Evaluation of a Novel Thiol–Norbornene-Functionalized Gelatin Hydrogel for Bioprinting of Mesenchymal Stem Cells

Introduction: Three-dimensional bioprinting can be considered as an advancement of the classical tissue engineering concept. For bioprinting, cells have to be dispersed in hydrogels. Recently, a novel semi-synthetic thiolene hydrogel system based on norbornene-functionalized gelatin (GelNB) and thiolated gelatin (GelS) was described that resulted in the photoclick hydrogel GelNB/GelS. In this study, we evaluated the printability and biocompatibility of this hydrogel system towards adipose-tissue-derived mesenchymal stem cells (ASCs). Methods: GelNB/GelS was synthesized with three different crosslinking densities (low, medium and high), resulting in different mechanical properties with moduli of elasticity between 206 Pa and 1383 Pa. These hydrogels were tested for their biocompatibility towards ASCs in terms of their viability, proliferation and differentiation. The extrusion-based bioprinting of ASCs in GelNB/GelS-high was performed to manufacture three-dimensional cubic constructs. Results: All three hydrogels supported the viability, proliferation and chondrogenic differentiation of ASCs to a similar extent. The adipogenic differentiation of ASCs was better supported by the softer hydrogel (GelNB/GelS-low), whereas the osteogenic differentiation was more pronounced in the harder hydrogel (GelNB/GelS-high), indicating that the differentiation fate of ASCs can be influenced via the adaption of the mechanical properties of the GelNB/GelS system. After the ex vivo chondrogenic differentiation and subcutaneous implantation of the bioprinted construct into immunocompromised mice, the production of negatively charged sulfated proteoglycans could be observed with only minimal inflammatory signs in the implanted material. Conclusions: Our results indicate that the GelNB/GelS hydrogels are very well suited for the bioprinting of ASCs and may represent attractive hydrogels for subsequent in vivo tissue engineering applications.     

Determination of Glyphosate and Its Metabolite AMPA (Aminomethylphosphonic Acid) in Cereals After Derivatization by Isotope Dilution and UPLC-MS/MS”

A straightforward analytical method has been developed for the determination of glyphosate and aminomethylphosphonic acid (AMPA), its major metabolite in cereals. This method entails a rapid extraction and derivatization with 9-fluorenylmethyl chloroformate followed by separation with a conventional reversed-phase rapid chromatography used in daily routine analysis and detection by electrospray ionization tandem mass spectrometry. To overcome matrix effects and compensate for any analyte losses during sample treatment, an isotopic dilution approach was used. Since 2010, the monitoring of cereals for the widely used herbicide glyphosate is obligatory to all European Union (EU)-member states, laid down in Commission Regulation (EC) No. 1213/2008. Hence, there is definitively a need for a reliable and easy-to-handle analytical method for monitoring of this compound. The proposed method can be run without having to make time-consuming changes on the equipment used for daily routine analysis. The analytical performance of the method was evaluated according to SANCO/10684/2009 criteria and demonstrated that this method is rugged and cost-effective, therefore suitable for monitoring purposes as well as legislative enforcements within the EU. The apparent recoveries of both analytes were between 97% and 113% with relative standard deviations less than 20%. The limits of quantification of glyphosate and AMPA were 0.02 mg/kg in cereal matrices. Finally, the accuracy of the method was assessed by analyzing a proficiency test material which was available from a previous round (EUPT-C4).     

Adaptation of Oxidative Phosphorylation Machinery Compensates for Hepatic Lipotoxicity in Early Stages of MAFLD

Alterations in mitochondrial function are an important control variable in the progression of metabolic dysfunction-associated fatty liver disease (MAFLD), while also noted by increased de novo lipogenesis (DNL) and hepatic insulin resistance. We hypothesized that the organization and function of a mitochondrial electron transport chain (ETC) in this pathologic condition is a consequence of shifted substrate availability. We addressed this question using a transgenic mouse model with increased hepatic insulin resistance and DNL due to constitutively active human SREBP-1c. The abundance of ETC complex subunits and components of key metabolic pathways are regulated in the liver of these animals. Further omics approaches combined with functional assays in isolated liver mitochondria and primary hepatocytes revealed that the SREBP-1c-forced fatty liver induced a substrate limitation for oxidative phosphorylation, inducing enhanced complex II activity. The observed increased expression of mitochondrial genes may have indicated a counteraction. In conclusion, a shift of available substrates directed toward activated DNL results in increased electron flows, mainly through complex II, to compensate for the increased energy demand of the cell. The reorganization of key compounds in energy metabolism observed in the SREBP-1c animal model might explain the initial increase in mitochondrial function observed in the early stages of human MAFLD.     

Comparison of the performance of masterbatch and liquid color concentrates for mass coloration of polypropylene

The properties of polypropylene (PP) mixed with masterbatch (MB) and liquid color concentrates (LCC), respectively, were compared by preparing samples of PP with MB and PP with LCC and neat PP as a reference material using 1–4 extrusion cycles. Two colors were examined, i.e., a white color consisting of pigment white 6, and a green color consisting of pigment white 6, pigment blue 15:3, and pigment green 7. The color difference between PP prepared with MB and LCC was determined and the mechanical, rheological, and crystalline properties of PP prepared with MB and LCC were found to be generic. The color of PP obtained from MB and LCC were comparable. Further, it was found that the tensile strength, the viscosity, and the crystal structure obtained were similar when using LCC instead of MB. The viscosity of the treated PP generally decreased with increasing extruder retention time due to thermal degradation. The tensile strength and the crystal structure were independent of increased extruder retention time. © 2015 Wiley Periodicals, Inc. Col Res Appl, 41, 484–492, 2016     

Effect of maturation conditions on sensory and antioxidant properties of old Serbian plum brandies

?ljivovica and Prepe?enica are traditional Serbian distillated beverages made from varieties of plum, of the genus Prunus. The ageing in wooden casks not only improves the sensory characteristics, but also changes the content of total polyphenols and contributes to the other important health properties, such as the increase of their antioxidant capacity. In this study, home-made brandy (?ljivovica and Prepe?enica) samples were used, which matured in casks during the time period from 10 to 47?years. The aim of this investigation was to examine the influence of the ageing parameters (time period and type of wooden casks) and raw material (plum cultivars) on the total polyphenol content, amount of certain phenol compounds, sensory properties, colour and antioxidant properties in matured brandies. The individual phenol components were determinated using HPLC system equipped with diode array detector, while the total phenol content and antioxidant activity was estimated using spectrophotometric methods (Folin?CCiocalteu, DPPH, FRAP and TEAC). Total polyphenol content (TPC) of aged Prepe?enica samples was ranged from 230.26 to 890.26?mg/L gallic acid equivalents (GAE mg/L), whereas the TPC of ?ljivovica sample was 110.38?mg/L. Antioxidant capacity of analysed Prepe?enica samples and ?ljivovica sample was in strong correlation with TPC. Total sensory score of all samples was very high and varied between 18.2 and 19.8. The obtained results have shown that the benefit of longer maturation on sensory characteristics is well recognized, but the improvement has some limitation.     

Diamine oxidase supplementation improves symptoms in patients with histamine intolerance

Food Science and Biotechnology - Histamine intolerance (HIT) is thought to be caused by a disproportionate amount of histamine in the body. The enzyme diamine oxidase (DAO) is considered for the...     

The use and nature of present-focused interventions in cognitive and behavioral therapies for depression.

To improve cognitive and behavioral therapies (CBT) for depression, several approaches recommend an increased focus on the occurrence of problems as they occur in the therapeutic relationship or in relation to the live therapy process, referred to as present-focused. A lingering question has been the degree to which CBT therapists already engage in present-focused work. This study utilized sessions from recent trials of CBT for depression and, in Phase I, raters identified present-focused interventions on a turn-by-turn basis. Phase II raters used a qualitative analysis to determine categories of present-focused interventions. Results indicated that therapists rarely focused on the therapeutic relationship; when they did, it was often transient and lacking in the elaborations suggested by newer approaches. Therapists more often performed therapy process and emotion focused interventions, but these also tended to lack elaboration. (PsycINFO Database Record (c) 2010 APA, all rights reserved)