To analyze tear protein variations in patients suffering from dry eye symptoms in the presence of tear film instability but without epithelial defects.
Five microlitres of non-stimulated tears from ...60 patients, suffering from evaporative dry eye (EDE) with a break-up time (BUT) <10 s, and from 30 healthy subjects as control (no symptoms, BUT >10 s) were collected. Tear proteins were separated by mono and bi-dimensional SDS-PAGE electrophoresis and characterized by immunoblotting and enzymatic digestion. Digested peptides were analyzed by liquid chromatography coupled to electrospray ionization quadrupole-time of flight mass spectrometry followed by comparative data analysis into Swiss-Prot human protein database using Mascot. Statistical analysis were performed by applying a t-test for independent data and a Mann-Whitney test for unpaired data (P<0.05).
In EDE patients vscontrols, a significant decrease in levels of lactoferrin (data in %+/-SD): 20.15+/-2.64 vs 24.56+/-3.46 (P=0.001), lipocalin-1: 14.98+/-2.70 vs 17.73+/-2.96 (P=0.0001), and lipophilin A-C: 2.89+/-1.06 vs 3.63+/-1.37 (P=0.006) was revealed, while a significant increase was observed for serum albumin: 9.45+/-1.87 vs 3.46+/-1.87 (P=0.0001). No changes for lysozyme and zinc alpha-2 glycoprotein (P=0.07 and 0.7, respectively) were shown. Proteomic analysis showed a downregulation of lipophilin A and C and lipocalin-1 in patients, which is suggested to be associated with post-translational modifications.
Data show that tear protein changes anticipate the onset of more extensive clinical signs in early stage dry eye disease.
A progressive reduction of tetragonal distortion, heat of transition, Curie temperature, and relative dielectric constant has been observed on dense BaTiO3 ceramics with grain size decreasing from ...1200 to 50 nm. The correlations between grain size, extent of tetragonal distortion, and ferroelectric properties strongly support the existence of an intrinsic size effect. From the experimental trends the critical size for disappearance of ferroelectricity has been evaluated to be 10-30 nm. The strong depression of the relative permittivity observed for the nanocrystalline ceramics can be ascribed to the combination of the intrinsic size effect and of the size-dependent "dilution" effect of a grain boundary "dead" layer.
In this work, we show a new procedure to obtain an optimum DC polarisation potential that allows achieving the maximum possible sensitivity for analyte determination, when electrochemical techniques ...are used. This is valid whether purely DC or AC potentials are used for system polarisation. In addition, the optimum potential is lower than the ones generally used by the classical methods, avoiding redox-active interferents and rapid degradation of the working electrode. In our case, we obtain a reduction of 16% of the DC applied potential and an increase among 3 and 5 times in the sensitivity.
Graphical abstract
We have previously shown that the d16HER2 splice variant is linked to HER2-positive breast cancer (BC) tumorigenesis, progression and response to Trastuzumab. However, the mechanisms by which d16HER2 ...contributes to HER2-driven aggressiveness and targeted therapy susceptibility remain uncertain. Here, we report that the d16HER2-positive mammary tumor cell lines MI6 and MI7, derived from spontaneous lesions of d16HER2 transgenic (tg) mice and resembling the aggressive features of primary lesions, are enriched in the expression of Wnt, Notch and epithelial-mesenchymal transition pathways related genes compared with full-length wild-type (WT) HER2-positive cells (WTHER2_1 and WTHER2_2) derived from spontaneous tumors arising in WTHER2 tg mice. MI6 cells exhibited increased resistance to anoikis and significantly higher mammosphere-forming efficiency (MFE) and self-renewal capability than the WTHER2-positive counterpart. Furthermore, d16HER2-positive tumor cells expressed a higher fraction of CD29
/CD24
/SCA1
cells and displayed greater in vivo tumor engraftment in serial dilution conditions than WTHER2_1 cells. Accordingly, NOTCH inhibitors impaired mammosphere formation only in MI6 cells. A comparative analysis of stemness-related features driven by d16HER2 and WTHER2 in ad hoc engineered human BC cells (MCF7 and T47D) revealed a higher MFE and aldehyde dehydrogenase-positive staining in d16HER2- vs WTHER2-infected cells, sustaining consistent BC-initiating cell enrichment in the human setting. Moreover, marked CD44 expression was found in MCF7_d16 and T47D_d16 cells vs their WTHER2 and Mock counterparts. Clinically, BC cases from two distinct HER2-positive cohorts characterized by high levels of expression of the activated-d16HER2 metagene were significantly enriched in the Notch family and signal transducer genes vs those with low levels of the metagene.
Nannochloropsis oceanica is a microalga, highly concentrated in protein and EPA (eicosapentaenoic acid). It has a recalcitrant cell wall that decreases nutrient digestibility. The objective of this ...study is to evaluate the effect of Nannochloropsis oceanica feed inclusion on the muscle proteome of fattening rabbits.
Twenty rabbits were housed in individual cages for 5 weeks and were fed a control diet (n = 10) and one with 4.45% inclusion of microalga, replacing whole soybean meal (n = 10). After slaughter, samples of gastrocnemius muscle were taken for label-free proteomic analysis.
A total of 1497 proteins were identified, 46 with differential abundance. Control rabbits had high abundance of proteins related to protein metabolism, suggesting higher muscle protein turnover. They also had higher abundance of structural proteins, suggesting a less tender meat by comparison with algae-fed rabbits. These had high abundance of proteins related to amino acid catabolism (Phe, Tyr) and synthesis (Gln). In addition, they had high abundance of proteins related to protein breakdown, overall suggesting that more tender meat may result from algae feeding.
Microalgae are interesting feedstuffs particularly from a sustainability point of view. Their use in farm animal diets could simultaneously enhance the nutritional value and quality of meat. However, the biochemical pathways behind such improvement remain uncharacterized, namely in the rabbit species. By using a proteomics approach, this study will enlighten such biochemical pathways and relate them with productive performance parameters. In addition, results reflect some anti-nutritional properties of the recalcitrant cell wall over amino acid digestibility and consequently, its availability for protein synthesis.
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•Nannochloropsis oceanica feed inclusion does not affect animal performance.•Control rabbits have higher abundance of structural proteins in the muscle.•Dietary Nannochloropsis oceanica seems to affect amino acid metabolism in the muscle of rabbits.
Dense nanocrystalline BaTiO
3 ceramics with grain size (GS) down to 50
nm were studied by X-ray diffraction (XRD), differential scanning calorimetry (DSC), impedance spectroscopy and Raman ...spectroscopy. A continuous reduction of the tetragonal distortion towards the pseudo-cubic state was obtained when the GS was reduced. Therefore, even the finest structure (ceramic with average GS of 50
nm) is still non-centrosymmetric. The dielectric constant (
K) shows relative thermal stability in a large range of temperatures and is strongly depressed in the nanocrystalline ceramics, in comparison with the micrometric ones (
K being below 1000 for the ceramic with 50
nm GS). The losses are smaller than 5% in the frequency range of 10
2–10
6
Hz and temperatures below 200
°C. As the GS decreases, the structural phase transitions assume a more diffuse character. A decrease of the Curie temperature with reducing the GS was confirmed by X-ray, calorimetric and permittivity data. The Raman spectra collected for the range 80–800
K provided evidence for the presence of all the crystalline phases of BaTiO
3, as in single-crystal and micrometric ceramics; a few differences can be attributed to GS effects and to the high density of the non-ferroelectric grain boundaries. Evidence for the different phase transitions were provided by the disappearance of some bands and by anomalies in positions and intensities of selected Raman modes. The overall properties of the nanocrystalline BaTiO
3 ceramics can be explained as a combination of intrinsic effects, associated with the decrease of tetragonality and heat of transition with reducing GS, and extrinsic contributions due to the non-ferroelectric grain boundaries causing a “dilution” of the ferroelectric properties.
Theoretical models for small ferroelectric particles predict a progressive decrease of the Curie temperature, spontaneous lattice strain, and polarization until the critical size corresponding to ...transition to the cubic phase and disappearance of ferroelectricity is reached. In contrast, the behavior of nanocrystalline BaTiO3 ceramics with a grain size of approximate to 30 nm is dominated by extrinsic effects related to the grain boundaries which mask the expected downscaling of properties. While the noncubic crystal structure, the high dielectric constant (approximate to 1600) and the variation of permittivity with temperature suggest a ferroelectric behavior, very slim, and nearly linear polarization hysteresis loops are observed. Evidence for the existence of a ferroelectric domain structure with domains extending over several grains and of polarization switching at local scale is given by piezoresponse force microscopy. The suppression of macroscopic ferroelectric hysteresis and switching originates from a frozen domain structure stable under an external field owing to the effects exerted by the grain boundaries, such as the clamping of the domain walls and the hindrance of polarization switching. Furthermore, the depolarization field originated by the low-permittivity nonferroelectric grain boundaries can cause a significant reduction of polarization. If the grain size is small enough, the ceramic is expected to undergo a "phase transition" to a polar phase with nonswitchable polarization. The BaTiO3 ceramics with grain size of 30 nm investigated in the present study are deemed to be close to this transition.
•High frequency dielectric relaxation process is observed and interpreted.•Multiple conductivity processes analyzed by fractal Jonscher’s relationship.•Calculated conductivity activation energies ...fall in 0.30–0.80eV range.
The behavior of three xBaTiO3–(1−x)Ni0.5Zn0.5Fe2O4 composites (x=0.50, 0.60 and 0.70M ratios) were studied using the broadband dielectric spectroscopy methods in the broad frequency and temperature ranges. x=0.70 composition clearly exhibits similar dielectric response to that of pure barium titanate, while the response of the composite with the highest concentration of nickel–zinc ferrite is largely influenced by the conductivity, especially at the highest temperatures and lowest frequencies. In the non-dispersive region the permittivity gradually increases with increasing level of high-permittivity barium titanate material as a result of the “sum property”, while the low frequency behavior described by complex contributions do not show a systematical modification with the changing composition. At around 108–1010Hz frequencies the dispersion process was observed and approximated with the Cole–Cole equation, finding the parameters describing the process. Presumably, lattices of barium titanate in close vicinity to nickel–zinc ferrite could lead to the distortion and some shifting of ions in barium titanate lattice, possibly affecting the states that Ti can occupy. The conductivity process was approximated with the recently suggested generalized Joncher’s relationship, which manages to describe both low and high frequency conductivity processes observed. The computed corresponding activation energies for all the samples were found to be similar to oxide composites containing ferrites with ionic conduction contributions.
CD49d, the alpha-chain of the integrin heterodimer α4β1, was identified among the strongest predictors of overall survival (OS) in chronic lymphocytic leukemia (CLL), along with IGHV mutational ...status and deletion of the 17p chromosome involving TP53. In addition to TP53, the clinical relevance of NOTCH1, SF3B1 and BIRC3 gene mutations has been recently emphasized. By analyzing a cohort of 778 unselected CLL patients, we assessed the clinical relevance of CD49d as an OS predictor in subgroups defined by mutation/deletion of the TP53, NOTCH1, SF3B1 and BIRC3 genes. In this context, CD49d emerged as an independent predictor of OS in multivariate Cox analysis (Hazard ratio =1.88, P<0.0001). Consistently, high CD49d expression identified CLL subsets with inferior OS in the context of each category of a previously reported hierarchical risk stratification model. Moreover, by evaluating the relative importance of biological prognosticators by random survival forests, CD49d was selected among the top-ranked OS predictor (variable importance =0.0410), along with IGHV mutational status and TP53 abnormalities. These results confirmed CD49d as an independent negative OS prognosticator in CLL also in comprehensive models comprising the novel recurrent mutations. In this context, TP53 disruption and NOTCH1 mutations retained prognostic relevance, in keeping with their roles in CLL cell immuno-chemoresistance.
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