HIF-1α and Pro-Inflammatory Signaling Improves the Immunomodulatory Activity of MSC-Derived Extracellular Vesicles

Despite the strong evidence for the immunomodulatory activity of mesenchymal stromal cells (MSCs), clinical trials have so far failed to clearly show benefit, likely reflecting methodological shortcomings and lack of standardization. MSC-mediated tissue repair is commonly believed to occur in a paracrine manner, and it has been stated that extracellular vesicles (EVs) secreted by MSCs (EVMSC) are able to recapitulate the immunosuppressive properties of parental cells. As a next step, clinical trials to corroborate preclinical studies should be performed. However, effective dose in large mammals, including humans, is quite high and EVs industrial production is hindered by the proliferative senescence that affects MSCs during massive cell expansion. We generated a genetically modified MSC cell line overexpressing hypoxia-inducible factor 1-alpha and telomerase to increase the therapeutic potency of EVMSC and facilitate their large-scale production. We also developed a cytokine-based preconditioning culture medium to prime the immunomodulatory response of secreted EVs (EV_MSC-T-HIF^c). We tested the efficacy of this system in vitro and in a delayed-type hypersensitivity mouse model. MSC-T with an HIF-1α-GFP lentiviral vector (MSC-T-HIF) can be effectively expanded to obtain large amounts of EVs without major changes in cell phenotype and EVs composition. EV_MSC-T-HIF^c suppressed the proliferation of activated T-cells more effectively than did EVs from unmodified MSC in vitro, and significantly blunted the ear-swelling response in vivo by inhibiting cell infiltration and improving tissue integrity. We have developed a long-lived EV source that secretes high quantities of immunosuppressive EVs, facilitating a more standard and cost-effective therapeutic product.     

High efficiency and reusability of iridium complexes adsorbed in montmorillonite clay on catalytic hydrogenation of imines

Heterogenised homogeneous catalytic hydrogenation of aldimines by removable and reusable immobilised iridium complexes on montmorillonite clay. This revised version was published online in July 2006 with corrections to the Cover Date.     

Using Structural Equation Modeling (SEM) for the Study of Impurity Profiles of Drug Substances

The impurity profile is one of the most important quality characteristics of a drug substance. Although it is always desirable to determine the chemical structure of all impurities forming the impurity profile, unfortunately this is not always economically and technically feasible because of the extremely low concentrations at which some impurities may be found in the drug substance. Therefore, alternative approaches to the chemical analysis are needed for trying to determine the origin of the unidentified impurities.

In a previous study conducted by our group, based on exploratory (principal component and hierarchical cluster) analysis, we were able to suggest a hypothesis for explaining the origin of the unidentified impurities of a drug substance. However, there was still a concern that alternative hypotheses might explain the same phenomenon equally well. This article explores the application of recent developments in structural equation modeling for the systematic generation and selection of hypotheses (models) worthy of being confirmed by chemical research.     

Dual holographic interferometry for measuring the three velocity components in a fluid plane

A technique that allows one to measure simultaneously the three velocity components in a fluid plane is presented. One obtains the quantitative information from only one holographic recording by combining two different reconstruction processes. As both processes use an interferometric comparison of two waves, we refer to this technique as dual holographic interferometry. The far-field fringe pattern that is obtained when reconstruction is made with an expanded laser beam allows one to determine the in-plane velocity components. The image-field fringe pattern that is obtained when a pointwise laser beam is used for reconstruction contains information about an out-of-plane velocity component. As the two reconstruction processes have different sensitivities, two different ways to combine them are proposed. The system has been demonstrated in a fluidlike solid object and in a convective flow.     

MR imaging of baker cysts—prevalence and relation to internal derangements of the knee

Rationale and objectives: To evaluate the prevalence of popliteal cysts in patients studied with magnetic resonance (MR) imaging. Cyst presence and volume will be related to other internal derangement of the knee.Materials and methods: Three hundred and eighty-two consecutive patients with a MR study of the knee were included. Images were obtained in the three spatial orthogonal planes and evaluated through consensus. Four categories were established for the Baker cyst and synovial fluid (absence, minimum, moderate and massive), and the presence of meniscal, cruciate ligaments and cartilage lesions were recorded. Statistical analysis was carried out with bivariate analysis (χ² of Pearson and Gamma tests).Results: From the 382 patients, 145 had Baker cysts (38.0%). Cyst content was minimum in 99, moderate in 34 and massive in 12. Joint effusion was observed in 269 patients (70.4%), being minimal in 140 patients, moderate in 119 and massive in ten. Meniscal lesions were observed in 195 patients (51%), while 58 patients (15%) had a cruciate ligament lesion. Baker cyst has a statistically significant direct relationship with the presence and quantity of synovial fluid (P=0.002) and with the presence and type of meniscal lesion (P=0.01) but not with cruciate ligaments or cartilage lesions.Conclusions: The prevalence of Baker cysts in MR studies of the knee is high. Its presence and volume are related to the quantity of synovial fluid, and to the presence and severity of meniscal lesions.     

An approach for automatic generation of adaptive hypermedia in education with multilingual knowledge discovery techniques

This work describes a framework that combines techniques from Adaptive Hypermedia and Natural Language processing in order to create, in a fully automated way, on-line information systems from linear texts in electronic format, such as textbooks. The process is divided into two steps: an off-line processing step, which analyses the source text, and an on-line step, which executes when a user connects to the system with a web browser, moment at which the contents and hyperlinks are generated. The framework has been implemented as the Welkin system, which has been used to build three adaptive on-line information sites in a quick and easy way. Some controlled experiments have been performed with real users aimed to provide positive feedback on the implementation of the system.     

Asymptotic normality of the Nadaraya–Watson semivariogram estimators

In this work, the Nadaraya–Watson semivariogram estimation is considered for both the isotropic and the anisotropic settings. Several properties of these estimators are analyzed and, particularly, their asymptotic normality is established in terms of unknown characteristics of the random process. The latter provides a theoretical procedure for construction of confidence intervals for the semivariogram via the normal quantiles, which in practice must be appropriately estimated. A numerical study is included to illustrate the performance of the Nadaraya–Watson estimation when used to obtain confidence intervals.      

CHARACTERIZATION OF EVA-BASED ADHESIVES CONTAINING DIFFERENT AMOUNTS OF ROSIN ESTER OR POLYTERPENE TACKIFIER

Different amounts (50-170 php--parts per hundred parts of EVA, 33-63 wt%) of two tackifiers (hydrogenated rosin ester, polyterpene resin) were added to an ethylene vinyl acetate (EVA) copolymer containing 28 wt% vinyl acetate. The EVA and the tackifier were characterized using infrared (IR) spectroscopy, DSC measurements, and stress-controlled plate-plate rheology. The properties and compatibility of the EVA-tackifier mixtures were studied using DSC, DMTA, and stress-controlled plate-plate rheology. Immediate adhesion was measured as a quantification of tack, and the T-peel strength of roughened styrene-butadiene rubber/EVA-tackifier adhesive joints was also obtained. The increase in the amount of tackifier noticeably changed the crystallinity of polyethylene blocks in the EVA, and the temperature at the cross-over between the curves of the storage and loss moduli as a function of the temperature was displaced to a lower value. Whereas the hydrogenated rosin ester was compatible with the amorphous ethylene vinyl acetate copolymer regions of the EVA (Tg value increased) reducing its crystallinity, the polyterpene resin was compatible with the polyethylene blocks of the EVA (T g value was not modified), increasing its crystallinity. Immediate adhesion of the EVA-tackifier mixtures was improved by adding both hydrogenated rosin ester and polyterpene tackifiers. On the other hand, there was an optimum tackifier content at which the maximum T-peel strength value was obtained.     

Highly Hydrophilic and Lipophilic Derivatives of Bile Salts

Lipophilicity of 15 derivatives of sodium cholate, defined by the octan-1-ol/water partition coefficient (log P), has been theoretically determined by the Virtual log P method. These derivatives bear highly hydrophobic or highly hydrophilic substituents at the C3 position of the steroid nucleus, being linked to it through an amide bond. The difference between the maximum value of log P and the minimum one is enlarged to 3.5. The partition coefficient and the critical micelle concentration (cmc) are tightly related by a double-logarithm relationship (VirtuallogP=−(1.00±0.09)log(cmcmM)+(2.79±0.09)), meaning that the Gibbs free energies for the transfer of a bile anion from water to either a micelle or to octan-1-ol differ by a constant. The equation also means that cmc can be used as a measurement of lipophilicity. The demicellization of the aggregates formed by three derivatives of sodium cholate bearing bulky hydrophobic substituents has been studied by surface tension and isothermal titration calorimetry. Aggregation numbers, enthalpies, free energies, entropies, and heat capacities, ΔC_P,demic, were obtained. ΔC_P,demic, being positive, means that the interior of the aggregates is hydrophobic.     

Studies on the interactions between bovine β-lactoglobulin and chitosan at the solid-liquid interface

Chitosan ultrathin films have been formed on polycrystalline Au substrates using the LbL technique with the purpose of studying its interaction with bovine β-lactoglobulin (β-LG) at the solid-liquid interface. The immobilization of chitosan was followed by Quartz Crystal Microbalance with energy dissipation (QCM-D), Cyclic Voltammetry (CV) and Electrochemical Impedance Spectroscopy (EIS). The behavior of the chitosan films in the presence of β-LG solutions with different bulk concentrations ([β-LG]), ionic strength (I), and pH has been investigated using the same techniques plus Atomic Force Microscopy (AFM). The results showed that for pHs lower than protein's pI, weak intermolecular forces (H bonding, Van der Waals, hydrophobic, etc.) are established between β-LG and chitosan (especially close to the pI) leading to low coverage nonspecific adsorption. On the contrary when pH > pI, strong ionic bonding through attractive electrostatic interactions lead to high coverage adsorbed phases composed of large β-LG aggregates. The adsorption process was shown to consist of a relatively fast step (in which these interactions are predominant) which is followed, once the β-LG monolayer is exceeded, by the slow formation of thicker and increasingly viscoelastic films through β-LG self-aggregation. QCM-D and AFM experiments unveiled the role of [β-LG] and I on the formation of these aggregates. The adsorption isotherm built from impedance data in the medium-low [β-LG] range (0.001-0.3 mg mL⁻¹), showed good fitting to the Langmuir model confirming that the formation of one β-LG monolayer is achieved in this concentration range.