To shift, or not to shift: Adequate selection of an internal standard in mass-shift approaches using tandem ICP-mass spectrometry (ICP-MS/MS)

The use of an internal standard to correct for potential matrix effects and instrument instability is common practice in ICP-MS. However, with the introduction of a new generation of ICP-MS instrumentation with a tandem mass spectrometry configuration (ICP-MS/MS), the use of chemical resolution in a mass-shift approach has become much more popular, suggesting that the appropriate selection of an internal standard needs revision. In this particular case, it needs to be decided whether the internal standard should also be subjected to a mass-shift or can simply be monitored on-mass ("to shift, or not to shift"). In this work, 17 elements covering a wide range of masses (24-205 amu) and ionization energies (3.89-9.39 eV) were measured via on-mass and/or mass-shift strategies, and the corresponding atomic ions and reaction product ions were monitored during various systematic experiments. For mass-shifting, an NH3/He gas mixture was used to obtain NH3-based reaction product ions (cluster formation). Product ion scanning (PIS) was used for assessing the differences in reactivity between the different analytes and for the identification of the best suited reaction product ions. It was found that the use of chemical resolution can significantly affect the short-term signal stability and that ion signals measured on-mass are not affected in the same way as those measured mass-shifted. Variations affecting the signal intensities of both atomic and reaction product ions can be attributed to the ion-molecule chemistry occurring within the collision/reaction cell and were found to be related with some degree of initial instability in the cell and differences in reactivity. The use of a sufficiently long stabilization time, however, avoids or at least mitigates such differences in the behavior between signals monitored on-mass and after mass-shifting, respectively. Furthermore, the introduction of cell disturbances, such as those generated after quickly switching between different sets of operating conditions in a multi-tune method, revealed significant differences in signal behavior between atomic and reaction product ions, potentially hampering the use of an internal standard monitored on-mass when the analysis is based on an analyte monitored after mass-shifting. However, the use of a reasonable waiting time again greatly mitigates such differences, with the duration of this stabilization time depending on the magnitude of the cell disturbances (e.g., switch between vented and pressurized mode or only between pressurized modes using different gas flow rates). In addition, also the effect of varying different instrument settings (plasma power, torch position, and gas and liquid flow rates) was evaluated, but no remarkable differences were found between signals monitored on-mass and those mass-shifted. Interestingly, a statistical evaluation of the influence of the different settings on the signal intensities of all ions monitored did not reveal the a priori important role of some properties traditionally suggested for adequate selection of analyte/internal standard pairs, such as mass number or ionization energy, as also suggested in other recent studies. © The Royal Society of Chemistry

Parallel Readout of Pathway-Specific Inputs to Laminated Brain Structures

Local field potentials (LFPs) capture the electrical activity produced by principal cells during integration of converging synaptic inputs from multiple neuronal populations. However, since synaptic currents mix in the extracellular volume, LFPs have complex spatiotemporal structure, making them hard to exploit. Here we propose a biophysical framework to identify and separate LFP-generators. First we use a computational multineuronal model that scales up single cell electrogenesis driven by several synaptic inputs to realistic aggregate LFPs. This approach relies on the fixed but distinct locations of synaptic inputs from different presynaptic populations targeting a laminated brain structure. Thus the LFPs are contributed by several pathway-specific LFP-generators, whose electrical activity is defined by the spatial distribution of synaptic terminals and the time course of synaptic currents initiated in target cells by the corresponding presynaptic population. Then we explore the efficacy of independent component analysis to blindly separate converging sources and reconstruct pathway-specific LFP-generators. This approach can optimally locate synaptic inputs with subcellular accuracy while the reconstructed time course of pathway-specific LFP-generators is reliable in the millisecond scale. We also describe few cases where the non-linear intracellular interaction of strongly overlapping LFP-generators may lead to a significant cross-contamination and the appearance of derivative generators. We show that the approach reliably disentangle ongoing LFPs in the hippocampus into contribution of several LFP-generators. We were able to readout in parallel the pathway-specific presynaptic activity of projection cells in the entorhinal cortex and pyramidal cells in the ipsilateral and contralateral CA3. Thus we provide formal mathematical and experimental support for parallel readout of the activity of converging presynaptic populations in working neuronal circuits from common LFPs

Evaluating the repetitive mucus extraction effects on mucus biomarkers, mucous cells, and the skin-barrier status in a marine fish model

Among all the mucosal barriers, the skin and its surrounding mucus are possibly the main defensive tool against changes in the environment that can be harmful for fish. Due to the extraction of this mucus being less invasive, the study of its production and functions has attracted great interest in recent years. However, there are still many gaps concerning the sampling process as well as the possible alterations in skin integrity and mucus composition. In the current study, the effects of skin mucus extraction were determined by comparing the effects of a single extraction (single extraction group, SEG) with those of three successive extractions separated by 3 days (repetitive extractions group, REG). Intact skin histology without mucus extraction (ØEG) and both plasma and skin mucus biomarkers and antibacterial capacities were also assessed. Regarding the skin histology and skin barrier properties, both the SEG and REG did not show differences in the intact skin. Interestingly, repetitive mucus extractions seemed to activate skin mucus turnover, significantly increasing the number of small-sized mucous cells (cell area 150 µm2). Repetitive extractions significantly decreased the amounts of soluble protein and increased cortisol secretion. These metabolites remained unaltered in the plasma, indicating different responses in the plasma and mucus. Despite changes in the mucus biomarkers, antibacterial capacity against pathogenic bacteria (Pseudomonas anguilliseptica and Vibrio anguillarum) was maintained in both the plasma and mucus irrespective of the number of mucus extractions. Overall, the mucus sampling protocol had little effect on skin integrity and mucus antibacterial properties, only modifying the amounts of soluble protein exuded and stimulating mucous cell replacement. This protocol is a feasible and minimally invasive way of studying and monitoring fish health and welfare and can be used as an alternative or a complement to plasma analysis. This methodology can be transferred to farm culture conditions and be very useful for studying threatened species in order to preserve fish welfare.info:eu-repo/semantics/publishedVersio