The role of mass spectrometry in the study of non-enzymatic protein glycation in diabetes: an update

Recent studies on non-enzymatic protein glycation are reviewed, and results are critically discussed. Advanced glycation end products (AGE) levels in the body reflect a balance between their formation and catabolism. AGE proteolysis leads to the formation of low-molecular-weight AGE (AGE peptides) that are normally excreted in urine. In the case of diabetic disease and/or renal failure, AGE peptides accumulate in plasma. Because of their high reactivity, these compounds have been thought to play a role in the progression of chronic complications. The structural identification of these compounds is particularly important, and a strategy has been designed for their possible definition. A series of experiments has been devoted to the study of the enzymatic degradation products of in vitro glycated human serum albumin (HSA). This approach, based on different MS methods (LC/ESI/MS, LC/ESI/FTMS, MALDI), led to the detection of the glycated peptides generated by digestion of HSA. A further study was devoted to the possible identification of the peptides identified in the glycated HSA digestion products in the plasma of diabetic and nephropatic subjects. No glycated HSA digestion products were found in plasma samples of the subjects under investigation even if clear differences were found among the LC runs from populations of healthy, diabetic, and nephropatic subjects. Parallel investigations were devoted to the evaluation of glyoxal and methylglyoxal-dicarbonyl compounds that originate at the intermediate stage of the Maillard reaction. This evaluation was performed in diabetic patients, before and after the achievement of good metabolic control, and in nephropatic patients subjected to peritoneal dialysis (PD). In the latter case, results indicated that these dicarbonyl compounds, already present in the dialysis fluids, show a decrease in plasma and in dialysis fluids; those data suggested their reaction at peritoneal membrane level.     

Wetting behavior of graphite and CFC composites by Cu-Ti compacts

The wetting behavior of Cu-Ti powder compacts with 22 wt %Ti and 50 wt %Ti on carbon materials, including graphite and carbon fiber reinforced carbon composites(CFC), has been investigated in a vacuum using the sessile drop method. The equilibrium contact angles of Cu-22Ti(containing 22 wt%Ti) on the graphite and the CFC substrates at 1 253 K are 32 o and 26 o, respectively, whereas the equilibrium contact angle of 9° is obtained for Cu-50Ti(containing 50 wt%Ti) on both the graphite and the CFC substrates at 1 303 K.Microstructural analysis of the wetting samples shows that a thin TiC reaction layer is developed at the interfacial area and Ti-Cu intermetallic compounds are formed over the reaction layer. The investigation on the spreading kinetics of Cu-Ti compacts on carbon materials substrates at fixed temperatures reveals that the spreading is controlled by the interfacial reactions in the first stage and then by the diffusion of the active Ti from the drop bulk to the triple line in the later stage. The spreading is promoted by the intense reaction at higher Ti concentrations.     

Polymer‐derived ceramic aerogels as sorbent materials for the removal of organic dyes from aqueous solutions

Polymer‐derived SiC and SiOC aerogels have been synthesized and characterized both from the microstructural point of view and as sorbent materials for removing organic dyes (Methylene Blue, MB, and Rhodamine B, RB) from water solutions. Their adsorbent behavior has been compared with a polymer‐derived SiC foam and a commercial mesoporous silica. The aerogels can efficiently remove MB and RB from water solution and their capacity is higher compared to the SiC foams due to the higher surface area. The SiOC aerogel remains monolithic after the water treatment (allowing for an easy removal without the need of a filtration step) and its maximum capacity for removing MB is 42.2 mg/g, which is higher compared to the studied mesoporous silica and many C‐based porous adsorbents reported in the literature. The reason for this high adsorption capacity has been related to the unique structure of the polymer‐derived SiOC, which consists of an amorphous silicon oxycarbide network and a free carbon phase.     

The role played by the alpha-helix in the unfolding pathway and stability of azurin: switching between hierarchic and nonhierarchic folding

The role played by the alpha-helix in determining the structure, the stability and the unfolding mechanism of azurin was addressed by studying a helix-depleted azurin variant produced by site-directed mutagenesis. The protein structure was investigated by CD, 1D (1)H NMR, fluorescence spectroscopy measurements and MD simulations, whilst EPR, UV-visible and cyclic voltammetry experiments were carried out to investigate the geometry and the properties of the Cu(II) site. The effects of the alpha-helix depletion on the thermal stability and the unfolding pathway of the protein were determined by DSC, UV/visible and fluorescence measurements at increasing temperature. The results show that, in the absence of the alpha-helix segment, the overall protein structure is maintained, and that only the Cu site is slightly modified. In contrast, the protein stability is diminished by about 60% with respect to the wild-type azurin. Moreover, the unfolding pathway of the mutant azurin involves the presence of detectable intermediates. In comparison with previous studies concerning other small beta-sheet cupredoxins, the results as a whole support the hypothesis that the presence of the alpha-helix can switch the folding of azurin from a hierarchic to a nonhierarchic mechanism in which the highly conserved beta-sheet core provides a scaffold for cooperative folding of the wild-type protein.     

A collagenase-targeted multiplex PCR assay for identification of Vibrio alginolyticus, Vibrio cholerae, and Vibrio parahaemolyticus

A multiplex PCR assay using three collagenase-targeted primer pairs for the species-specific detection of Vibrio alginolyticus, Vibrio cholerae, and Vibrio parahaemolyticus was developed. The results highlight the species specificity of the three primer sets designed. Because of the increasing importance of Vibrio spp. in human foodborne diseases, molecular approaches for routine microbial screening and monitoring of clinical, environmental, and food samples also have become more important. The results of this study indicate that the gene coding for collagenase should be used as an alternative molecular target to discriminate among the three Vibrio species.     

Criticism on the hg-index

Although composition of bibliometric indicators appears to be desirable, in many cases it may be misleading. After a brief introduction on the properties of scales of measurement, the attention of this communication is focused on a recent composite indicator, the hg-index, suggested by Alonso et al. (Scientometrics 82(2):391?C400, 2010). Specifically, hg-index has three major criticalities: (1) the hg scale is the result of a composition of the h- and g-indices, which are defined both on ordinal scales, (2) the equivalence classes of hg are questionable and the substitution rate between h and g may arbitrarily change depending on the specific h and g values, (3) the apparent increase in granularity of hg, with respect to h and g, is illusory and misleading. Argument is supported by several examples.     

PAHs concentration in heat-treated milk samples

In this study the presence of residual levels of Polycyclic Aromatic Hydrocarbons (PAHs) in milk samples from Calabria was evaluated. A comparative analysis of PAHs concentrations was conducted on raw, pasteurized, UHT semi-skimmed and whole milk.Quantitative determination of PAHs was performed by HPLC using a fluorescence detector and analysis in HPLC-MS was conducted to confirm the presence of these compounds.Residual levels of PAHs were found in all milk samples analyzed, showing higher concentrations in pasteurized and UHT milk than in raw milk samples.The results obtained demonstrate that PAHs presence also in raw milk is dependent from environmental pollution but pasteurization and UHT treatments of milk can influence PAHs formation; the differences found between whole and semi-skimmed samples can be due to different fat content of milk.     

Inertial-magnetic sensors for assessing spatial cognition in infants

This letter describes a novel approach to the assessment of spatial cognition in children. In particular, we present a wireless instrumented toy embedding magneto-inertial sensors for orientation tracking, specifically developed to assess the ability to insert objects into holes. To be used in naturalistic environments (e.g., day cares), we also describe an in-field calibration procedure based on a sequence of manual rotations, not relying on accurate motions or sophisticated equipment. The final accuracy of the proposed system, after the mentioned calibration procedure, is derived by direct comparison with a gold-standard motion tracking device. In particular, both systems are subjected to a sequence of ten single-axis rotations (approximately 90°, back and forth), about three different axes. The rms of the angular error between the two measurements (gold-standard versus proposed systems) was evaluated for each trial. In particular, the average rms error is under 2°. This study indicates that a technological approach to ecological assessment of spatial cognition in infants is indeed feasible. As a consequence, prevention through screening of large number of infants is at reach.     

Lanthionine,a Novel Uremic Toxin,in the Vascular Calcification of Chronic Kidney Disease: The Role of Proinflammatory Cytokines

Vascular calcification (VC) is a risk factor for cardiovascular events and mortality in chronic kidney disease (CKD). Several components influence the occurrence of VC, among which inflammation. A novel uremic toxin, lanthionine, was shown to increase intracellular calcium in endothelial cells and may have a role in VC. A group of CKD patients was selected and divided into patients with a glomerular filtration rate (GFR) of <45 mL/min/1.73 m² and ≥45 mL/min/1.73 m². Total Calcium Score (TCS), based on the Agatston score, was assessed as circulating lanthionine and a panel of different cytokines. A hemodialysis patient group was also considered. Lanthionine was elevated in CKD patients, and levels increased significantly in hemodialysis patients with respect to the two CKD groups; in addition, lanthionine increased along with the increase in TCS, starting from one up to three. Interleukin IL-6, IL-8, and Eotaxin were significantly increased in patients with GFR < 45 mL/min/1.73 m² with respect to those with GFR ≥ 45 mL/min/1.73 m². IL-1b, IL-7, IL-8, IL-12, Eotaxin, and VEGF increased in calcified patients with respect to the non-calcified. IL-8 and Eotaxin were elevated both in the low GFR group and in the calcified group. We propose that lanthionine, but also IL-8 and Eotaxin, in particular, are a key feature of VC of CKD, with possible marker significance.