Carlos A. Mora

Combining Phosphate and Bacteria Removal on Chemically Active Filter Membranes Allows Prolonged Storage of Drinking Water

A chemically active filtration membrane with incorporated lanthanum oxide nanoparticles enables the removal of bacteria and phosphate at the same time and thus provides a simple device for preparation of drinking water and subsequent safe storage without using any kind of disinfectants.

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.

In vitro activation of NAD‐dependent alcohol dehydrogenases by Nudix hydrolases is more widespread than assumed

In the Gram‐positive methylotroph Bacillus methanolicus, methanol oxidation is catalyzed by an NAD‐dependent methanol dehydrogenase (Mdh) that belongs to the type III alcohol dehydrogenase (Adh) family. It was previously shown that the in vitro activity of B. methanolicus Mdh is increased by the endogenous activator protein Act, a Nudix hydrolase. Here we show that this feature is not unique, but more widespread among type III Adhs in combination with Act or other Act‐like Nudix hydrolases. In addition, we studied the effect of site directed mutations in the predicted active site of Mdh and two other type III Adhs with regard to activity and activation by Act.

Hollow Carbon Nanobubbles: Synthesis, Chemical Functionalization, and Container-Type Behavior in Water.

Thin-walled, hollow carbon nanospheres with a hydrophobic interior and good water dispersability can be synthesized in two steps: First, metal nanoparticles, coated with a few layers of graphene-like carbon, are selectively modified on the outside with a covalently attached hydrophilic polymer. Second, the metal core is removed at elevated temperature treatment with acid, leaving a well-defined carbon-based hydrophobic cavity. Loading experiments with the dye rhodamine B and doxorubicin confirmed the filling and release of a cargo and adjustment of a dynamic equilibrium (cargo-loaded versus release). Rhodamine B preferably accumulates in the interior of the bubbles. Filled nanobubbles allowed constant dye release into pure water. Studies of the concentration-dependent loading and release show an unusual hysteresis.

DNA protection against ultraviolet irradiation by encapsulation in a multilayered SiO2/TiO2 assembly

DNA is protected against UV-induced damage by encapsulation in a core-shell-shell particulate construct. The DNA is hermetically sealed in SiO2 particles coated with TiO2. The TiO2 coating acts as a physical sunscreen and prevents high energy photons from damaging the nucleic acids. DNA can be recovered unharmed from the protection system with fluoride comprising buffers, and then directly analyzed using biochemical standard techniques (quantitative PCR, gel electrophoresis and Sanger sequencing). The coatings increase the DNA UV resistance by 42 times, which is equivalent to the increase in UV resistance obtained by bacteria during sporulation. The attenuation coefficient of the 20 nm titania layer is 1.8 106 cm-1 at 254 nm UV irradiation and optical attenuation is largely attributed to light scattering on the titania surface.

Detecting and Number Counting of Single Engineered Nanoparticles by Digital Particle Polymerase Chain Reaction

The concentrations of nanoparticles present in colloidal dispersions are usually measured and given in mass concentration (e.g. mg/mL), and number concentrations can only be obtained by making assumptions about nanoparticle size and morphology. Additionally traditional nanoparticle concentration measures are not very sensitive, and only the presence/absence of millions/billions of particles occurring together can be obtained. Here, we describe a method, which not only intrinsically results in number concentrations, but is also sensitive enough to count individual nanoparticles, one by one. To make this possible, the sensitivity of the polymerase chain reaction (PCR) was combined with a binary (=0/1, yes/no) measurement arrangement, binomial statistics and DNA comprising monodisperse silica nanoparticles. With this method, individual tagged particles in the range of 60-250 nm could be detected and counted in drinking water in absolute number, utilizing a standard qPCR device within 1.5 h of measurement time. For comparison, the method was validated with single particle inductively coupled plasma mass spectrometry (sp-ICPMS).

Silica particles with encapsulated DNA as trophic tracers

Ecological networks such as food webs are extremely complex and can provide important information about the robustness and productivity of an ecosystem. In most cases, it is not feasible to observe trophic interactions between predators and prey directly and with the available methods, it is difficult to quantify the connections between them. Here, we show that submicron‐sized silica particles (100–150 nm) with encapsulated DNA (SPED) enable accurate food and organism labelling and quantification of specific animal‐to‐animal transfer over more than one trophic level. We found that SPED were readily transferable and quantifiable from the bottom to the top of a two‐level food chain of arthropods. SPED were taken up in the gut system and remained persistent in an animal over several days. When uniquely labelled SPED were applied at predefined ratios, we found that information about their relative abundance was reliably conserved after trophic level transfer and over time. SPED were also applied to investigate the flower preference of fly pollinators and proved to be a fast and accurate analysis method. SPED combine attributes of DNA barcoding and stable isotope analysis such as unique labelling, quantification via real‐time PCR and exact backtracking to the tracer source. This improves and simplifies the analysis and monitoring of ecological networks.

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.

Application of the Prunus spp. Cyanide Seed Defense System onto Wheat: Reduced Insect Feeding and Field Growth Tests

Many crops are ill-protected against insect pests during storage. To protect cereal grains from herbivores during storage, pesticides are often applied. While pesticides have an undoubtable functionality, increasing concerns are arising about their application. In the present study, we investigated a bioinspired cyanogenic grain coating with amygdalin as cyanogenic precursor mimicking the feeding-triggered release of hydrogen cyanide (HCN) found for example in bitter almonds. The multilayer coating consisted of biodegradable polylactic acid with individual layers containing amygdalin or β-glucosidase which is capable of degrading amygdalin to HCN. This reaction occurred only when the layers were ruptured, e.g., by a herbivore attack. Upon feeding coated cyanogenic wheat grains to Tenebrio molitor (mealworm beetle), Rhizopertha dominica (lesser grain borer), and Plodia interpunctella (Indianmeal moth), their reproduction as well as consumption rate were significantly reduced, whereas germination ability increased compared to noncoated grains. In field experiments, we observed an initial growth delay compared to uncoated grains which became negligible at later growth stages. The here shown strategy to artificially apply a naturally occurring defense mechanisms could be expanded to other crops than wheat and has the potential to replace certain pesticides with the benefit of complete biodegradability and increased safety during storage.

Nondestructive in-line sub-picomolar detection of magnetic nanoparticles in flowing complex fluids

Over the last decades, the use of magnetic nanoparticles in research and commercial applications has increased dramatically. However, direct detection of trace quantities remains a challenge in terms of equipment cost, operating conditions and data acquisition times, especially in flowing conditions within complex media. Here we present the in-line, non-destructive detection of magnetic nanoparticles using high performance atomic magnetometers at ambient conditions in flowing media. We achieve sub-picomolar sensitivities measuring ~30 nm ferromagnetic iron and cobalt nanoparticles that are suitable for biomedical and industrial applications, under flowing conditions in water and whole blood. Additionally, we demonstrate real-time surveillance of the magnetic separation of nanoparticles from water and whole blood. Overall our system has the merit of in-line direct measurement of trace quantities of ferromagnetic nanoparticles with so far unreached sensitivities and could be applied in the biomedical field (diagnostics and therapeutics) but also in the industrial sector.