Comparison of the fluctuations of the signals measured by ICP-MS after laser ablation of powdered geological materials prepared by four methods.

Sample preparation is a crucial point for quantitative multi-elemental analses by LA-ICP-MS of powdered geological materials. Four different methods are compared in this study with respect to signal stability and intensity as follows: the preparation of glass beads (GlassB) by alkaline fusion method and three grinding and pelletizing methods relying on the use of an organic binder (VanBind, vanillic acid), an adhesive binder (MixGlue, methyl methacrylate) and a sol-gel process for glass formation (SolGel, chemical reaction of tetraethoxysilane), respectively. Sixty elements were analyzed by means of a ns-UV (213 nm) laser ablation system coupled to a single collector sector field ICP-MS with a low or medium mass resolution. Signal stability was found to strongly depend on the sample homogeneity provided by the preparation method. These methods were applied to three geological standard materials (CRM). The following criteria were used to evaluate and compare the methods: (1) proportion of the measurement cycles which are above a given signal intensity threshold (defined here as signal average ± 3 times the standard deviation), (2) signal stability of the analyzed nuclides (internal precision estimated by the relative standard deviations on raw count rates), (3) signal stability of the internal standards added to the samples, (4) external precision estimated by the relative standard deviation over five preparations for each geological CRM. For the majority of the analyzed elements, signals measured for samples prepared with the four methods are reproducible. Specific contamination in one or several elements (Cr, Fe, Co, Ni, Cu, Mo, W, Au and Bi) was observed depending on the sample preparation method. In addition, compared to grinding made with PTFE material, grinding performed with tungsten carbide material was found to produce better homogeneity, especially for the sol-gel and mixing with glue protocols, although some metallic contamination (W and Co) was observed. Thanks to the suppression of grain effects by alkaline melting, the glass bead method systematically provided signal stability and percentage of "over the threshold" close to those of the NIST glasses. This may be explained by the preparation of more homogeneous samples by alkaline melting. Finally, the described methods were found to be reproducible for the majority of the analyzed elements.

From a Cone Snail Toxin to a Competitive MC4R Antagonist.

The melanocortin 4 receptor (MC4R) plays a role in energy homeostasis and represents a target for treating energy balance disorders. For decades, synthetic ligands have been derived from MC4R endogenous agonists and antagonists, such as setmelanotide used to treat rare forms of genetic obesity. Recently, animal venoms have demonstrated their capacity to provide melanocortin ligands with toxins from a scorpion and a spider. Here, we described a cone snail toxin, N-CTX-Ltg1a, with a nanomolar affinity for hMC4R but unrelated to any known toxins or melanocortin ligands. We then derived from the conotoxin the linear peptide HT1-0, a competitive antagonist of Gs, G15, and ß-arrestin2 pathways with a low nanomolar affinity for hMC4R. Similar to endogenous ligands, HT1-0 needs hydrophobic and basic residues to bind hMC4R. Altogether, it represents the first venom-derived peptide of high affinity on MC4R and paves the way for the development of new MC4R antagonists.

A snake toxin as a theranostic agent for the type 2 vasopressin receptor.

Rationale: MQ1, a snake toxin which targets with high nanomolar affinity and absolute selectivity for the type 2 vasopressin receptor (V2R), is a drug candidate for renal diseases and a molecular probe for imaging cells or organs expressing V2R. Methods: MQ1's pharmacological properties were characterized and applied to a rat model of hyponatremia. Its PK/PD parameters were determined as well as its therapeutic index. Fluorescently and radioactively labeled MQ1 were chemically synthesized and associated with moderate loss of affinity. MQ1's dynamic biodistribution was monitored by positron emission tomography. Confocal imaging was used to observe the labeling of three cancer cell lines. Results: The inverse agonist property of MQ1 very efficiently prevented dDAVP-induced hyponatremia in rats with low nanomolar/kg doses and with a very large therapeutic index. PK (plasma MQ1 concentrations) and PD (diuresis) exhibited a parallel biphasic decrease. The dynamic biodistribution showed that MQ1 targets the kidneys and then exhibits a blood and kidney biphasic decrease. Whatever the approach used, we found a T1/2α between 0.9 and 3.8 h and a T1/2ß between 25 and 46 h and demonstrated that the kidneys were able to retain MQ1. Finally, the presence of functional V2R expressed at the membrane of cancer cells was, for the first time, demonstrated with a specific fluorescent ligand. Conclusion: As the most selective V2 binder, MQ1 is a new promising drug for aquaresis-related diseases and a molecular probe to visualize in vitro and in vivo V2R expressed physiologically or under pathological conditions.