Philipp R. Stoessel

Reversible DNA encapsulation in silica to produce ROS-resistant and heat-resistant synthetic DNA 'fossils'

This protocol describes a method for encapsulating DNA into amorphous silica (glass) spheres, mimicking the protection of nucleic acids within ancient fossils. In this approach, DNA encapsulation is achieved after the ammonium functionalization of silica nanoparticles. Within the glass spheres, the nucleic acid molecules are hermetically sealed and protected from chemical attack, thereby withstanding high temperatures and aggressive radical oxygen species (ROS). The encapsulates can be used as inert taggants to trace chemical and biological entities. The present protocol is applicable to short double-stranded (ds) and single-stranded (ss) DNA fragments, genomic DNA and plasmids. The nucleic acids can be recovered from the glass spheres without harm by using fluoride-containing buffered oxide etch solutions. Special emphasis is placed in this protocol on the safe handling of these buffered hydrogen fluoride solutions. After dissolution of the spheres and subsequent purification, the nucleic acids can be analyzed by standard techniques (gel electrophoresis, quantitative PCR (qPCR) and sequencing). The protocol requires 6 d for completion with a total hands-on time of 4 h.

Labeling milk along its production chain with DNA encapsulated in silica

The capability of tracing a food product along its production chain is important to ensure food safety and product authenticity. For this purpose and as an application example, recently developed Silica Particles with Encapsulated DNA (SPED) were added to milk at concentrations ranging from 0.1 to 100 ppb (μg per kg milk). Thereby the milk, as well as the milk-derived products yoghurt and cheese, could be uniquely labeled with a DNA tag. Procedures for the extraction of the DNA tags from the food matrixes were elaborated and allowed identification and quantification of previously marked products by quantitative polymerase chain reaction (qPCR) with detection limits below 1 ppb of added particles. The applicability of synthetic as well as naturally occurring DNA sequences was shown. The usage of approved food additives as DNA carrier (silica = E551) and the low cost of the technology (<0.1 USD per ton of milk labeled with 10 ppb of SPED) display the technical applicability of this food labeling technology.

Physical mixtures of CeO2 and zeolites as regenerable indoor air purifiers: adsorption and temperature dependent oxidation of VOC

Removal of volatile organic compounds (VOC) and indoor air quality regulation through adsorbers required exchange or maintenance of active materials. In this work, we combine well known VOC adsorbers with oxidation catalysts as intimate particulate mixtures. We demonstrate how typical VOC can subsequently adsorb on such mixed material fixed beds (usually days to weeks; the common state of the system, adsorption phase) using small organic compounds (diethyl ether, triethylamine), monoterpenes such as linalool and limonene, and hexanoic acid. Occasional regeneration runs through heat up of the fixed bed results in simultaneous desorption and oxidation of the accumulated VOC, thus regenerating full adsorption capacity for a next adsorption phase. We investigated both small pore zeolites (H-ZSM-5) and larger pore zeolites (13X) and found a distinct interplay between the pore size and the type of VOC. Thermogravimetry coupled with mass spectroscopy was used to quantitatively study the effects of mixing composition and temperature on adsorber performance and regeneration. The here investigated bi-functional systems combine very low maintenance costs and materials requirement with low air flow and exchange costs, thus suggesting mixed (two-functional) bed adsorbers with catalytic function as sustainable alternatives to currently used single use systems based on granulated zeolites or activated carbon. In this work we show the ability of zeolite/cerium oxide physical mixtures to adsorb and capture different classes of VOC at room temperature and release them for oxidation at higher temperatures in a regenerative and sustainable process.

Incorporation of particulate bioactive glasses into a dental root canal sealer

Abstract Flame spray synthesis has opened the possibility to add additional elements to complex materials such as bioactive glasseswhile maintaining nanoparticulate properties. In this study, it was investigated whether a flamesprayed bismuth oxide doped nanometric 45S5 bioactive glass could be incorporated into a commercially available epoxy-resin root canal sealer, and how this compared to a conventional, pure 45S5 micrometric bioactive glass. Effects on radiopacity, microhardness, pH and mineral induction in phosphate buffered saline and simulated body fluid were studied. It was revealed that the radiopaque nanometric bismuth-containing 45S5 bioactive glass reduced radiopacity of the root canal sealer less than a conventional micrometric counterpart. In addition, pH induction and calcium phosphate precipitation were quicker with the nanometric compared to the micrometric material, whilst the micrometric glass displayed a higher alkaline capacity. Both materials apparently bound to the epoxy resin matrix, thus increasing its microhardness after polymerization reaction. Effects were dose-dependent. The investigated radiopaque bioactive glass containing bismuth oxide could be a valuable add-on for current root canal sealers.

PCR quantification of SiO2 particle uptake in cells in the ppb and ppm range via silica encapsulated DNA barcodes

There is a strong interest in studying the cellular uptake of silica nanoparticles, particularly at medically relevant concentrations (ppb-ppm range) to understand their toxicology. At present, uptake analysis at these exposure levels is impeded by the high silica background concentration. Here we describe the use of DNA encapsulated within silica particles as a tool to quantify silica nanoparticles in in vitro cell-uptake experiments at low concentrations (down to 10 fg cell(-1)).

Porous, Water-Resistant Multifilament Yarn Spun from Gelatin

Sustainability, renewability, and biodegradability of polymeric material constantly gain in importance. A plausible approach is the recycling of agricultural waste proteins such as keratin, wheat gluten, casein or gelatin. The latter is abundantly available from animal byproducts and may well serve as building block for novel polymeric products. In this work, a procedure for the dry-wet spinning of multifilament gelatin yarns was developed. The process stands out as precipitated gelatin from a ternary mixture (gelatin/solvent/nonsolvent) was spun into porous filaments. About 1000 filaments were twisted into 2-ply yarns with good tenacity (4.7 cN tex(-1)). The gelatin yarns, per se susceptible to water, were cross-linked by different polyfunctional epoxides and examined in terms of free lysyl amino groups and swelling degree in water. Ethylene glycol diglycidyl ether exhibited the highest cross-linking efficiency. Further post-treatments with gaseous formaldehyde and wool grease (lanolin) rendered the gelatin yarns water-resistant, allowing for multiple swelling cycles in water or in detergent solution. However, the swelling caused a decrease in filament porosity from ∼30% to just below 10%. To demonstrate the applicability of gelatin yarn in a consumer good, a gelatin glove with good thermal insulation capacity was fabricated.

Induced cyanogenesis from hydroxynitrile lyase and mandelonitrile on wheat with polylactic acid multilayer-coating produces self-defending seeds

Wheat is the predominant crop in the world and vermin infestation remains a serious issue regarding its storage and field cultivation, especially in developing countries. In addition to physical control methods, pesticides are often applied. However, their usage includes a number of concerns regarding environmental and ecological impacts. In the present work an alternative route to protect seeds against herbivore attack is suggested. The seeds were coated with substances capable of cyanogenesis. The precursors were initially in isolated compartments separated through biodegradable polylactide layers. The HCN formation only occurred upon contact of the cyanogenic precursor (mandelonitrile) with a suitable enzyme (hydroxynitrile lyase) and thus needed to be mechanically triggered. This concept is inspired by nature and is based on the protection strategy applied by higher plants, for example apple trees. Further tests showed that the ability for germination was preserved throughout the treatment. Finally, cyanogenesis was followed and quantified in both the liquid and the gas phase and provided HCN in sufficient concentrations to serve as a pest control.

Physical mixtures of CeO2 and zeolites as regenerable indoor air purifiers

Removal of volatile organic compounds (VOC) and indoor air quality regulation through adsorbers required exchange or maintenance of active materials. In this work, we combine well known VOC adsorbers with oxidation catalysts as intimate particulate mixtures. We demonstrate how typical VOC can subsequently adsorb on such mixed material fixed beds (usually days to weeks; the common state of the system, adsorption phase) using small organic compounds (diethyl ether, triethylamine), monoterpenes such as linalool and limonene, and hexanoic acid. Occasional regeneration runs through heat up of the fixed bed results in simultaneous desorption and oxidation of the accumulated VOC, thus regenerating full adsorption capacity for a next adsorption phase. We investigated both small pore zeolites (H-ZSM-5) and larger pore zeolites (13X) and found a distinct interplay between the pore size and the type of VOC. Thermogravimetry coupled with mass spectroscopy was used to quantitatively study the effects of mixing composition and temperature on adsorber performance and regeneration. The here investigated bi-functional systems combine very low maintenance costs and materials requirement with low air flow and exchange costs, thus suggesting mixed (two-functional) bed adsorbers with catalytic function as sustainable alternatives to currently used single use systems based on granulated zeolites or activated carbon. In this work we show the ability of zeolite/cerium oxide physical mixtures to adsorb and capture different classes of VOC at room temperature and release them for oxidation at higher temperatures in a regenerative and sustainable process.