Mass spectrometry in the diagnosis of thyroid disease and in the study of thyroid hormone metabolism

The importance of thyroid hormones in the regulation of development, growth, and energy metabolism is well known. Over the last decades, mass spectrometry has been extensively used to investigate thyroid hormone metabolism and to discover and characterize new molecules involved in thyroid hormones production, such as thyrotropin-releasing hormone. In the earlier period, the quantification methods, usually based on gas chromatography–mass spectrometry, were complicated and time consuming. They were mainly focused on basic research, and were not suitable for clinical diagnostics on a routine basis. The development of the modern mass spectrometers, mainly coupled to liquid chromatography, enabled simpler sample preparation procedures, and the accurate quantification of thyroid hormones, of their precursors, and of their metabolites in biological fluids, tissues, and cells became feasible. Nowadays, molecules of physiological and pathological interest can be assayed also for diagnostic purposes on a routine basis, and mass spectrometry is slowly entering the clinical laboratory. This review takes stock of the advancements in the field of thyroid metabolism that were carried out with mass spectrometry, with special focus on the use of this technique for the quantification of molecules involved in thyroid diseases.     

NMR Profiling of Exhaled Breath Condensate Defines Different Metabolic Phenotypes of Non-Cystic Fibrosis Bronchiectasis

Nuclear-magnetic-resonance (NMR) profiling of exhaled breath condensate (EBC) provides insights into the pathophysiology of bronchiectasis by identifying specific biomarkers. We evaluated whether NMR-based metabolomics discriminates the EBC-derived metabolic phenotypes (“metabotypes”) of 41 patients with non-cystic fibrosis (nCF) bronchiectasis of various etiology [24 subjects with Primary Ciliary Dyskinesia (PCD); 17 patients with bronchiectasis not associated with PCD (nCF/nPCD)], who were compared to 17 healthy subjects (HS). NMR was used for EBC profiling, and Orthogonal Projections to Latent Structures with partial least-squares discriminant analysis (OPLS-DA) was used as a classifier. The results were validated by using the EBC from 17 PCD patients not included in the primary analysis. Different statistical models were built, which compared nCF/nPCD and HS, PCD and HS, all classes (nCF/nPCD-PCD-HS), and, finally, PCD and nCF/nPCD. In the PCD-nCF/nPCD model, four statistically significant metabolites were able to discriminate between the two groups, with only a minor reduction of the quality parameters. In particular, for nCF/nPCD, acetone/acetoin and methanol increased by 21% and 18%, respectively. In PCD patients, ethanol and lactate increased by 25% and 28%, respectively. They are all related to lung inflammation as methanol is found in the exhaled breath of lung cancer patients, acetone/acetoin produce toxic ROS that damage lung tissue in CF, and lactate is observed in acute inflammation. Interestingly, a high concentration of ethanol hampers cilia beating and can be associated with the genetic defect of PCD. Model validation with 17 PCD samples not included in the primary analysis correctly predicted all samples. Our results indicate that NMR of EBC discriminates nCF/nPCD and PCD bronchiectasis patients from HS, and patients with nCF/nPCD from those with PCD. The metabolites responsible for between-group separation identified specific metabotypes, which characterize bronchiectasis of a different etiology.     

Electrochemical behaviour of a magnesium alloy containing rare earth elements

The corrosion of a magnesium alloy containing rare earth elements (WE43 type alloy) was studied in 0.05 and 0.5 M Na₂SO₄ or 0.1 and 1 M NaCl solutions using electrochemical techniques: linear polarization resistance, potentiodynamic polarization, impedance measurements. The electrolytes favoured anodic magnesium oxidation but the presence of rare earth elements improved the tendency of magnesium to passivation. The dissolution rates in chlorides were higher than in sulphates because chlorides, in contrast to sulphates, interfered with the formation and maintenance of a protective layer of corrosion products which decreased the severity of the attack. The effects of galvanic corrosion due to cathodic intermetallic precipitates at grain boundaries were particularly evident in chloride media at long testing times.     

A Stingless Bee (Melipona seminigra) Marks Food Sources with a Pheromone from Its Claw Retractor Tendons

By depositing scent marks on flowers, bees reduce both the search time and the time spent with the handling of nonrewarding flowers. They thereby improve the efficiency of foraging. Whereas in honey bees the source of these scent marks is unknown, it is assumed to be the tarsal glands in bumble bees. According to histological studies, however, the tarsal glands lack any openings to the outside. Foragers of the stingless bee Melipona seminigra have previously been shown to deposit an attractant pheromone at sugar solution feeders, which is secreted at the tips of their tarsi. Here we show that the claw retractor tendons have specialized glandular epithelia within the femur and tibia of all legs that produce this pheromone. The secretion accumulates within the hollow tendon, which also serves as the duct to the outside, and is released from an opening at the base of the unguitractor plate. In choice experiments, M. seminigra was attracted by feeders baited with pentane extracts of the claw retractor tendons in the same way as it was attracted by feeders previously scent marked by foragers. Our results resolve the seeming contradiction between the importance of foot print secretions and the lack of openings of the tarsal glands.     

Melatonin prevents mitochondrial dysfunctions and death in differentiated skeletal muscle cells

Oxidative stress increase induces cellular damage and apoptosis activation, a mechanism believed to represent a final common pathway correlated to sarcopenia and many skeletal muscle disorders. The goal of this study is to evaluate if melatonin, a ROS scavenger molecule, is able to counteract or modulate myotube death. Here, differentiated C2C12 skeletal muscle cells have been treated with melatonin before chemicals known to induce apoptotic death and oxidative stress, and its effect has been investigated by means of morpho‐functional analyses. Ultrastructural observations show melatonin protection against triggers by the reducing of membrane blebbing, chromatin condensation, myonuclei loss and in situ DNA cleavage. Moreover, melatonin is able to prevent mitochondrial dysfunctions which occur in myotubes exposed to the trigger alone. These findings demonstrate melatonin ability in preventing apoptotic cell death in skeletal muscle fibers in vitro, suggesting for this molecule a potential therapeutic role in the treatment of various muscle disorders.     

Sulfolane biodegradation potential in aquifer sediments at sour natural gas plant sites

Sulfolane is used in the treatment of sour natural gas. It is a highly water soluble compound that has been introduced into soils and groundwaters at a number of sour gas processing plant sites. Aquifer sediments from contaminated locations at three sites in western Canada were assessed for microbial activity and their ability to degrade sulfolane under aerobic and five anaerobic (nitrate-, Mn(IV)-, Fe(III)-, sulfate- and CO₂-reducing) conditions. The microcosms were supplemented with 200 mg/L sulfolane and adequate supplies of N, P, and the appropriate terminal electron acceptor. Microcosms containing contaminated aquifer sediments from each of the three sites were able to degrade sulfolane aerobically at 8°C and 28°C, and the biodegradation followed zero-order kinetics. The lag times before the onset of sulfolane biodegradation were shorter when sulfolane-contaminated sediments were used as inocula than when uncontaminated soils were used. No anaerobic sulfolane biodegradation was observed at 28°C, nor was sulfolane biodegradation observed at 8°C under Fe(III)-, sulfate- and CO₂-reducing conditions. At 8°C, anaerobic degradation of sulfolane coupled to Mn(IV) reduction was observed in microcosms from two sites, and degradation coupled to nitrate reduction was seen in a microcosm from one of the contaminated sites.     

Stress corrosion cracking of 13% Cr martensitic steels in sodium chloride solutions in the presence of thiosulphate

The stress corrosion cracking (SCC) susceptibility of 13% Cr martensitic (UNS S42000) and supermartensitic (UNS S41125) steels in sodium chloride solutions in the presence of thiosulphate was evaluated by slow strain rate tests (SSRT). The tests were performed in 5% sodium chloride solutions buffered at pH 2.7, 3.5, 4.5 and 6.0 in the absence and presence of thiosulphate in a concentration range between 10⁻⁶ and 10⁻³ M, at 25 ± 0.1 °C. The electrochemical behaviour of the two steels in the different solutions was determined by recording the anodic and cathodic polarisation curves. 13% Cr martensitic steel showed SCC in 5% sodium chloride solutions with pH ≤ 4.5 in the presence of 3 × 10⁻⁶ M thiosulphate. Decreasing the chloride ion concentration from 50 to 10 g/l, the critical concentration of thiosulphate to provoke SCC susceptibility increased from 3 × 10⁻⁶ to 1 × 10⁻⁵ M. The resistance to SCC of the supermartensitic steel was higher than that of the martensitic steel. The critical concentration of thiosulphate to induce SCC on the supermartensitic steel were 1 × 10⁻⁵ M at pH 2.7 and 1 × 10⁻⁴ M at pH 3.5. At pH ≥ 4.5 the supermartensitic steel did not crack. The anodic and cathodic polarisation curves evidenced the influence of the thiosulphate on the corrosion and the activation effect on the steels. The SCC of the two steels was attributed to hydrogen embrittlement produced by sulphur and hydrogen sulphide formed by dismutation and reduction of thiosulphate.     

Lipid Cubic Mesophases Combined with Superparamagnetic Iron Oxide Nanoparticles: A Hybrid Multifunctional Platform with Tunable Magnetic Properties for Nanomedical Applications

Hybrid materials composed of superparamagnetic iron oxide nanoparticles (SPIONs) and lipid self-assemblies possess considerable applicative potential in the biomedical field, specifically, for drug/nutrient delivery. Recently, we showed that SPIONs-doped lipid cubic liquid crystals undergo a cubic-to-hexagonal phase transition under the action of temperature or of an alternating magnetic field (AMF). This transition triggers the release of drugs embedded in the lipid scaffold or in the water channels. In this contribution, we address this phenomenon in depth, to fully elucidate the structural details and optimize the design of hybrid multifunctional carriers for drug delivery. Combining small-angle X-ray scattering (SAXS) with a magnetic characterization, we find that, in bulk lipid cubic phases, the cubic-to-hexagonal transition determines the magnetic response of SPIONs. We then extend the investigation from bulk liquid-crystalline phases to colloidal dispersions, i.e., to lipid/SPIONs nanoparticles with cubic internal structure (“magnetocubosomes”). Through Synchrotron SAXS, we monitor the structural response of magnetocubosomes while exposed to an AMF: the magnetic energy, converted into heat by SPIONs, activates the cubic-to-hexagonal transition, and can thus be used as a remote stimulus to spike drug release “on-demand”. In addition, we show that the AMF-induced phase transition in magnetocubosomes steers the realignment of SPIONs into linear string assemblies and connect this effect with the change in their magnetic properties, observed at the bulk level. Finally, we assess the internalization ability and cytotoxicity of magnetocubosomes in vitro on HT29 adenocarcinoma cancer cells, in order to test the applicability of these smart carriers in drug delivery applications.