VDACs Post-Translational Modifications Discovery by Mass Spectrometry: Impact on Their Hub Function

VDAC (voltage-dependent anion selective channel) proteins, also known as mitochondrial porins, are the most abundant proteins of the outer mitochondrial membrane (OMM), where they play a vital role in various cellular processes, in the regulation of metabolism, and in survival pathways. There is increasing consensus about their function as a cellular hub, connecting bioenergetics functions to the rest of the cell. The structural characterization of VDACs presents challenging issues due to their very high hydrophobicity, low solubility, the difficulty to separate them from other mitochondrial proteins of similar hydrophobicity and the practical impossibility to isolate each single isoform. Consequently, it is necessary to analyze them as components of a relatively complex mixture. Due to the experimental difficulties in their structural characterization, post-translational modifications (PTMs) of VDAC proteins represent a little explored field. Only in recent years, the increasing number of tools aimed at identifying and quantifying PTMs has allowed to increase our knowledge in this field and in the mechanisms that regulate functions and interactions of mitochondrial porins. In particular, the development of nano-reversed phase ultra-high performance liquid chromatography (nanoRP-UHPLC) and ultra-sensitive high-resolution mass spectrometry (HRMS) methods has played a key role in this field. The findings obtained on VDAC PTMs using such methodologies, which permitted an in-depth characterization of these very hydrophobic trans-membrane pore proteins, are summarized in this review.     

Sardinian honeys as sources of xanthine oxidase and tyrosinase inhibitors

Sardinian honeys obtained from different floral sources (Arbutus, Asphodelus, Eucalyptus, Thistle, and Sulla) were evaluated for their ability to inhibit tyrosinase and xanthine oxidase enzymes and for their antioxidant activity. Physicochemical parameters, total phenolic, and flavonoids content were also determined. Honey from Arbutus flowers had the highest antioxidant activity followed by Eucalyptus and Thistle ones. These three honeys showed good tyrosinase and xanthine oxidase inhibition properties. Thus, these Sardinian honeys could have a great potential as antioxidant sources for pharmaceutical and cosmetic applications.     

Nonthermal Technologies for Fruit and Vegetable Juices and Beverages: Overview and Advances

In recent years, there has been a growing interest in the design of novel nonthermal processing systems that minimally modify sensory, nutritional, and functional properties of fruit and vegetable juices and beverages. The benefits of nonthermal treatments are strongly dependent on the food matrix. Thus, an understanding of the effects that these technologies exert on the properties of juices and beverages is important to design and optimize technological parameters to produce value‐added products. This review covers research on nonthermal electrical treatments, high pressure processing, ultrasound, radiation processing, inert gas treatments, cold plasma, and membrane processing. Advances towards optimization of processing conditions, and combined technologies approaches have been also extensively reviewed. This information could be useful to: (1) manage processing systems and optimize resources; (2) preserve nutritional value and organoleptic properties, and (3) provide processing conditions for validation of these technologies at the industrial scale.     

The reaction of hydrogen atoms with methionine residues: A model of reductive radical stress causing tandem protein-lipid damage

The occurrence of tandem damage, due to reductive radical stress involving proteins and lipids, is shown by using a biomimetic model. It is made of unsaturated lipid vesicle suspensions in phosphate buffer in the presence of methionine, either as a single amino acid or as part of a protein such as RNase A, which contains four methionine residues. The radical process starts with the formation of H(.) atoms by reaction of solvated electrons with dihydrogen phosphate anions, which selectively attack the thioether function of methionine. The modification of methionine to alpha-aminobutyric acid is accompanied by the formation of thiyl radicals, which in turn cause the isomerization of the cis fatty acid residues to the trans isomers. The relationship between methionine modification and lipid damage and some details of the reductive radical stress obtained by proteomic analysis of irradiated RNase A are presented.     

BIPV: merging the photovoltaic with the construction industry

BIPV (Building Integrated Photovoltaics) is a multifunctional technology that unifies the photovoltaic module with the overall building outer surface providing the building with several new functions while producing a portion or total building electricity usage. Increasing the aesthetic, functional and environmental value of a building at a much lower cost than in the recent past, new PV technologies will soon originate a market growth as intense as the growth of traditional PV market has been in the last 5 years. Copyright © 2009 John Wiley & Sons, Ltd.

     

Tokamak D-T neutron source models for different plasma physics confinement modes

Neutronic studies of European demonstration fusion power plant (DEMO) have been so far based on plasma physics low confinement mode (L-mode). Future tokamaks, nevertheless, may likely use alternative confinement modes such as high or advanced confinement modes (H&A-mode). Based on analytical formulae used in plasma physics, H&A-modes D-T neutron sources formulae are proposed in this paper. For that purpose, a tokamak random neutron source generator, TRANSGEN, has been built generating bidimensional (radial and poloidal) neutron source maps to be used as input for neutronics Monte-Carlo codes (TRIPOLI-4 and MCNP5). The impact of such a source on the neutronic behavior of the European DEMO-2007 Helium-cooled lithium–lead reactor concept has been assessed and compared with previous results obtained using a L-mode neutron source. An A-mode neutron source map from TRANSGEN has been used with the code TRIPOLI-4. Assuming the same fusion power, results show that main reactor global neutronic parameters, e.g. tritium breeding ratio and neutron multiplication factor, evolved slightly when compared to present uncertainties margin. However, local parameters, such as the neutron wall loading (NWL), change significantly compared to L-mode shape: from ?22% to +11% for NWL.     

Benchmarking of the 3-D CAD-based Discrete Ordinates code “ATTILA” for dose rate calculations against experiments and Monte Carlo calculations

Shutdown dose rate (SDDR) inside and around the diagnostics ports of ITER is performed at PPPL/UCLA using the 3-D, FEM, Discrete Ordinates code, ATTILA, along with its updated FORNAX transmutation/decay gamma library. Other ITER partners assess SDDR using codes based on the Monte Carlo (MC) approach (e.g. MCNP code) for transport calculation and the radioactivity inventory code FISPACT or other equivalent decay data libraries for dose rate assessment. To reveal the range of discrepancies in the results obtained by various analysts, an extensive experimental and calculation benchmarking effort has been undertaken to validate the capability of ATTILA for dose rate assessment. On the experimental validation front, the comparison was performed using the measured data from two SDDR experiments performed at the FNG facility, Italy. Comparison was made to the experimental data and to MC results obtained by other analysts. On the calculation validation front, the ATTILA's predictions were compared to other results at key locations inside a calculation benchmark whose configuration duplicates an upper diagnostics port plug (UPP) in ITER. Both serial and parallel version of ATTILA-7.1.0 are used in the PPPL/UCLA analysis performed with FENDL-2.1/FORNAX databases. In the FNG 1st experimental, it was shown that ATTILA's dose rates are largely over estimated (by ～30–60%) with the ANSI/ANS-6.1.1 flux-to-dose factors whereas the ICRP-74 factors give better agreement (10–20%) with the experimental data and with the MC results at all cooling times. In the 2nd experiment, there is an under estimation in SDDR calculated by both MCNP and ATTILA based on ANSI/ANS-6.1.1 for cooling times up to ～4 days after irradiation. Thereafter, an over estimation is observed (～5–10% with MCNP and ～10–15% with ATTILA). As for the calculation benchmark, the agreement is much better based on ICRP-74 1996 data. The divergence among all dose rate results at ～11 days cooling time is no more than 15% among all participants.     

Heat Transfer in Water-Based SiC and TiO2 Nanofluids

The article reports the results of heat transfer experimental tests on water-based TiO₂ (9 wt%) and SiC (3, 6, 9 wt%) nanofluids. Measurements were performed in a two-loop test rig for immediate comparison of the thermal performances of the nanofluid with the base fluid. The convective heat transfer is evaluated in a circular pipe heated with uniform heat flux (from 20 to 240 kW/m²) and flow regimes from laminar to turbulent. Tests have been performed to compare the heat transfer of nanofluids and water at the same velocity (from 0.7 to 1.6 m/s) or Reynolds number (from 300 to 6000), and they have also been compared with values calculated from some of the most widely used correlations. The analysis of the experimental data shows a strong dependence on the parameter used, while both the nanofluid and water data have the same agreement with the calculated values. Nanofluids were manufactured through a two-step procedure: laser synthesis of nanoparticles followed by dispersion in water.     

Modulatory Effects of Polyphenols on Apoptosis Induction: Relevance for Cancer Prevention

Polyphenols, occurring in fruit and vegetables, wine, tea, extra virgin olive oil, chocolate and other cocoa products, have been demonstrated to have clear antioxidant properties in vitro, and many of their biological actions have been attributed to their intrinsic reducing capabilities. However, it has become clear that, in complex biological systems, polyphenols exhibit several additional properties which are yet poorly understood. Apoptosis is a genetically controlled and evolutionarily conserved form of cell death of critical importance for the normal embryonic development and for the maintenance of tissue homeostasis in the adult organism. The malfunction of the death machinery may play a primary role in various pathological processes, since too little or too much apoptosis can lead to proliferative or degenerative diseases, respectively. Cancer cells are characterized by a deregulated proliferation, and/or an inability to undergo programmed cell death. A large body of evidence indicates that polyphenols can exert chemopreventive effects towards different organ specific cancers, affecting the overall process of carcinogenesis by several mechanisms: inhibition of DNA synthesis, modulation of ROS production, regulation of cell cycle arrest, modulation of survival/proliferation pathways. In addition, polyphenols can directly influence different points of the apoptotic process, and/or the expression of regulatory proteins. Although the bulk of data has been obtained in in vitro systems, a number of clinical studies suggesting a preventive and therapeutic effectiveness of polyphenols in vivo is available. However, a deeper knowledge of the underlying mechanisms responsible for the modulation of apoptosis by polyphenols, and their real effectiveness, is necessary in order to propose them as potential chemopreventive and chemotherapeutic candidates for cancer treatment.     

Biological and technical variables affecting immunoassay recovery of cytokines from human serum and simulated vaginal fluid: a multicenter study

The increase of proinflammatory cytokines in vaginal secretions may serve as a surrogate marker of unwanted inflammatory reaction to microbicide products topically applied for the prevention of sexually transmitted diseases, including HIV-1. Interleukin (IL)-1beta and IL-6 have been proposed as indicators of inflammation and increased risk of HIV-1 transmission; however, the lack of information regarding detection platforms optimal for vaginal fluids and interlaboratory variation limit their use for microbicide evaluation and other clinical applications. This study examines fluid matrix variants relevant to vaginal sampling techniques and proposes a model for interlaboratory comparisons across current cytokine detection technologies. IL-1beta and IL-6 standards were measured by 12 laboratories in four countries, using 14 immunoassays and four detection platforms based on absorbance, chemiluminescence, electrochemiluminescence, and fluorescence. International reference preparations of cytokines with defined biological activity were spiked into (1) a defined medium simulating the composition of human vaginal fluid at pH 4.5 and 7.2, (2) physiologic salt solutions (phosphate-buffered saline and saline) commonly used for vaginal lavage sampling in clinical studies of cytokines, and (3) human blood serum. Assays were assessed for reproducibility, linearity, accuracy, and significantly detectable fold difference in cytokine level. Factors with significant impact on cytokine recovery were determined by Kruskal-Wallis analysis of variance with Dunn's multiple comparison test and multiple regression models. All assays showed acceptable intra-assay reproducibility; however, most were associated with significant interlaboratory variation. The smallest reliably detectable cytokine differences ( P < 0.05) derived from pooled interlaboratory data varied from 1.5- to 26-fold depending on assay, cytokine, and matrix type. IL-6 but not IL-1beta determinations were lower in both saline and phosphate-buffered saline as compared to vaginal fluid matrix, with no significant effect of pH. The (electro)chemiluminescence-based assays were most discriminative and consistently detected <2-fold differences within each matrix type. The Luminex-based assays were less discriminative with lower reproducibility between laboratories. These results suggest the need for uniform vaginal sampling techniques and a better understanding of immunoassay platform differences and cross-validation before the biological significance of cytokine variations can be validated in clinical trials. This investigation provides the first standardized analytic approach for assessing differences in mucosal cytokine levels and may improve strategies for monitoring immune responses at the vaginal mucosal interface.