Cells as factories for humanized encapsulation

Biocompatibility is of paramount importance for drug delivery, tumor labeling, and in vivo application of nanoscale bioprobes. Until now, biocompatible surface processing has typically relied on PEGylation and other surface coatings, which, however, cannot minimize clearance by macrophages or the renal system but may also increase the risk of chemical side effects. Cell membranes provide a generic and far more natural approach to the challenges of encapsulation and delivery in vivo. Here we harness for the first time living cells as "factories" to manufacture cell membrane capsules for encapsulation and delivery of drugs, nanoparticles, and other biolabels. Furthermore, we demonstrate that the built-in protein channels of the new capsules can be utilized for controlled release of encapsulated reagents.     

Digestibility by dairy cows of monosaccharide constituents in total mixed rations containing citrus pulp

Ten lactating cows were divided into two groups and individually fed ad libitum one of two experimental total mixed rations (TMR) as follows: 1) a TMR containing 20% corn grain and 10% dry citrus pulp (high corn); and 2) a TMR containing 21% citrus pulp and 9% corn grain (high citrus pulp). Both TMR also contained corn silage (28%), legume haylage (19.5%), extruded full fat soybeans (13.5 to 14%), soybean meal (6.5%), and minerals and vitamins. Voluntary dry matter intake of cows consuming the high corn ration tended to be higher than for cows in the high citrus pulp group (22.2 vs. 20.7 +/- 1.0 kg/cow per day, respectively). Digestibility of NDF was higher for the TMR with high citrus pulp compared with the TMR with higher corn. Across treatments, neutral detergent fiber (NDF) glucose was more digestible than NDF xylose and uronic acids and less digestible than NDF arabinose, galactose, and mannose. Digestibility of total neutral detergent soluble carbohydrates was also higher in the high citrus pulp group (86.8, high citrus pulp vs. 80.3 +/- 1.7%, high corn); with glucose and uronic acids accounting for most of that difference. Consequently, higher total carbohydrate digestibility was obtained in the TMR containing higher citrus pulp compared with the TMR with a higher percentage of corn (77.1 and 72.5 +/- 1.0%, respectively). Slightly lower carbohydrate intake in cows fed higher citrus pulp was compensated by higher digestibility of carbohydrates and protein. Thereby, partial replacement of corn by citrus pulp in TMR of high producing dairy cows improved feed efficiency.     

Unbalanced voltage faults: the impact on structural loads of doubly fed asynchronous generator wind turbines

This paper investigates the impact that unbalanced voltage faults have on wind turbine structural loads. In such cases, electromagnetic torque oscillations occur at two times the supply voltage frequency. The objectives of this work are to quantify wind turbine structural loads induced by unbalanced voltage faults relative to those during normal operation; and to evaluate the potential for reducing structural loads with the control of the generator. The method applied is integrated dynamic analysis. Namely, dynamic analysis with models that consider the most important aeroelastic, electrical, and control dynamics in an integrated simulation environment based on an aeroelastic code (HAWC2) and software for control design (Matlab/Simulink). In the present analysis, 1 Hz equivalent loads are used to compare fatigue loads, whereas maximum–minimum values are used to compare extreme loads. A control concept based on resonant filters demonstrates reduction of the structural loads (shaft torsion and tower top side‐to‐side moment) induced by an unbalanced voltage fault.Copyright © 2013 John Wiley & Sons, Ltd.     

Interaction of silicon-based quantum dots with gibel carp liver: oxidative and structural modifications

Quantum dots (QDs) interaction with living organisms is of central interest due to their various biological and medical applications. One of the most important mechanisms proposed for various silicon nanoparticle-mediated toxicity is oxidative stress. We investigated the basic processes of cellular damage by oxidative stress and tissue injury following QD accumulation in the gibel carp liver after intraperitoneal injection of a single dose of 2 mg/kg body weight Si/SiO₂ QDs after 1, 3, and 7 days from their administration.QDs gradual accumulation was highlighted by fluorescence microscopy, and subsequent histological changes in the hepatic tissue were noted. After 1 and 3 days, QD-treated fish showed an increased number of macrophage clusters and fibrosis, while hepatocyte basophilia and isolated hepatolytic microlesions were observed only after substantial QDs accumulation in the liver parenchyma, at 7 days after IP injection.Induction of oxidative stress in fish liver was revealed by the formation of malondialdehyde and advanced oxidation protein products, as well as a decrease in protein thiol groups and reduced glutathione levels. The liver enzymatic antioxidant defense was modulated to maintain the redox status in response to the changes initiated by Si/SiO₂ QDs. So, catalase and glutathione peroxidase activities were upregulated starting from the first day after injection, while the activity of superoxide dismutase increased only after 7 days. The oxidative damage that still occurred may impair the activity of more sensitive enzymes. A significant inhibition in glucose-6-phosphate dehydrogenase and glutathione-S-transferase activity was noted, while glutathione reductase remained unaltered.Taking into account that the reduced glutathione level had a deep decline and the level of lipid peroxidation products remained highly increased in the time interval we studied, it appears that the liver antioxidant defense of Carassius gibelio does not counteract the oxidative stress induced 7 days after silicon-based QDs exposure in an efficient manner.     

Galectin 1—A Key Player between Tissue Repair and Fibrosis

Galectins are ten family members of carbohydrate-binding proteins with a high affinity for β galactose-containing oligosaccharides. Galectin-1 (Gal-1) is the first protein discovered in the family, expressed in many sites under normal and pathological conditions. In the first part of the review article, we described recent advances in the Gal-1 modulatory role on wound healing, by focusing on the different phases triggered by Gal-1, such as inflammation, proliferation, tissue repair and re-epithelialization. On the contrary, Gal-1 persistent over-expression enhances angiogenesis and extracellular matrix (ECM) production via PI3K/Akt pathway activation and leads to keloid tissue. Therefore, the targeted Gal-1 modulation should be considered a method of choice to treat wound healing and avoid keloid formation. In the second part of the review article, we discuss studies clarifying the role of Gal-1 in the pathogenesis of proliferative diabetic retinopathy, liver, renal, pancreatic and pulmonary fibrosis. This evidence suggests that Gal-1 may become a biomarker for the diagnosis and prognosis of tissue fibrosis and a promising molecular target for the development of new and original therapeutic tools to treat fibrosis in different chronic diseases.     

Dopamine Photochemical Behaviour under UV Irradiation

To understand the photochemical behaviour of the polydopamine polymer in detail, one would also need to know the behaviour of its building blocks. The electronic absorption, as well as the fluorescence emission and excitation spectra of the dopamine were experimentally and theoretically investigated considering time-resolved fluorescence spectroscopy and first-principles quantum theory methods. The shape of the experimental absorption spectra obtained for different dopamine species with standard, zwitterionic, protonated, and deprotonated geometries was interpreted by considering the advanced equation-of-motion coupled-cluster theory of DLPNO-STEOM. Dynamical properties such as fluorescence lifetimes or quantum yield were also experimentally investigated and compared with theoretically predicted transition rates based on Fermi’s Golden Rule-like equation. The results show that the photochemical behaviour of dopamine is strongly dependent on the concentration of dopamine, whereas in the case of a high concentration, the zwitterionic form significantly affects the shape of the spectrum. On the other hand, the solvent pH is also a determining factor for the absorption, but especially for the fluorescence spectrum, where at lower pH (5.5), the protonated and, at higher pH (8.0), the deprotonated forms influence the shape of the spectra. Quantum yield measurements showed that, besides the radiative deactivation mechanism characterized by a relatively small QY value, non-radiative deactivation channels are very important in the relaxation process of the electronic excited states of different dopamine species.     

Blood Clots versus PRF: Activating TGF-β Signaling and Inhibiting Inflammation In Vitro

The preparation of platelet-rich fibrin (PRF) requires blood centrifugation to separate the yellow plasma from the red erythrocyte fraction. PRF membranes prepared from coagulated yellow plasma are then transferred to the defect sites to support tissue regeneration. During natural wound healing, however, it is the unfractionated blood clot (UBC) that fills the defect site. It is unclear whether centrifugation is necessary to prepare a blood-derived matrix that supports tissue regeneration. The aim of the present study was to compare lysates prepared from PRF and UBC based on bioassays and degradation of the respective membranes. We report here that lysates prepared from PRF and UBC membranes similarly activate TGF-β signaling, as indicated by the expression of interleukin 11 (IL-11), NADPH oxidase 4 (NOX-4) and proteoglycan 4 (PRG4) in gingival fibroblasts. Consistently, PRF and UBC lysates stimulated the phosphorylation and nuclear translocation of Smad3 in gingival fibroblasts. We further observed that PRF and UBC lysates have comparable anti-inflammatory activity, as shown by the reduction in lipopolysaccharide (LPS)-induced IL-6, inducible nitric oxidase synthase (iNOS) and cyclooxygenase 2 (COX-2) expression in RAW264.7 cells. Moreover, inflammation induced by Poly (1:C) HMW and FSL-1, which are agonists of Toll-like receptor (TLR) 3 and 2/6, respectively, was reduced by both PRF and UBC. PRF and UBC lysates reduced the nuclear translocation of p65 in LPS-induced RAW264.7 cells. In contrast to the similar activity observed in the bioassays, UBC membranes lack the structural integrity of PRF membranes, as indicated by the rapid and spontaneous disintegration of UBC membranes. We show here that the lysates prepared from PRF and UBC possess robust TGF-β and anti-inflammatory activity. However, visual inspection of the PRF and UBC membranes confirmed the negative impact of erythrocytes on the structural integrity of membranes prepared from whole blood. The data from the present study suggest that although both UBC and PRF have potent TGF-β and anti-inflammatory activity, UBC does not have the strength properties required to be used clinically to prepare applicable membranes. Thus, centrifugation is necessary to generate durable and clinically applicable blood-derived membranes.     

Impact of packaging properties on the physical-chemical-microbiological-sensory characteristics of Ricotta cheese during storage

Ricotta is a highly perishable cheese of which shelf-life is strongly influenced by the properties of packages in which it is stored. Four plastic packages currently in use in Italy (IT), Romania (RO), Slovenia (SLO), and Spain (SP) are characterized in term of structure, barrier, and antimicrobial properties, and their efficiency in the ricotta storage at 4°C is assessed. Differential scanning calorimetry and Fourier-transform infrared spectroscopy analysis indicate that the IT and SP packages are made of low-density polyethylene and the RO and SLO packages consist of polyethylene and polyamide 6. Permeability to the water vapour and oxygen transmission rate vary as follows: IT>SLO > RO > SP and SP > IT>RO > SLO, respectively. The SP package expressed the highest antimicrobial inhibition whilst the IT package the lowest. The maximum amounts of volatile compounds in cheese were reached after 3 days in the IT and SP packages and after 6 days in the SLO and RO packages. The score 2.8 for cheese overall acceptability was exceeded after 7 days in the IT, RO, and SP packages and 6 days in the SLO package. Barrier properties of packages significantly influence the shelf-life of ricotta. Low barrier properties resulted in high level of oxygen and water vapours inside the headspace of packages ultimately favour the microbial development. The simultaneous compliance of microbiological safety level and overall acceptability score gives ricotta a shelf-life of 3 days in the IT, RO, and SP packages and 1 day in the SLO package.