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.     

A cross‐cultural comparison of colour emotion for two‐colour combinations

Psychophysical experiments were conducted in the UK, Taiwan, France, Germany, Spain, Sweden, Argentina, and Iran to assess colour emotion for two‐colour combinations using semantic scales warm/cool, heavy/light, active/passive, and like/dislike. A total of 223 observers participated, each presented with 190 colour pairs as the stimuli, shown individually on a cathode ray tube display. The results show consistent responses across cultures only for warm/cool, heavy/light, and active/passive. The like/dislike scale, however, showed some differences between the observer groups, in particular between the Argentinian responses and those obtained from the other observers. Factor analysis reveals that the Argentinian observers preferred passive colour pairs to active ones more than the other observers. In addition to the cultural difference in like/dislike, the experimental results show some effects of gender, professional background (design vs. nondesign), and age. Female observers were found to prefer colour pairs with high‐lightness or low‐chroma values more than their male counterparts. Observers with a design background liked low‐chroma colour pairs or those containing colours of similar hue more than nondesign observers. Older observers liked colour pairs with high‐lightness or high‐chroma values more than young observers did. Based on the findings, a two‐level theory of colour emotion is proposed, in which warm/cool, heavy/light, and active/passive are identified as the reactive‐level responses and like/dislike the reflective‐level response. © 2010 Wiley Periodicals, Inc. Col Res Appl, 2012     

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.     

Intrinsic Fluorescence of the Active and the Inactive Functional Forms of Human Thymidylate Synthase

The observables associated with protein intrinsic fluorescence – spectra, time decays, anisotropies – offer opportunities to monitor in real time and non-invasively a protein‘s functional form and its interchange with other forms with different functions. We employed these observables to sketch the fluorometric profiles of two functional forms of human thymidylate synthase (hTS), a homodimeric enzyme crucial for cell proliferation and thus targeted by anticancer drugs. The protein takes an active and an inactive form. Stabilization of the latter by peptides that, unlike classical hTS inhibitors, bind it at the monomer/monomer interface offers an alternative inhibition mechanism that promises to avoid the onset of drug resistance in anticancer therapy. The fluorescence features depicted herein can be used as tools to identify and quantify each of the two protein forms in solution, thus making it possible to investigate the kinetic and thermodynamic aspects of the active/inactive conformational interchange. Two examples of fluorometrically monitored interconversion kinetics are provided.     

Effect of muscle,ageing time and modified atmosphere packaging conditions on the colour,oxidative and microbiological stability of packed beef

The objective of this study was to evaluate the influence of two different vacuum ageing times (7 and 14 days) and the impact of the modified atmosphere packaging (MAP) configuration (gas/product ratios: 0.5 and 1 and gas composition: 70% O₂ + 30% CO₂ and 40% O₂ + 30% CO₂ + 30% N₂) on the quality of fresh beef during subsequent storage at 4 °C. For this purpose, three separate experiments were performed. For each experiment, two different muscles (Longissimus dorsi and Biceps femoris) were sampled from four double‐muscled Belgian Blue beef carcasses. Next to colour, also the evolution in microbial load, pH, O₂ and CO₂ in the headspace and lipid oxidation at the meat surface were evaluated. A vacuum ageing for 14 days compared with 7 days resulted in a higher initial microbial load on the day of MAP packaging, which resulted finally in a significantly shorter shelf life. This ageing effect was less pronounced on the colour stability and lipid oxidation of the meat samples. No significant influence of the packaging configuration on any of the analysed parameters (colour, microbial load, pH and lipid oxidation at the meat surface) was observed.     

The stability of vitamin A in fortified palm olein during extended storage and thermal treatment

The stability of vitamin A in Refined Bleached Deodorised Palm Olein (RBDPOL) was studied for 24 months. Vitamin A decreased with time, temperature and thermal treatment (frying/cooking). RBDPOL fortification was observed over several temperature ranges, using PET, nylon and HDPE commercial packaging materials. After 24 months, the following vitamin A contents of 39–43 IU g^?1 (39–45%) at 16–20 °C; 35–40 IU g^?1 (43–49%) at 24–29 °C; and 28–39 IU g^?1 (45–73%) at 24–45 °C were detected at the respective temperature ranges. Results showed stability of vitamin A fortified RBDPOL vegetable oil was not stable over typical shelf life (12 months). Depletion of vitamin A accelerated when the RBDPOL vegetable oil was subjected to high temperature thermal treatment.     

Evaluation of <Emphasis Type="Italic">flaA</Emphasis> Short Variable Region Sequencing,Multilocus Sequence Typing,and Fourier Transform Infrared Spectroscopy for Discrimination between <Emphasis Type="Italic">Campylobacter jejuni</Emphasis> Strains

Discriminatory and robust typing methods are needed to improve the understanding of the dynamics of food-borne Campylobacter infections and epidemiology in primary animal production. To evaluate the strain discriminatory potential of typing methods, flaA short variable region (SVR) sequencing and Fourier transform infrared (FTIR) spectroscopy were applied on a collection of 102 epidemiologically related and unrelated Campylobacter jejuni strains. Previous application of FTIR spectroscopy for subtyping of Campylobacter has been limited. A subset of isolates, initially discriminated by flaA SVR sequencing, were further subjected to multilocus sequence typing (MLST). It was found that flaA SVR sequencing had a slightly higher discriminatory power than FTIR spectroscopy, based on the Simpson diversity index. The clustering of strains indicated that FTIR spectroscopy is indeed a suitable method for discrimination of Campylobacter. The isolates were assigned to six clusters based on flaA SVR sequences and nine clusters based on the FTIR spectroscopy profiles. Furthermore, the cluster analysis of flaA SVR sequences, MLST, and FTIR spectroscopy profiles showed a high degree of congruence, assigning the isolates to similar cluster structures. In conclusion, FTIR spectroscopy can be applied for subtyping of Campylobacter, and the high discriminatory potential of both flaA SVR sequencing and FTIR spectroscopy render them suitable screening methods for large numbers of strains.     

Dual detection biosensor based on porous silicon substrate

Due to the high surface-to-volume ratio (hundreds of m²/cm³) porous silicon became during the last years a good candidate material as substrate for biosensor application. Moreover, the versatility of surface chemistry allows different functionalization approaches and large number of molecules to be captured on well-defined areas. This paper reports a dual detection method for protein recognition processes developed on different nanostructured porous silicon (PS) substrates, based on using two complementary spectroscopic techniques: fluorescence and electrochemical impedance. The structures were tested for biomolecular recognition – biotin–strepavidin couples – in order to achieve an optimum surface for protein's immobilizations. Comparative analyses of the attachment degree and preservation of the biomolecules activity on the porous silicon surfaces and silicon slides are also described.     

Towards Automated Binding Affinity Prediction Using an Iterative Linear Interaction Energy Approach

Binding affinity prediction of potential drugs to target and off-target proteins is an essential asset in drug development. These predictions require the calculation of binding free energies. In such calculations, it is a major challenge to properly account for both the dynamic nature of the protein and the possible variety of ligand-binding orientations, while keeping computational costs tractable. Recently, an iterative Linear Interaction Energy (LIE) approach was introduced, in which results from multiple simulations of a protein-ligand complex are combined into a single binding free energy using a Boltzmann weighting-based scheme. This method was shown to reach experimental accuracy for flexible proteins while retaining the computational efficiency of the general LIE approach. Here, we show that the iterative LIE approach can be used to predict binding affinities in an automated way. A workflow was designed using preselected protein conformations, automated ligand docking and clustering, and a (semi-)automated molecular dynamics simulation setup. We show that using this workflow, binding affinities of aryloxypropanolamines to the malleable Cytochrome P450 2D6 enzyme can be predicted without a priori knowledge of dominant protein-ligand conformations. In addition, we provide an outlook for an approach to assess the quality of the LIE predictions, based on simulation outcomes only.