Furan quantification in bread crust: development of a simple and sensitive method using headspace-trap GC-MS

To study reactivity in bread crust during the baking process in the pan, we followed furan mainly resulting from Maillard and caramelisation reactions in cereal products. Furan quantification is commonly performed with automatic HS-static GC-MS. However, we showed that the automatic HS-trap GC-MS method can improve the sensitivity of the furan quantification. Indeed, this method allowed the LOD to be decreased from 0.3 ng g^–1 with HS-static mode to 0.03 ng g^–1 with HS-trap mode under these conditions. After validation of this method for furan quantification in bread crust, a difference between the crust extracted from the bottom and from the sides of the bread was evident. The quantity of furan in the bottom crust was five times lower than in the side crust, revealing less reactivity on the bottom than on the sides of the bread during the baking process in the pan. Differences in water content may explain these variations in reactivity.     

Chemical and genetic wrappers for improved phage and RNA display

An Achilles heel inherent to all molecular display formats, background binding between target and display system introduces false positives into screens and selections. For example, the negatively charged surfaces of phage, mRNA, and ribosome display systems bind with unacceptably high nonspecificity to positively charged target molecules, which represent an estimated 35% of proteins in the human proteome. Here we report the first systematic attempt to understand why a broad class of molecular display selections fail, and then solve the underlying problem for both phage and RNA display. Firstly, a genetic strategy was used to introduce a short, charge-neutralizing peptide into the solvent-exposed, negatively charged phage coat. The modified phage (KO7(+)) reduced or eliminated nonspecific binding to the problematic high-pI proteins. In the second, chemical approach, nonspecific interactions were blocked by oligolysine wrappers in the cases of phage and total RNA. For phage display applications, the peptides Lys(n) (where n=16 to 24) emerged as optimal for wrapping the phage. Lys(8), however, provided effective wrappers for RNA binding in assays against the RNA binding protein HIV-1 Vif. The oligolysine peptides blocked nonspecific binding to allow successful selections, screens, and assays with five previously unworkable protein targets.     

Melectin: a novel antimicrobial peptide from the venom of the cleptoparasitic bee Melecta albifrons

A novel antimicrobial peptide designated melectin was isolated from the venom of the cleptoparasitic bee Melecta albifrons. Its primary sequence was established as H-Gly-Phe-Leu-Ser-Ile-Leu-Lys-Lys-Val-Leu-Pro-Lys-Val-Met-Ala-His-Met-Lys-NH(2) by Edman degradation and ESI-QTOF mass spectrometry. Synthetic melectin exhibited antimicrobial activity against both gram-positive and -negative bacteria and it degranulated rat peritoneal mast cells, but its hemolytic activity was low. The CD spectra of melectin measured in the presence of trifluoroethanol and sodium dodecyl sulfate showed a high content alpha-helices, which indicates that melectin can adopt an amphipathic alpha-helical secondary structure in an anisotropic environment such as the bacterial cell membrane. To envisage the role of the proline residue located in the middle of the peptide chain on biological activity and secondary structure, we prepared several melectin analogues in which the Pro11 residue was either replaced by other amino acid residues or was omitted. The results of biological testing suggest that a Pro kink in the alpha-helical structure of melectin plays an important role in selectivity for bacterial cells. In addition, a series of N- and C-terminal-shortened analogues was synthesized to examine which region of the peptide is related to antimicrobial activity.     

Distribution and characteristics of road traffic crashes in the Chaoyang District of Beijing, China

In many developing urban settings, economic growth and motorization are coupled with increasing rates of road traffic injuries (RTIs). By highlighting typical sites and circumstances at/in which car crashes occur, more specific targets for prevention can be identified. The study is based on police data for a 1-year period and covers the Chaoyang District, the biggest district of Beijing City. Focus is placed on crash patterns and their distribution by types of road and areas. Both fatal and non-fatal crashes are considered (n=754). In the main, the crashes occurred in relatively favorable driving circumstances (e.g., sunny weather, flat and straight roads, asphalted roads, and good traffic signals and road markings). They were also quite evenly distributed over time of day, day of week and season. Five crash patterns were highlighted, of which three were strongly associated with specific areas and four with specific types of road. The study supports the idea that type of road and RTI severity or pattern are closely related. It contributes to the development of context-relevant prevention measures aimed at reducing road crashes and minimizing their consequences and also supports safe planning of the road traffic environment.     

Soybean (Glycine max) Is Able to Absorb,Metabolize and Accumulate Fenbendazole in All Organs Including Beans

Although manure is an important source of minerals and organic compounds it represents a certain risk of spreading the veterinary drugs in the farmland and their permeation to human food. We tested the uptake of the anthelmintic drug fenbendazole (FBZ) by soybean, a common crop plant, from the soil and its biotransformation and accumulation in different soybean organs, including beans. Soybeans were cultivated in vitro or grown in a greenhouse in pots. FBZ was extensively metabolized in roots of in vitro seedlings, where sixteen metabolites were identified, and less in leaves, where only two metabolites were found. The soybeans in greenhouse absorbed FBZ by roots and translocated it to the leaves, pods, and beans. In roots, leaves, and pods two metabolites were identified. In beans, FBZ and one metabolite was found. FBZ exposure did not affect the plant fitness or yield, but reduced activities of some antioxidant enzymes and isoflavonoids content in the beans. In conclusion, manure or biosolids containing FBZ and its metabolites represent a significant risk of these pharmaceuticals entering food consumed by humans or animal feed. In addition, the presence of these drugs in plants can affect plant metabolism, including the production of isoflavonoids.     

Survey of air pollution in Cotonou, Benin--air monitoring and biomarkers

Exposure to genotoxic compounds present in ambient air has been studied in Cotonou, Benin, a city where two-stroke motorbikes are the major form of transportation and gasoline quality is poor. Personal monitoring and biomarkers were used to assess the exposure. Non-smoking taxi-moto drivers (city) and village residents were the study subjects. Benzene exposure was significantly higher in the city, as compared to the village (76.0+/-26.8 microg/m(3) versus 3.4+/-3.0, p=0.0004). Urinary excretion of benzene and S-phenylmercapturic acid (S-PMA) were also highest in subjects living in the city, whereas 1-hydroxypyrene was not different. The level of total polycyclic aromatic hydrocarbons (PAHs), associated with particles, ranged from 76.21 to 103.23 in Cotonou versus 1.55 ng/m(3) for the village. Determination of DNA damage in lymphocytes showed that subjects from the city had elevated number of lesions compared to subjects in the village in terms of bulky DNA adducts, 8-hydroxy-2'-deoxyguanosine and 5-methylcytosine, whereas DNA fragmentations analysed by alkaline gel electrophoresis was not different between the subjects. In conclusion, this study shows that air pollution is pronounced in Cotonou, Bénin and is associated with elevated levels of DNA damage in residents of the city compared to people living in a non-polluted rural village.     

Tie-Over Bolster Pressure Dressing Improves Outcomes of Skin Substitutes Xenografts on Athymic Mice

The efficacy of skin substitutes is established for the treatment of burn injuries, but its use is not limited to this condition. This technology has the potential to improve the treatment of various conditions by offering highly advanced and personalized treatments. In vivo studies are challenging but essential to move to clinical use in humans. Mice are the most widely used species in preclinical studies, but the main drawback of this model is the limited surface area of the graft in long-term transplantation studies caused by the displacement and the contraction of the graft. We improved the conventional surgical procedures by stabilizing the chamber covering the graft with intramuscular sutures and by adding a tie-over bolster dressing. The current study was therefore performed to compare outcomes of skin grafts between the conventional and optimized skin graft model. Human self-assembled skin substitutes (SASSs) were prepared and grafted to athymic mice either by the conventional method or by the new grafting method. Graft healing and complications were assessed using digital photographs on postoperative days 7, 14, and 21. Similar structure and organization were observed by histological staining. The new grafting method reduced medium and large displacement events by 1.26-fold and medium and large contraction events by 1.8-fold, leading to a 1.6-fold increase in graft surface area compared to skin substitutes grafted with the usual method. This innovation ensures better reproducibility and consistency of skin substitute transplants on mice.     

An E3‐14.7K Peptide that Promotes Microtubules‐Mediated Transport of Plasmid DNA Increases Polyplexes Transfection Efficiency

Chemical vectors as cationic polymers and cationic lipids are promising alternatives to viral vectors for gene therapy. Beside endosome escape and nuclear import, plasmid DNA (pDNA) migration in the cytosol toward the nuclear envelope is also regarded as a limiting step for efficient DNA transfection with non‐viral vectors. Here, the interaction between E3‐14.7K and FIP‐1 to favor migration of pDNA along microtubules is exploited. E3‐14.7K is an early protein of human adenoviruses that interacts via FIP‐1 (Fourteen.7K Interacting Protein 1) protein with the light‐chain components of the human microtubule motor protein dynein (TCTEL1). This peptide is conjugated with pDNA and mediates interaction of pDNA in vitro with isolated microtubules as well as with microtubules in cellulo. Videomicroscopy and tracking treatment of images clearly demonstrate that P79‐98/pDNA conjugate exhibits a linear transport with large amplitude along microtubules upon 2 h transfection with polyplexes whereas control pDNA conjugate exhibits small non‐directional movements in the cytoplasm. Remarkably, P79‐98/peGFP polyplexes enhance by a factor 2.5 (up to 76%) the number of transfected cells. The results demonstrate, for the first time, that the transfection efficiency of polyplexes can be drastically increased when the microtubules migration of pDNA is facilitated by a peptide allowing pDNA docking to TCTEL1. This is a real breakthrough in the non viral gene delivery field that opens hope to build artificial viruses.     

Membrane automata with priorities

In this paper the one-way P automata with priorities are introduced. Such automata are P systems where the membranes are only allowed to consume objects from parent membranes, under the given conditions. The result of computation of these systems is the set of multiset sequences consumed by skin membrane into the system. The rules associated in some order with each membrane cannot modify any objects, they can only move them through membrane. We show that P automata with priorities and two membranes can accept every recursively enumerated language.