Heat Shock Alters the Proteomic Profile of Equine Mesenchymal Stem Cells

The aim of this research was to determine the impact of heat stress on cell differentiation in an equine mesenchymal stem cell model (EMSC) through the application of heat stress to primary EMSCs as they progressed through the cell specialization process. A proteomic analysis was performed using mass spectrometry to compare relative protein abundances among the proteomes of three cell types: progenitor EMSCs and differentiated osteoblasts and adipocytes, maintained at 37 °C and 42 °C during the process of cell differentiation. A cell-type and temperature-specific response to heat stress was observed, and many of the specific differentially expressed proteins were involved in cell-signaling pathways such as Notch and Wnt signaling, which are known to regulate cellular development. Furthermore, cytoskeletal proteins profilin, DSTN, SPECC1, and DAAM2 showed increased protein levels in osteoblasts differentiated at 42 °C as compared with 37 °C, and these cells, while they appeared to accumulate calcium, did not organize into a whorl agglomerate as is typically seen at physiological temperatures. This altered proteome composition observed suggests that heat stress could have long-term impacts on cellular development. We propose that this in vitro stem cell culture model of cell differentiation is useful for investigating molecular mechanisms that impact cell development in response to stressors.     

diaPASEF Proteomics and Feature Selection for the Description of Sputum Proteome Profiles in a Cohort of Different Subtypes of Lung Cancer Patients and Controls

The high mortality, the presence of an initial asymptomatic stage and the fact that diagnosis in early stages reduces mortality justify the implementation of screening programs in the populations at risk of lung cancer. It is imperative to develop less aggressive methods that can complement existing diagnosis technologies. In this study, we aimed to identify lung cancer protein biomarkers and pathways affected in sputum samples, using the recently developed diaPASEF mass spectrometry (MS) acquisition mode. The sputum proteome of lung cancer cases and controls was analyzed through nano-HPLC–MS using the diaPASEF mode. For functional analysis, the results from differential expression analysis were further analyzed in the STRING platform, and feature selection was performed using sparse partial least squares discriminant analysis (sPLS-DA). Our results showed an activation of inflammation, with an alteration of pathways and processes related to acute-phase, complement, and immune responses. The resulting sPLS-DA model separated between case and control groups with high levels of sensitivity and specificity. In conclusion, we showed how new-generation proteomics can be used to detect potential biomarkers in sputum samples, and ultimately to discriminate patients from controls and even to help to differentiate between different cancer subtypes.     

Somatostatin Receptor Splicing Variant sst5TMD4 Overexpression in Glioblastoma Is Associated with Poor Survival,Increased Aggressiveness Features,and Somatostatin Analogs Resistance

Glioblastoma (GBM) is the most malignant and lethal brain tumor. Current standard treatment consists of surgery followed by radiotherapy/chemotherapy; however, this is only a palliative approach with a mean post-operative survival of scarcely ~12–15 months. Thus, the identification of novel therapeutic targets to treat this devastating pathology is urgently needed. In this context, the truncated splicing variant of the somatostatin receptor subtype 5 (sst5TMD4), which is produced by aberrant alternative splicing, has been demonstrated to be overexpressed and associated with increased aggressiveness features in several tumors. However, the presence, functional role, and associated molecular mechanisms of sst5TMD4 in GBM have not been yet explored. Therefore, we performed a comprehensive analysis to characterize the expression and pathophysiological role of sst5TMD4 in human GBM. sst5TMD4 was significantly overexpressed (at mRNA and protein levels) in human GBM tissue compared to non-tumor (control) brain tissue. Remarkably, sst5TMD4 expression was significantly associated with poor overall survival and recurrent tumors in GBM patients. Moreover, in vitro sst5TMD4 overexpression (by specific plasmid) increased, whereas sst5TMD4 silencing (by specific siRNA) decreased, key malignant features (i.e., proliferation and migration capacity) of GBM cells (U-87 MG/U-118 MG models). Furthermore, sst5TMD4 overexpression in GBM cells altered the activity of multiple key signaling pathways associated with tumor aggressiveness/progression (AKT/JAK-STAT/NF-κB/TGF-β), and its silencing sensitized GBM cells to the antitumor effect of pasireotide (a somatostatin analog). Altogether, these results demonstrate that sst5TMD4 is overexpressed and associated with enhanced malignancy features in human GBMs and reveal its potential utility as a novel diagnostic/prognostic biomarker and putative therapeutic target in GBMs.     

The steryl ester and phospholipid fatty acids of the spongeAgelas conifera from the Colombian Caribbean

The steryl ester and phospholipid fractions of the marine spongeAgelas conifera were isolated and analyzed. The fatty acyl components of the steryl ester and phospholipid fractions as determined by gas chromatography and gas chromatography/mass spectrometry were very similar and consisted of 56.8 and 62.7% of C₁₄−C₂₀ acids (normal; branched, especiallyiso andanteiso; and monounsaturated, particularly Δ⁹ and Δ¹¹ acids) and of 43.1 and 35.5% of C₂₄−C₂₆ acids (Δ^5,9 diunsaturated acids), respectively. The major constituent fatty acids detected were 13-methyltetradecanoic,n-hexadecanoic, 10-methylhexadecanoic, 11-octadecenoic, 12-methyloctadecanoic, 5,9-pentacosadienoic and 5,9-hexacosadienoic acids. The phospholipids isolated were identified as phosphatidylcholine (37%), phosphatidylserine (34%), phosphatidylethanolamine (16%) and phosphatidylinositol (11%). The distribution of fatty acids within the phospholipid classes was also determined.     

Fullerene C₆₀ as a multifunctional system for drug and gene delivery

The fullerene family, and especially C(60), has delighted the scientific community during the last 25 years with perspective applications in a wide variety of fields, including the biological and the biomedical domains. Several biomedical uses have been explored using water-soluble C(60)-derivatives. However, the employment of fullerenes for drug delivery is still at an early stage of development. The design and synthesis of multifunctionalized and multimodal C(60) systems able to cross the cell membranes and efficiently deliver active molecules is an attracting challenge that involves multidisciplinary strategies. Promising results have emerged in the last years, bringing fullerenes again to the front of interest. Herein, the state of the art of this emerging field is presented and illustrated with some of the most representative examples.     

Phase and Rheological Behavior of Cetyldimethylbenzylammonium Salicylate (CDBAS) and Water

The temperature–composition phase diagram in the diluted region of the cationic surfactant cetyldimethylbenzylammonium salicylate/water system was studied with a battery of techniques. The Krafft temperature (T _k = 33 ± 1 °C) was measured by differential scanning calorimetry, polarizing microscopy, conductimetry, viscosimetry, and rheometry. The critical vesicle concentration (cvc, ~0.002 wt%) and a vesicle–micellar transition (cvm, ~0.005 wt%) was detected at a temperature of 35 °C. Below T _k and concentrations ≤2 wt%, a transparent solution is formed (I). Above 2–8.5 wt%, a lamellar (L₁) phase forms. At higher concentrations and up to 12 wt%, a second lamellar phase (L₂) is detected. From 12.4 to 15.5 wt%, an emulsion phase (E) is formed. Rheological dynamic measurements for the I phase indicate that the system exhibits a predominantly viscous behavior (G′ < G″) for concentrations lower than the overlap or entanglement concentration (C _e, ~0.75 wt%). At higher concentrations, wormlike micelles form and the elastic behavior predominates (G′ > G″). The elastic (G′) modulus collapses in a concentration–time master curve in the whole reduced frequencies range ωτ _c examined, whereas the viscous modulus (G″) collapses only at reduced frequencies lower than 0.1. Reduced stress plotted as a function of the reduced shear rate yields a good superposition of the curves at the different concentrations up to the onset of the non-linear behavior.     

Mechanical properties of carbon nanofiber/fiber-reinforced hierarchical polymer composites manufactured with multiscale-reinforcement fabrics

Hierarchical polymer composites – defined as carbon nanofiber/fiber-reinforced polymer composites – were manufactured using multiscale-reinforcement fabrics (MRFs) and they were characterized for their mechanical properties. The MRFs were fabricated by electrophoretic deposition of carboxylic acid- or amine-functionalized carbon nanofibers (CNFs) on the surface of sized or unsized carbon fiber layers. Compared to the base composite (not containing CNFs), the hierarchical composites containing the functionalized CNFs showed an increase in interlaminar shear strength (ILSS) and compressive strength. Panels containing amine-functionalized CNFs had the highest increase in properties: 12% in ILSS and 13% in compressive strength. The reinforcement mechanism was also investigated with emphasis placed on the fiber/matrix interface and the load transfer between matrix, CNFs, and carbon fiber.     

Oxygen migration on the graphene surface. 2. Thermochemistry of basal-plane diffusion (hopping)

Thermodynamic affinities, activation energies and diffusion coefficients for oxygen mobility on the graphene surface are calculated using density functional theory (DFT). We report and discuss the effects of geometry, charge distribution and heteroatom substitution on the migration of epoxy oxygen on the basal plane: both the driving force and the ease of surface hopping are very sensitive to their variations. A significant decrease in the hopping energy barrier is observed when graphene contains free edge sites and oxygen functionalities, as well as upon an increase in electron density; conversely, the barrier increases as a consequence of electron removal, and the propensity for graphene ‘unzipping’ also increases. There is a correlation between the hopping barrier and the C–O bond strength of the leaving epoxide group. Under the most favorable conditions investigated, oxygen mobility is quite high, of the same order as that of gas-phase O₂ in micropores (ca. 10⁻⁹ m²/s). This is consistent with the increasingly acknowledged role of basal-plane oxygen as a protagonist (e.g., reaction intermediate), instead of a spectator, in the wide variety of adsorption and reaction processes involving sp²-hybridized carbon materials.     

Fast and efficient synthesis of high molecular weight poly(epsilon‐caprolactone) diols by microwave‐assisted polymer synthesis

Because of advantageous features such as shorter reaction times, greater yields, limited generation of by‐products and relatively easy and straightforward scale‐up, microwave‐assisted synthesis has become a very appealing tool in organic synthesis. Conversely, its implementation in the context of the synthesis of biomaterials for biopharmaceutical applications has been more limited. The present work reports on the fast and efficient microwave‐assisted synthesis of poly(ethylene glycol) (PEG)‐initiated poly(ε‐caprolactone) diols (PCL) by the ring‐opening polymerization (ROP) of ε‐caprolactone using stannous octanoate as catalyst. Since the PEG content in the synthesized copolymers was extremely low (0.2–1.9%), products were highly hydrophobic and displayed the intrinsic thermal properties of pure PCL. As opposed to the more time‐consuming conventional thermally‐driven synthesis that usually demands 2–3 h, the microwave technique resulted in intermediate to high molecular weight PEG‐PCL derivatives within 10–15 min. The influence of different parameters affecting the synthetic process, namely monomer‐to‐initiator ratio, reaction time, catalyst concentration and the presence, type, and concentration of solvent were thoroughly investigated. © 2011 Wiley Periodicals, Inc. J Appl Polym Sci, 2011     

Purity,quality and stability of Argentinean virgin olive oils

In this paper we evaluate the stability, purity and regulated quality composition of fatty acids and sterols (both physico‐chemical and sensory) of commercial Argentinean virgin olive oils in order to evaluate their acceptance on the world market. For this purpose, samples of the best known and most widely distributed oils in supermarkets located in Buenos Aires (Argentina) were acquired. After thoroughly analysing these samples, only 20% were considered to have an acceptable quality. However, some were excluded because of their high campesterol content, which could be an intrinsic characteristic of these oils. The most useful analytical parameter used to confirm authenticity was ECN‐42 R – ECN‐42 T, followed by wax content and 3.5 stigmastadienes. Only 24% of the extra‐virgin olive oil samples were classified as ‘extra‐virgin’ from the regulated quality viewpoint. The low oleic and high linolenic acid contents of the Argentinean virgin olive oils stand out when compared with European virgin olive oils. The oxidative stability values may be considered very low, indeed even lower than those obtained in Spanish virgin olive oils.