Decellularised Human Umbilical Artery as a Vascular Graft Elicits Minimal Pro-Inflammatory Host Response Ex Vivo and In Vivo

Small diameter (<6 mm) vessel grafts still pose a challenge for scientists worldwide. Decellularised umbilical artery (dUA) remains promising as small diameter tissue engineered vascular graft (TEVG), yet their immunogenicity remains unknown. Herein, we evaluated the host immune responses, with a focus on the innate part, towards human dUA implantation in mice, and confirmed our findings in an ex vivo allogeneic human setup. Overall, we did not observe any differences in the number of circulating white blood cells nor the number of monocytes among three groups of mice (1) dUA patch; (2) Sham; and (3) Mock throughout the study (day −7 to 28). Likewise, we found no difference in systemic inflammatory and anti-inflammatory cytokine levels between groups. However, a massive local remodelling response with M2 macrophages were observed in the dUA at day 28, whereas M1 macrophages were less frequent. Moreover, human monocytes from allogeneic individuals were differentiated into macrophages and exposed to lyophilised dUA to maximize an eventual M1 response. Yet, dUA did not elicit any immediate M1 response as determined by the absence of CCR7 and CXCL10. Together this suggests that human dUA elicits a minimal pro-inflammatory response further supporting its use as a TEVG in an allogeneic setup.     

Sulfite Action in Glycolytic Inhibition: In Vivo Real‐Time Observation by Hyperpolarized 13C NMR Spectroscopy

Detecting the molecular targets of xenobiotic substances in vivo poses a considerable analytical challenge. Here, we describe the use of an NMR‐based tracer methodology for the instantaneous in vivo observation of sulfur(IV) action on cellular metabolism. Specifically, we find that glycolytic flux is directed towards sulfite adducts of dihydroxyacetone phosphate and pyruvate as off‐pathway intermediates that obstruct glycolytic flux. In particular, the pyruvate–sulfite association hinders the formation of downstream metabolites. The apparent in vivo association constant of pyruvate and sulfite agrees with the apparent inhibition constant of CO₂ formation, thus supporting the importance of pyruvate interception in disturbing central metabolism and inhibiting NAD regeneration.     

Real-time PCR quantification of the AM-toxin gene and HPLC qualification of toxigenic metabolites from Alternaria species from apples

Some Alternaria species are able to produce plant pathogenic as well as toxic metabolites. In both agriculture and the food industry it is important know if toxigenic Alternaria are present to rapidly employ the correct corrective actions. The purpose of this work was to establish a real-time PCR method, which can detect and quantify apple pathogenic and toxigenic Alternaria. An AM-toxin I primer set, which could recognize Alternaria DNA only, was designed by using primers complementary to the AM-toxin I gene. The method could detect small amounts of DNA (4 pg) and still obtain a large dynamic range (4 decades) without interference from apple material. Eight Alternaria isolates were analyzed for the presence of AM-toxin I gene and their production of secondary metabolites. Then analyses showed that all eight isolates contained the AM toxin gene and were able to produce the plant pathogenic tentoxin in addition to AM toxin I. The analyses also showed the production of tenuazonic acid, alternariols, Altenuene, altenusin and/or altertoxin I in pure culture. Analyses of inoculated apples showed that both the AM-toxin gene and alternariol monomethyl ether could be detected. Morphological analyses suggested that the eight Alternaria strains, though they all carried the AM toxin genes, probably belong to different but closely related un-described Alternaria taxa in the A. tenuissima species-group based on morphological and chemical differences.     

A screening experiment to identify factors causing rancidity during meat loaf production

A screening experiment consisting of 18 batches of meat loaves were made using a fractional factorial design with six factors including two production replicates i.e. testing of several factors in relatively few trials. The rancidity was detected by TBARS, volatile compounds by GC-MS and sensory analysis by a sensory panel. The results were presented graphically as formal statistical testing is limited. To do this the multivariate results from GC-MS and sensory analyses were made univariate by statistical tools. Pre-salting meat and storing it in frozen air, will result in rancid meat, and regardless pre-salting or not ascorbic acid will act as a pro-oxidant when added into already rancid meat and stored frozen. The three methods used to detect rancidity in meat loaves resulted sometimes in discrepancy in the results. Further research has to be done to verify if these findings were due to development of different oxidation products due to the combinations of factors or if these were outliers.     

Effects of prenatal exposure to diesel exhaust particles on postnatal development, behavior, genotoxicity and inflammation in mice

Background

Results from epidemiological studies indicate that particulate air pollution constitutes a hazard for human health. Recent studies suggest that diesel exhaust possesses endocrine activity and therefore may affect reproductive outcome. This study in mice aimed to investigate whether exposure to diesel exhaust particles (DEP; NIST 2975) would affect gestation, postnatal development, activity, learning and memory, and biomarkers of transplacental toxicity. Pregnant mice (C57BL/6; BomTac) were exposed to 19 mg/m³ DEP (~1·10⁶ particles/cm³; mass median diameter ? 240 nm) on gestational days 9–19, for 1 h/day.

Results

Gestational parameters were similar in control and diesel groups. Shortly after birth, body weights of DEP offspring were slightly lower than in controls. This difference increased during lactation, so by weaning the DEP exposed offspring weighed significantly less than the control progeny. Only slight effects of exposure were observed on cognitive function in female DEP offspring and on biomarkers of exposure to particles or genotoxic substances.

Conclusion

In utero exposure to DEP decreased weight gain during lactation. Cognitive function and levels of biomarkers of exposure to particles or to genotoxic substances were generally similar in exposed and control offspring. The particle size and chemical composition of the DEP and differences in exposure methods (fresh, whole exhaust versus aged, resuspended DEP) may play a significant role on the biological effects observed in this compared to other studies.     

Vitamin C,total phenolics and antioxidative activity in tip‐cut green beans (Phaseolus vulgaris) and swede rods (Brassica napus var. napobrassica) processed by methods used in catering

BACKGROUND: Retention of nutrients in vegetables during blanching/freezing, cooking and warm‐holding is crucial in the preparation of both standard and therapeutic diets. In the present study, conventional cooking in water, and cooking by pouch technology (boil‐in‐bag, sous vide) were compared in their ability to retain vitamin C, total phenolics and antioxidative activity (DPPH and FRAP) in industrially blanched/frozen tip‐cut green beans and swede rods. RESULTS: After conventional cooking, 50.4% total ascorbic acid, 76.7% total phenolics, 55.7% DPPH and 59.0% FRAP were recovered in the drained beans. After boil‐in‐bag cooking, significantly (P < 0.05) higher recoveries were obtained, i.e. 80.5% total ascorbic acid, 89.2% total phenolics, 94.8% DPPH and 92.9% FRAP. Recoveries after sous vide cooking were comparable to those of boil‐in‐bag cooking. By conventional cooking, 13.5–42.8% of the nutrients leaked into the cooking water; by sous vide about 10% leaked to the exuded liquid, while no leakage occurred by boil‐in‐bag cooking. Warm‐holding beans after cooking reduced recoveries in all components. Recoveries in swede rods were comparable but overall slightly lower. CONCLUSION: Industrially blanched/frozen vegetables should preferably be cooked by pouch technology, rather than conventional cooking in water. Including cooking water or exuded liquid into the final dish will increase the level of nutrients in a meal. Warm‐holding of vegetables after cooking should be avoided. Copyright © 2010 Society of Chemical Industry     

Reliability of industrial packaging for microsystems

Packaging concepts for silicon-based micromachined sensors exposed to harsh environments are explored. By exposing the sensors directly to the media and applying protection at the wafer level the packaging and assembly will be simplified as compared to conventional methods of fabrication.Protective coatings of amorphous silicon carbide and tantalum oxide are suitable candidates with etch rates below 0.1 Å/h in aqueous solutions with pH 11 at temperatures up to 140°C. Si-Ta-N films exhibit etch rates around 1 Å/h. Parylene C coatings did not etch but peeled off after extended exposure times at elevated temperatures. The best diamond-like carbon films we tested did not etch, but delaminated due to local penetration of the etchants.Several glue types were investigated for chip mounting of the sensors. Hard epoxies, such as Epotek H77, on the one hand exhibit high bond strength and least degradation and leakage, but on the other hand introduce large sensor output drift with temperature changes. Softening of the Epo-tek H77 was observed at 70°C.An industrially attractive thin-film anodic silicon-to-silicon wafer bonding process was developed. Glass layers are deposited at 20 nm/s (1.2 μm/min) by electron-beam evaporation and bond strengths in excess of 25 N/mm² are obtained for bonding temperatures higher than 300°C.Through-hole electrical feedthroughs with a minimum line width of 20μm and a density of 250 wires per cm were obtained by applying electro-depositable photo-resist. Hermetically sealed feedthroughs were obtained using glass frits, which withstand pressures of 4000 bar.     

Peroxidase,catalase and palmitoyl-CoA hydrolase activity in blanched carrot cubes after storage at −20°C

The effect of storage at ? 20°C was studied on carrot cubes which had been blanched at 50°, 60°, 70° and 80°C for 2, 5, 10 and 20 min. A sample of unblanched carrot cube was used as reference. Results from sensory methods were compared with the activity of the three enzymes, palmitoyl-CoA hydrolase, catalase and peroxidase. Good quality retention was obtained when palmitoyl-CoA hydrolase was no longer present, while activity of catalase (9%) and peroxidase (6%) remained.