Accurate determination of succinimide degradation products using high fidelity trypsin digestion peptide map analysis

We report an efficient, high fidelity trypsin digestion method for peptide map analysis. This method minimizes artifacts caused by the sample preparation process, and we show its utility for the accurate determination of succinimide formation in a degraded monoclonal antibody product. A basic charge variant was detected by imaged capillary isoelectric focusing and was shown with reduced antigen binding and biological activity. Samples were reduced under denaturing conditions at pH 5.0, and digestion of the reduced protein with porcine trypsin was performed at pH 7.0 for 1 h. Following reversed phase high-performance liquid chromatography and online mass spectrometric analysis, succinimide formation was identified at Asp30 in the light chain. This result contrasts with the observation of only iso-Asp and Asp residues under conventional sample preparation conditions, which are therefore concluded to be artificially generated. The Asp30 residue is seen in the cocrystal structure model to participate in favorable charge interaction with an antigen molecule. Formation of succinimide and the resulting loss of negative charge are therefore hypothesized to be the degradation mechanism. After treatment of the degraded antibody sample to mildly alkaline pH conditions, we observed only Asp residue as the succinimide hydrolysis product and concurrent recovery of biological activity.     

Aquatic Training after Joint Immobilization in Rats Promotes Adaptations in Myotendinous Junctions

The myotendinous junction (MTJ) is the muscle-tendon interface and constitutes an integrated mechanical unit to force transmission. Joint immobilization promotes muscle atrophy via disuse, while physical exercise can be used as an adaptative stimulus. In this study, we aimed to investigate the components of the MTJ and their adaptations and the associated elements triggered with aquatic training after joint immobilization. Forty-four male Wistar rats were divided into sedentary (SD), aquatic training (AT), immobilization (IM), and immobilization/aquatic training (IMAT) groups. The samples were processed to measure fiber area, nuclear fractal dimension, MTJ nuclear density, identification of telocytes, sarcomeres, and MTJ perimeter length. In the AT group, the maintenance of ultrastructure and elements in the MTJ region were observed; the IM group presented muscle atrophy effects with reduced MTJ perimeter; the IMAT group demonstrated that aquatic training after joint immobilization promotes benefits in the muscle fiber area and fractal dimension, in the MTJ region shows longer sarcomeres and MTJ perimeter. We identified the presence of telocytes in the MTJ region in all experimental groups. We concluded that aquatic training is an effective rehabilitation method after joint immobilization due to reduced muscle atrophy and regeneration effects on MTJ in rats.     

Erratum: Development of composites based on recycled polyethylene/sugarcane bagasse fibers

When this article was first published in Polymer Composites, 35:768–774, 2014; DOI: 10.1002/pc.22720 , the lead authors name was incorrectly supplied to the Publisher. His correct name appears above. The authors apologize for any inconvenience this may have caused.     

Gamma radiation induced oxidation and tocopherols decrease in in-shell, peeled and blanched peanuts

In-shell, peeled and blanched peanut samples were characterized in relation to proximate composition and fatty acid profile. No difference was found in relation to its proximate composition. The three major fatty acids were palmitic acid, oleic acid, and linoleic acid. In order to investigate irradiation and storage effects, peanut samples were submitted to doses of 0.0, 5.0, 7.5 or 10.0 kGy, stored for six months at room temperature and monitored every three months. Peanuts responded differently to irradiation, particularly with regards to tocopherol contents, primary and secondary oxidation products and oil stability index. Induction periods and tocopherol contents were negatively correlated with irradiation doses and decreased moderately during storage. α-Tocopherol was the most gamma radiation sensitive and peeled samples were the most affected. A positive correlation was found among tocopherol contents and the induction period of the oils extracted from irradiated samples. Gamma radiation and storage time increased oxidation compounds production. If gamma radiation is considered an alternative for industrial scale peanut conservation, in-shell samples are the best feedstock. For the best of our knowledge this is the first article with such results; this way it may be helpful as basis for future studies on gamma radiation of in-shell crops.     

Extracorporeal hydroxyapatite-chamber for bone and biomaterial studies

Hydroxyapatite (HA) spherules and autologous bone (AB) with a central vascular pedicle were housed inside an HA-chamber to form the skeletal segment of specific shape. Experimental chambers were then inserted in a pocket between medial thigh muscles in 13 New Zealand male rabbits for 3 months. Three graft group were scheduled: (A) HA and AB without vascular pedicle, (B) HA with vascular pedicle, (C) HA and AB with vascular pedicle. At term, histology showed tissue and cellular degeneration in group A chambers. Due to spherules coalescence, fibrous tissue is formed in group B chambers. Group C chambers contained living osteocytes in the implanted bone, several newly formed vessels in soft tissue, bone and partial hydroxyapatite erosions. New bone was formed in apposition to both autologous bone and hydroxyapatite. Our study suggests that this experimental model could be used to grow adequately sized vascularized skeletal segments.     

Electronic structure of surface-supported bis(phthalocyaninato) terbium(III) single molecular magnets

The electronic structure of isolated bis(phthalocyaninato) terbium(III) molecules, a novel single-molecular-magnet (SMM), supported on the Cu(111) surface has been characterized by density functional theory and scanning tunneling spectroscopy. These studies reveal that the interaction with the metal surface preserves both the molecular structure and the large spin magnetic moment of the metal center. The 4f electron states are not perturbed by the adsorption while a strong molecular/metal interaction can induce the suppression of the minor spin contribution delocalized over the molecular ligands. The calculations show that the inherent spin magnetic moment of the molecule is only weakly affected by the interaction with the surface and suggest that the SMM character might be preserved.     

Polymer electrolytes for dye-sensitized solar cells prepared by photopolymerization of PEG-based oligomers

We report on the preparation and characterization of novel polymer electrolyte membranes for quasi-solid dye-sensitized solar cells. New methacrylic–acrylic gel-polymer electrolytes were prepared by photo-polymerization of mono/di-functional monomers. The crosslinked films were self standing, transparent and flexible. They were swelled by an iodine–iodide solution, obtaining a stable gel, where the polymeric network acts as a cage to retain the liquid, preventing its evaporation. Such a system combines the cohesive property of a solid with the high ionic conductivity of a liquid. The evaluation of the structural and physical-chemical characteristics of the polymer, combined with the electrical characterization of the membranes by means of the electrochemical impedance spectroscopy, allowed us to investigate the structure/property relationship of the material. The electric characterizations of the solar harvester based on the gel-polymer electrolyte showed a maximum photovoltaic conversion efficiency of 4.41%. Moreover, a significant improvement in the durability of the device was demonstrated with respect to the liquid electrolyte-based counterpart.     

Transgenic corn seed for recombinant protein production: relevant aspects on the aqueous extraction of native components

Plants have been considered one of the most promising expression systems for the production of recombinant proteins. Foreseen advantages include high productivity, optimal processing and assembly, and the non‐propagation of human or animal pathogens. A few successful examples of commercial proteins produced in plants have been reported in the literature, such us β‐glucoronidase, avidin, hirudin and aprotinin. Although the purification scheme is always a challenge in downstream processing development, the extraction is the key point since the presence of impurities deleterious to the process efficiency and operational‐life span of the equipment is determined at this step. This work reports the effect of pH and ionic strength in the extraction of proteins, phenolic compounds, lipids and sugars from transgenic corn seed. The phenolic compounds, lipids and reducing sugars were not significantly affected by changes in the ionic strength of the extracting solutions in the range 0–300 mM NaCl and pH 6.3. However, at high pH value (pH 10.0), high solubilization of proteins, phenolic compounds and lipids was achieved, whereas reducing sugars were not significantly extracted at this condition. This work is complementary to the studies reported in a previous paper and contributes to the development of recombinant protein recovery and purification process from transgenic corn. Copyright © 2004 Society of Chemical Industry     

Olfactometric approach for the evaluation of citizens' exposure to industrial emissions in the city of Terni, Italy

Since several years odour nuisance is a serious environmental concern in the city of Terni, whose citizens are repeatedly lamenting the presence of malodours. This paper describes the olfactometric approach adopted for assessing the odour impact on the city of Terni, caused by the co-presence of three important industrial poles: the steel industry pole, the chemical pole and a third industrial pole, comprising different activities for the treatment of wastewaters and solid waste. The combination of analyses by dynamic olfactometry and dispersion modelling allowed the evaluation of the citizens' exposure to the industrial odours, resulting in both the quantification of emissions and the assessment of their impact on the territory. The overall odour emissions were estimated to be equal to 218,000 ou_E s^− 1: 51% from the steel industry pole, 29% from the chemical pole and 20% from the other plants, respectively. The simulation of the emission dispersion shows that the odour impact relevant to all three studied industrial poles is considerable, actually investing almost the whole city of Terni. The study results also enabled the identification of the most problematic odour sources, which turned out to be the primary emissions from the furnace for the steel industry pole, Treofan and the wastewater treatment plant for the chemical pole and the municipal wastewater treatment plant and the pulper incinerator as far as the other plants are concerned.     

Non-thermal atmospheric gas plasma device for surface decontamination of shell eggs

A resistive barrier discharge (RBD) prototype able to generate gas plasma at atmospheric conditions was set up. The discharge was electrically characterized and the plasma glow was analysed by optical emission spectroscopy. The decontamination power of the device was assessed on samples of shell eggs experimentally inoculated with Salmonella Enteritidis and Salmonella Typhimurium (5.5–6.5 Log CFU/eggshell) and placed in the treatment chamber. Different decontamination times (10, 20, 30, 45, 60 and 90 min) and relative humidity values (RH) of the gas mixture in the chamber (i.e. 35% and 65%, at 25 °C) were considered. All samples were treated in the plasma after-glow chamber where the measured temperature was not much higher than the room temperature, minimizing the risk of egg quality alterations.