Evaluation of the multi-element capabilities of collision/reaction cell inductively coupled plasma - mass spectrometry in wine analysis

This work explores the multi-element capabilities of inductively coupled plasma - mass spectrometry with collision/reaction cell technology (CCT-ICP-MS) for the simultaneous determination of both spectrally interfered and non-interfered nuclides in wine samples using a single set of experimental conditions. The influence of the cell gas type (i.e. He, He+H2 and He+NH3), cell gas flow rate and sample pre-treatment (i.e. water dilution or acid digestion) on the background-equivalent concentration (BEC) of several nuclides covering the mass range from 7 to 238 u has been studied. Results obtained in this work show that, operating the collision/reaction cell with a compromise cell gas flow rate (i.e. 4 mL min−1) improves BEC values for interfered nuclides without a significant effect on the BECs for non-interfered nuclides, with the exception of the light elements Li and Be. Among the different cell gas mixtures tested, the use of He or He+H2 is preferred over He+NH3 because NH3 generates new spectral interferences. No significant influence of the sample pre-treatment methodology (i.e. dilution or digestion) on the multi-element capabilities of CCT-ICP-MS in the context of simultaneous analysis of interfered and non-interfered nuclides was observed. Nonetheless, sample dilution should be kept at minimum to ensure that light nuclides (e.g. Li and Be) could be quantified in wine. Finally, a direct 5-fold aqueous dilution is recommended for the simultaneous trace and ultra-trace determination of spectrally interfered and non-interfered elements in wine by means of CCT-ICP-MS. The use of the CCT is mandatory for interference-free ultra-trace determination of Ti and Cr. Only Be could not be determined when using the CCT due to a deteriorated limit of detection when compared to conventional ICP-MS.

A third cluster of red supergiants in the vicinity of the massive cluster RSGC3

Context. Recent studies have shown that the area around the massive, obscured cluster RSGC3 may harbour several clusters of red supergiants. Aims. We analyse a clump of photometrically selected red supergiant candidates 20′ south of RSGC3 in order to confirm the existence of another of these clusters. Methods. Using medium-resolution infrared spectroscopy around 2.27 μm, we derived spectral types and velocities along the line of sight for the selected candidates, confirming their nature and possible association. Results. We find a compact clump of eight red supergiants and four other candidates at some distance, all of them spectroscopically confirmed red supergiants. The majority of these objects must form an open cluster, which we name Alicante 10. Because of the high reddening and strong field contamination, the cluster sequence is not clearly seen in 2MASS or GPS-UKIDSS. From the observed sources, we derive E(J − KS) = 2.6 and d ≈ 6 kpc. Conclusions. Although the cluster is smaller than RSGC3, it has an initial mass in excess of 10 000 M⊙, and it seems to be part of the RSGC3 complex. With the new members this association already has 35 spectroscopically confirmed red supergiants, confirming its place as one of the most active sites of recent stellar formation in the Galaxy.

NGC 7419 as a template for red supergiant clusters

Context. The open cluster NGC 7419 is known to contain five red supergiants and a very high number of Be stars. However, there are conflicting reports about its age and distance that prevent a useful comparison with other clusters. Aims. We intend to obtain more accurate parameters for NGC 7419, using techniques different from those of previous authors, so that it may be used as a calibrator for more obscured clusters. Methods. We obtained Strömgren photometry of the open cluster NGC 7419, as well as classification spectroscopy of ~20 stars in the area. We then applied standard analysis and classification techniques. Results. We find a distance of 4 ± 0.4 kpc and an age of 14 ± 2 Myr for NGC 7419. The main-sequence turn-off is found at spectral type B1, in excellent agreement. We identify 179 B-type members, implying that there are more than 1200 M⊙ in B stars at present. Extrapolating this to lower masses indicates an initial cluster mass of between 7000 and 10 000 M⊙, depending on the shape of the initial mass function. We find a very high fraction (≈40%) of Be stars around the turn-off, but very few Be stars at lower masses. We also report for the first time a strong variability in the emission characteristics of Be stars. We verified that the parameters of the red supergiant members can be used to obtain accurate cluster parameters. Conclusions. NGC 7419 is sufficiently massive to serve as a testbed for theoretical predictions and as a template to compare more obscured clusters. The distribution of stars above the main-sequence turn-off is difficult to accommodate with current evolutionary tracks. Though the presence of five red supergiants is marginally consistent with theoretical expectations, the high number of Be stars and very low number of luminous evolved B stars hint at some unknown physical factor that is not considered in current synthesis models.

Single-cell derived clones from human adipose stem cells present different immunomodulatory properties

Human adipose mesenchymal stem cells are a heterogeneous population, where cell cultures derived from single cell-expanded clones present varying degrees of differential plasticity. This work focuses on the immunomodulatory/anti-inflammatory properties of these cells. To this end, 5 single cell clones were isolated (generally called 1.X and 3.X) from 2 volunteers. Regarding the expression level of the lineage-characteristic surface antigens, clones 1.10 and 1.22 expressed the lowest amounts, while clones 3.10 and 3.5 expressed more CD105 than the rest and clone 1.7 expressed higher amounts of CD73 and CD44. Regarding cytokine secretion, all clones were capable of spontaneously releasing high levels of IL-6 and low to moderate levels of IL-8. These differences can be explained in part by the distinct methylation profile exhibited by the clones. Furthermore and after lipopolysaccharide stimulation, clone 3.X produced the highest amounts of pro-inflammatory cytokines such as IL-1β, while clones 1.10 and 1.22 highly expressed IL-4 and IL-5. In co-culture experiments, clones 1.X are altogether more potent inhibitors than clones 3.X for proliferation of total, CD3+T, CD4+T and CD8+T lymphocytes and NK cells. The results of this work indicates that adipose stem cell population is heterogeneous in cytokine production profile, and that isolation, characterization and selection of the appropriate cell clone is a more exact method for the possible treatment of different patients or pathologies.

Characterization of a new murine retinal cell line (MU-PH1) with glial, progenitor and photoreceptor characteristics

Unlike fish and amphibians, mammals do not regenerate retinal neurons throughout life. However, neurogenic potential may be conserved in adult mammal retina and it is necessary to identify the factors that regulate retinal progenitor cells (RPC) proliferative capacity to scope their therapeutic potential. Müller cells can be progenitors for retinal neuronal cells and can play an essential role in the restoration of visual function after retinal injury. Some members of the Toll-like receptor (TLR) family, TLR2, TLR3 and TLR4, are related to progenitor cells proliferation. Müller cells are important in retinal regeneration and stable cell lines are useful for the study of retinal stem cell biology. Our purpose was to obtain a Müller-derived cell line with progenitor characteristics and potential interest in regeneration processes. We obtained and characterized a murine Müller-derived cell line (MU-PH1), which proliferates indefinitely in vitro. Our results show that (i) MU-PH1 cells expresses the Müller cell markers Vimentin, S-100, glutamine synthetase and the progenitor and stem cell markers Nestin, Abcg2, Ascl1, α-tubulin and β-III-tubulin, whereas lacks the expression of CRALBP, GFAP, Chx10, Pax6 and Notch1 markers; (ii) MU-PH1 cell line stably express the photoreceptor markers recoverin, transducin, rhodopsin, blue and red/green opsins and also melanopsin; (iii) the presence of opsins was confirmed by the recording of intracellular free calcium levels during light stimulation; (iv) MU-PH1 cell line also expresses the melatonin MT1 and MT2 receptors; (v) MU-PH1 cells express TLR1, 2, 4 and 6 mRNA; (vi) MU-PH1 express TLR2 at cell surface level; (vii) Candida albicans increases TLR2 and TLR6 mRNA expression; (viii) C. albicans or TLR selective agonists (Pam(3)CysSK(4), LPS) did not elicit morphological changes nor TNF-α secretion; (ix) C. albicans and Pam(3)CysSK(4) augmented MU-PH1 neurospheres formation in a statistically significant manner. Our results indicate that MU-PH1 cell line could be of great interest both as a photoreceptor model and in retinal regeneration approaches and that TLR2 may also play a role in retinal cell proliferation.