by Ferragina et al. for compression boosting [P. Ferragina, R. Giancarlo, G. Manzini, M. Sciortino, Boosting textual compression in optimal linear time, Journal of the ACM 52 (2005) 688–713]. Therefore, they are all highly compressible. We also provide, by using techniques from Combinatorics on Words, a fast method to compute bwt without using any end-of-string symbol. We also investigate more general classes of optimal word permutations, where relatedness of symbols may be measured by functions more complex than context length. For this general problem we provide an instance that is MAX-SNP hard, and therefore unlikely to be solved or approximated efficiently. The results presented here indicate that a key feature of the Burrows and Wheeler transform seems to be, besides compressibility, the existence of efficient algorithms for its computation and inversion.     

Online Portable Microcantilever Biosensors for Salmonella enterica Serotype Enteritidis Detection

The micro- and nano-technologies coupled with a deep knowledge of organic/inorganic interfaces guarantee an exceptional sensitivity and specificity of the sensor, while the lab-on-a-chip platform reduces assay times and limits sampling and/or sample preparation, providing compact and portable objects. Therefore, the development of innovative biosensors such as antibody-immobilized microcantilevers can overcome the evident limits of nowadays technologies, such as time consuming, expensiveness, difficult automation, low sensitivity, accuracy, and precision for quantitative methods. The present study proposes two device designs for the detection of food pathogens, exploiting an antibody-immobilized microcantilever biosensors, a novel class of mass detectors. For the first one, we integrated the mechanical sensors on a microfluidic platform (lab-on-a-chip) to perform online analysis, directly in liquid environment. We showed that our portable biosensors could easily detect the presence of pathogenic bacteria such as Salmonella enterica serotype enteritidis in concentration 10⁵ cfu/mL in just 40 min, without any enrichment and/or sample preparation. To increase the mass sensitivity of our analysis, we also fabricated microstructures optimized for vibrating in vacuum environment. Using a dip-and-dry technique, we showed that, in such configuration, the experimental limit of detection is as low as 10³ cfu/mL. Due to the extremely small volumes needed, our biosensors operating in vacuum have the potentiality of detecting the presence or absence of a single cell.     

Immobilization of Lathyrus cicera Amine Oxidase on Magnetic Microparticles for Biocatalytic Applications

Amine oxidases are enzymes belonging to the class of oxidoreductases that are widespread, from bacteria to humans. The amine oxidase from Lathyrus cicera has recently appeared in the landscape of biocatalysis, showing good potential in the green synthesis of aldehydes. This enzyme catalyzes the oxidative deamination of a wide range of primary amines into the corresponding aldehydes but its use as a biocatalyst is challenging due to the possible inactivation that might occur at high product concentrations. Here, we show that the enzyme’s performance can be greatly improved by immobilization on solid supports. The best results are achieved using amino-functionalized magnetic microparticles: the immobilized enzyme retains its activity, greatly improves its thermostability (4 h at 75 °C), and can be recycled up to 8 times with a set of aromatic ethylamines. After the last reaction cycle, the overall conversion is about 90% for all tested substrates, with an aldehyde production ranging between 100 and 270 mg depending on the substrate used. As a proof concept, one of the aldehydes thus produced was successfully used for the biomimetic synthesis of a non-natural benzylisoquinoline alkaloid.     

Designing a personal informatics system for users without experience in self-tracking: a case study

Thanks to the advancements in ubiquitous and wearable technologies, Personal Informatics (PI) systems can now reach a larger audience of users. However, it is not still clear whether this kind of tool can fit the needs of their daily lives. Our research aims at identifying specific barriers that may prevent the widespread adoption of PI and finding solutions to overcome them. We requested users without competence in self-tracking to use different PI instruments during their daily practices, identifying five user requirements by which to design novel PI tools. On such requirements, we developed a new system that can stimulate the use of these technologies, by enhancing the perceived benefits of collecting personal data. Then, we explored how naïve and experienced users differently explore their personal data in our system through a user trial. Results showed that the system was successful at helping individuals manage and interpret their own data, validated the usefulness of the requirements found and inspired three further design opportunities that could orient the design of future PI systems.     

Influence of ethanol/water ratio in ultrasound and high‐pressure/high‐temperature phenolic compound extraction from agri‐food waste

The valorisation and management of agri‐food waste are currently hot investigation topics which probe the recovery of valuable compounds, such as polyphenols. In this study, high‐pressure/high‐temperature extraction (HPTE) and ultrasound‐assisted extraction (UAE) have been used to study the recovery of phenolic compounds from grape marc and olive pomace in hydroalcoholic solutions. The main phenolic compounds in both extracts were identified by HPLC‐DAD. Besides extraction yield (total polyphenol and flavonoid content) and the antiradical power, polyphenol degradation under HPTE and UAE has also been studied. HPTE with ethanol 75% gave higher phenolic extraction yields: 73.8 ± 1.4 mg of gallic acid equivalents per gram of dried matter and 60.0 mg of caffeic acid equivalents per gram of dried matter for grape marc and olive pomace, respectively. In this study, the efficient combination of ethanol/water mixture with HPTE or UAE has been used to enhance the recovery of phenolic compounds from grape marc and olive pomace. HPLC‐DAD showed that UAE prevents phenolic species degradation damage because of its milder operative conditions.     

Energy performance of an exhibition hall in a life cycle perspective: embodied energy,operational energy and retrofit strategies

Nowadays, the focus on the building energy consumption in the use phase prevails over an interest concerning the energy impacts linked to all the other phases of the construction process. However, reducing operational energy could lead to shifting the impacts from one stage to another. Thus, combining the study of strategies improving energy efficiency in the use phase with a life cycle approach is crucial. Exhibition halls are peculiar buildings from the geometry, construction and use points of view, rarely addressed in energy and life cycle energy analysis studies. Therefore, in this paper, a representative hall of the Milan Trade Fair is taken as a case study. A building energy simulation model is firstly calibrated in order to derive the operational energy for climatisation. The operational energy appears artificially low due to the short use period during the year. When compared with the calculated embodied energy of the envelope and structure, it is found that 57 years would be needed to balance energy spent in the construction and in the use phase. Further, some retrofit interventions are proposed and analyzed. Insulation interventions are not attractive from the economic payback time point of view. However, when the embodied energy of the retrofit interventions is compared with the energy savings in the use phase, interesting energy payback times are obtained. Therefore, this study puts in evidence on the importance of adopting a life cycle perspective, especially for buildings with low-intensity use. Eventually, the critical issues of the life cycle energy analysis are deeply discussed.     

Commiphora myrrha (Nees) Engl. extracts: evaluation of antioxidant and antiproliferative activity and their ability to reduce microbial growth on fresh‐cut salad

With the aim to develop natural preservatives displaying also chemopreventive activity, different Commiphora myrrha (Nees) Engl. extracts were studied. Myrrh essential oils, obtained by steam distillation and microwave‐assisted hydrodistillation, and several other extracts, obtained by sequential procedures with petroleum ether (PE), ethanol, ethyl acetate and butanol, have been screened for their antioxidant (DPPH· scavenging assay) and antiproliferative activity (on both nontumour and colon cancer cell lines) without previous purification. Considering that the colon cancer cell lines were more sensitive to PE and ethanol extracts, the latter of which showed the highest antioxidant activity (EC50 = 0.160 ± 0.008 mg mL^?1), both have been selected for further antibacterial/antifungal activity tests using an antimicrobial diffusion test and a growth inhibition test on salads. Results showed that the ethanol extract possessed the higher antibacterial and antifungal activity. Compared to untreated product, fresh‐cut salads treated with these two myrrh extracts displayed a significant lower bacterial growth. Although further investigation is required, these promising results offer hints as how to improve the shelf life of fresh‐cut salad.