Nanomotors: new strategies of (bio)sensing based on motion

Nanomotors are nanoscale devices capable of converting energy into movement and forces. Among them, self-propelled nanomotors offer considerable promise for developing new and novel bioanalytical and biosensing strategies based on the direct isolation of target biomolecules or changes in their movement in the presence of target analytes. The mainachievements of this project consists on the development of receptor-functionalized nanomotors that offer direct and rapid target detection, isolation and transport from raw biological samples without preparatory and washing steps. For example, microtube engines functionalized with aptamer, antibody, lectin and enzymes receptors were used for the direct isolation of analytes of biomedical interest, including proteins and whole cells, among others. A target protein was also isolated from a complex sample by using an antigen-functionalized microengine navigating into the reservoirs of a lab-on-a-chip device. The new nanomotorbased target biomarkers detection strategy not only offers highly sensitive, rapid, simple and low cost alternative for the isolation and transport of target molecules, but also represents a new dimension of analytical information based on motion. The recognition events can be easily visualized by optical microscope (without any sophisticated analytical instrument) to reveal the target presence and concentration. The use of artificial nanomachines has shown not only to be useful for (bio)recognition and (bio)transport but also for detection of environmental contamination and remediation. In this context, micromotors modified with superhydrophobic layer demonstrated that effectively interacted, captured, transported and removed oil droplets from oil contaminated samples. Finally, a unique micromotor-based strategy for water-quality testing, that mimics live-fish water-quality testing, based on changes in the propulsion behavior of artificial biocatalytic microswimmers in the presence of aquatic pollutants was also developed. The attractive features of the new micromachine-based target isolation and signal transduction protocols developed in this project offer numerous potential applications in biomedical diagnostics, environmental monitoring, and forensic analysis.

Characterization of olive oil by carbon isotope analysis of individual fatty acids: Implications for authentication

The fatty acids of olive oils of distinct quality grade from the most important European Union (EU) producer countries were chemically and isotopically characterized. The analytical approach utilized combined capillary column gas chromatography-mass spectrometry (GC/MS) and the novel technique of compound-specific isotope analysis (CSIA) through gas chromatography coupled to a stable isotope ratio mass spectrometer (IRMS) via a combustion (C) interface (GC/C/IRMS). This approach provides further insights into the control of the purity and geographical origin of oils sold as cold-pressed extra virgin olive oil with certified origin appellation. The results indicate that substantial enrichment in heavy carbon isotope (C-13) of the bulk oil and of individual fatty acids are related to (1) a thermally induced degradation due to deodorization or steam washing of the olive oils and (2) the potential blend with refined olive oil or other vegetable oils. The interpretation of the data is based on principal component analysis of the fatty acids concentrations and isotopic data (delta(13)C(oil), delta(13)C(16:0), delta(13)C(18:1)) and on the delta(13)C(16:0) vs delta(13)C(18:1) covariations. The differences in the delta(13)C values of palmitic and oleic acids are discussed in terms of biosynthesis of these acids in the plant tissue and admixture of distinct oils.

The Long-Run Impact of Corn-Based Ethanol on the Grain, Oilseed, and Livestock Sectors: A Preliminary Assessment, November 2006

The ongoing growth of corn-based ethanol production raises some fundamental questions about what impact continued growth will have on U.S. and world agriculture. Estimates of the long-run potential for ethanol production can be made by calculating the corn price at which the incentive to expand ethanol production disappears. Under current ethanol tax policy, if the prices of crude oil, natural gas, and distillers grains stay at current levels, then the break-even corn price is $4.05 per bushel. A multi-commodity, multi country system of integrated commodity models is used to estimate the impacts if we ever get to $4.05 corn. At this price, corn-based ethanol production would reach 31.5 billion gallons per year, or about 20% of projected U.S. fuel consumption in 2015. Supporting this level of production would require 95.6 million acres of corn to be planted. Total corn production would be approximately 15.6 billion bushels, compared to 11.0 billion bushels today. Most of the additional corn acres come from reduced soybean acreage. Wheat markets would adjust to fulfill increased demand for feed wheat. Corn exports and production of pork and poultry would all be reduced in response to higher corn prices and increased utilization of corn by ethanol plants. These results should not be viewed as a prediction of what will eventually materialize. Rather, they indicate a logical end point to the current incentives to invest in corn-based ethanol plants.

Development of bioreporter assays for the detection of bioavailability of long-chain alkanes based on the marine bacterium Alcanivorax borkumensis strain SK2.

Long-chain alkanes are a major component of crude oil and therefore potentially good indicators of hydrocarbon spills. Here we present a set of new bacterial bioreporters and assays that allow to detect long-chain alkanes. These reporters are based on the regulatory protein AlkS and the alkB1 promoter from Alcanivorax borkumensis SK2, a widespread alkane degrader in marine habitats. Escherichia coli cells with the reporter construct reacted strongly to octane in short-term (6 h) aqueous suspension assays but very slightly only to tetradecane, in line with what is expected from its low water solubility. In contrast, long-term assays (up to 5 days) with A. borkumensis bioreporters showed strong induction with tetradecane and crude oil. Gel-immobilized A. borkumensis reporter cells were used to demonstrate tetradecane and crude oil bioavailability at a distance from a source. Alcanivorax borkumensis bioreporters induced fivefold more rapid and more strongly when allowed physical contact with the oil phase in standing flask assays, suggesting a major contribution of adhered cells to the overall reporter signal. Using the flask assays we further demonstrated the effect of oleophilic nutrients and biosurfactants on oil availability and degradation by A. borkumensis. The fluorescence signal from flask assays could easily be captured with a normal digital camera, making such tests feasible to be carried out on, e.g. marine oil responder vessels in case of oil accidents.

CO2, CH4 and N2O fluxes in an Ultisol treated with sewage sludge and cultivated with castor bean

Organic residue application into soil alter the emission of gases to atmosphere and CO2, CH4, N2O may contribute to increase the greenhouse effect. This experiment was carried out in a restoration area on a dystrophic Ultisol (PVAd) to quantify greenhouse gas (GHG) emissions from soil under castor bean cultivation, treated with sewage sludge (SS) or mineral fertilizer. The following treatments were tested: control without N; FertMin = mineral fertilizer; SS5 = 5 t ha-1 SS (37.5 kg ha-1 N); SS10 = 10 t ha-1 SS (75 kg ha-1 N); and SS20 = 20 t ha-1 SS (150 kg ha-1 N). The amount of sludge was based on the recommended rate of N for castor bean (75 kg ha-1), the N level of SS and the mineralization fraction of N from SS. Soil gas emission was measured for 21 days. Sewage sludge and mineral fertilizers altered the CO2, CH4 and N2O fluxes. Soil moisture had no effect on GHG emissions and the gas fluxes was statistically equivalent after the application of FertMin and of 5 t ha-1 SS. The application of the entire crop N requirement in the form of SS practically doubled the Global Warming Potential (GWP) and the C equivalent emissions in comparison with FertMin treatments.

Diagnosis and recommendation integrated system (DRIS) of soybean seed oil content

The Diagnosis and Recommendation Integrated System (DRIS) can improve interpretations of leaf analysis to determine the nutrient status. Diagnoses by this method require DRIS norms, which are however not known for oil content of soybean seeds. The aims of this study were to establish and test the DRIS method for oil content of soybean seed (maturity group II cultivars). Soybean leaves (207 samples) in the full flowering stage were analyzed for macro and micro-nutrients, and the DRIS was applied to assess the relationship between nutrient ratios and the seed oil content. Samples from experimental and farm field sites of the southernmost Brazilian state Rio Grande do Sul (28° - 29° southern latitude; 52° -53° western longitude) were assessed in two growing seasons (2007/2008 and 2008/2009). The DRIS norms related to seed oil content differed between the studied years. A unique DRIS norm was established for seed oil content higher than 18.68 % based on data of the 2007/2008 growing season. Higher DRIS indices of B, Ca, Mg and S were associated with a higher oil content, while the opposite was found for K, N and P. The DRIS can be used to evaluate the leaf nutrient status of soybean to improve the seed oil content of the crop.

Foam pore size is a critical interface parameter of suction-based wound healing devices.

BACKGROUND: Suction-based wound healing devices with open-pore foam interfaces are widely used to treat complex tissue defects. The impact of changes in physicochemical parameters of the wound interfaces has not been investigated. METHODS: Full-thickness wounds in diabetic mice were treated with occlusive dressing or a suction device with a polyurethane foam interface varying in mean pore size diameter. Wound surface deformation on day 2 was measured on fixed tissues. Histologic cross-sections were analyzed for granulation tissue thickness (hematoxylin and eosin), myofibroblast density (α-smooth muscle actin), blood vessel density (platelet endothelial cell adhesion molecule-1), and cell proliferation (Ki67) on day 7. RESULTS: Polyurethane foam-induced wound surface deformation increased with polyurethane foam pore diameter: 15 percent (small pore size), 60 percent (medium pore size), and 150 percent (large pore size). The extent of wound strain correlated with granulation tissue thickness that increased 1.7-fold in small pore size foam-treated wounds, 2.5-fold in medium pore size foam-treated wounds, and 4.9-fold in large pore size foam-treated wounds (p < 0.05) compared with wounds treated with an occlusive dressing. All polyurethane foams increased the number of myofibroblasts over occlusive dressing, with maximal presence in large pore size foam-treated wounds compared with all other groups (p < 0.05). CONCLUSIONS: The pore size of the interface material of suction devices has a significant impact on the wound healing response. Larger pores increased wound surface strain, tissue growth, and transformation of contractile cells. Modification of the pore size is a powerful approach for meeting biological needs of specific wounds.

First day of an oil spill on the open sea: early mass transfers of hydrocarbons to air and water.

During the first hours after release of petroleum at sea, crude oil hydrocarbons partition rapidly into air and water. However, limited information is available about very early evaporation and dissolution processes. We report on the composition of the oil slick during the first day after a permitted, unrestrained 4.3 m(3) oil release conducted on the North Sea. Rapid mass transfers of volatile and soluble hydrocarbons were observed, with >50% of ≤C17 hydrocarbons disappearing within 25 h from this oil slick of <10 km(2) area and <10 μm thickness. For oil sheen, >50% losses of ≤C16 hydrocarbons were observed after 1 h. We developed a mass transfer model to describe the evolution of oil slick chemical composition and water column hydrocarbon concentrations. The model was parametrized based on environmental conditions and hydrocarbon partitioning properties estimated from comprehensive two-dimensional gas chromatography (GC×GC) retention data. The model correctly predicted the observed fractionation of petroleum hydrocarbons in the oil slick resulting from evaporation and dissolution. This is the first report on the broad-spectrum compositional changes in oil during the first day of a spill at the sea surface. Expected outcomes under other environmental conditions are discussed, as well as comparisons to other models.

Development of Bio-Based Polymers for Use in Asphalt, TR-639, 2014

Asphalt binder is typically modified with poly type (styrene-butadiene-styrene or SBS) polymers to improve its rheological properties and performance grade. The elastic and principal component of SBS polymers is butadiene. For the last decade, butadiene prices have fluctuated and significantly increased, leading state highway agencies to search for economically viable alternatives to butadiene based materials. This project reports the recent advances in polymerization techniques that have enabled the synthesis of elastomeric, thermoplastic, block-copolymers (BCPs) comprised of styrene and soybean oil, where the “B” block in SBS polymers is replaced with polymerized triglycerides derived from soybean oil. These new breeds of biopolymers have elastomeric properties comparable to well-established butadiene-based styrenic BCPs. In this report, two types of biopolymer formulations are evaluated for their ability to modify asphalt binder. Laboratory blends of asphalt modified with the biopolymers are tested for their rheological properties and performance grade. Blends of asphalt modified with the biopolymers are compared to blends of asphalt modified with two commonly used commercial polymers. The viscoelastic properties of the blends show that biopolymers improve the performance grade of the asphalt to a similar and even greater extent as the commercial SBS polymers. Results shown in this report indicate there is an excellent potential for the future of these biopolymers as economically and environmentally favorable alternatives to their petrochemically-derived analogs.

A new green fluorescent protein-based bacterial biosensor for analysing phenanthrene fluxes.

The polycyclic aromatic hydrocarbon (PAH)-degrading strain Burkholderia sp. RP007 served as host strain for the design of a bacterial biosensor for the detection of phenanthrene. RP007 was transformed with a reporter plasmid containing a transcriptional fusion between the phnS putative promoter/operator region and the gene encoding the enhanced green fluorescent protein (GFP). The resulting bacterial biosensor--Burkholderia sp. strain RP037--produced significant amounts of GFP after batch incubation in the presence of phenanthrene crystals. Co-incubation with acetate did not disturb the phenanthrene-specific response but resulted in a homogenously responding population of cells. Active metabolism was required for induction with phenanthrene. The magnitude of GFP induction was influenced by physical parameters affecting the phenanthrene flux to the cells, such as the contact surface area between solid phenanthrene and the aqueous phase, addition of surfactant, and slow phenanthrene release from Model Polymer Release System beads or from a water-immiscible oil. These results strongly suggest that the bacterial biosensor can sense different phenanthrene fluxes while maintaining phenanthrene metabolism, thus acting as a genuine sensor for phenanthrene bioavailability. A relationship between GFP production and phenanthrene mass transfer is proposed.

Antigen sparing with adjuvanted inactivated polio vaccine based on Sabin strains.

Six different adjuvants, each in combination with inactivated polio vaccine (IPV) produced with attenuated Sabin strains (sIPV), were evaluated for their ability to enhance virus neutralizing antibody titres (VNTs) in the rat potency model. The increase of VNTs was on average 3-, 15-, 24-fold with adjuvants after one immunization (serotypes 1, 2, and 3, respectively). Also after a boost immunization the VNTs of adjuvanted sIPV were on average another 7-20-27 times higher than after two inoculations of sIPV without adjuvant. The results indicate that it is feasible to increase the potency of inactivated polio vaccines by using adjuvants.

Development of Non-Petroleum Based Binders for Use in Flexible Pavements, TR-594, 2010

Most bituminous adhesives or binders that are used for pavement materials are derived primarily from fossil fuels. With petroleum oil reserves becoming depleted and the drive to establish a bio-based economy, there is a push to produce binders from alternative sources, particularly from biorenewable resources. However, until now, no research has studied the applicability of utilizing bio-oils as a bitumen replacement (100% replacement) in the pavement industry. The main objective of this research was to test various properties of bio-oils in order to determine the applicability of using bio-oils as binders in the pavement industry. The overall conclusions about the applicability of using bio-oils as bio-binders in the pavement industry can be summarized as follows: 1. Bio-oils cannot be used as bio-binders/pavement materials without any heat pre-treatment/upgrading procedure. 2. Current testing standards and specifications, especially Superpave procedures, should be modified to comply with the properties of bio-binders. 3. The temperature range of the viscous behavior for bio-oils may be lower than that of bitumen binders by about 30°–40° C. 4. The rheological properties of the unmodified bio-binders vary in comparison to bitumen binders, but the rheological properties of these modified bio-binders change significantly upon adding polymer modifiers. 5. The high-temperature performance grade for the developed bio-binders may not vary significantly from that of the bitumen binders, but the low-temperature performance grade may vary significantly

Development of Non-Petroleum-Based Binders for Use in Flexible Pavements – Phase II, TR-650, 2015

Bio-binders can be utilized as asphalt modifiers, extenders, and replacements for conventional asphalt in bituminous binders. From the rheology results of Phase I of this project, it was found that the bio-binders tested had good performance, similar to conventional asphalt, except at low temperatures. Phase II of this project addresses this shortcoming and evaluates the Superpave performance of laboratory mixes produced with the enhanced bio-binders. The main objective of this research was to develop a bio-binder capable of replacing conventional asphalt in flexible pavements by incorporating ground tire rubber (GTR) into bio-oil derived from fast pyrolysis of agriculture and forestry residues. The chemical compatibility of the new bio-binder with GTR was assessed, and the low-temperature performance of the bio-binders was enhanced by the use of GTR. The newly developed binder, which consisted of 80 percent conventional binder and 20 percent rubber-modified bio-oil (85 percent bio-oil with 15 percent GTR), was used to produce mixes at two different air void contents, 4 and 7 percent. The laboratory performance test results showed that the performance of the newly developed bio-binder mixes is as good as or better than conventional asphalt mixes for fatigue cracking, rutting resistance, moisture sensitivity, and low-temperature cracking. These results need to be validated in field projects in order to demonstrate adequate performance for this innovative and sustainable technology for flexible pavements.

Unlike PAHs from Exxon Valdez crude oil, PAHs from Gulf of Alaska coals are not readily bioavailable.

In the wake of the 1989 Exxon Valdez oil spill, spatially and temporally spill-correlated biological effects consistent with polycyclic aromatic hydrocarbon (PAH) exposure were observed. Some works have proposed that confounding sources from local source rocks, prominently coals, are the provenance of the PAHs. Representative coal deposits along the southeast Alaskan coast (Kulthieth Formation) were sampled and fully characterized chemically and geologically. The coals have variable but high total organic carbon content technically classifying as coals and coaly shale, and highly varying PAH contents. Even for coals with high PAH content (approximately 4000 ppm total PAHs), a PAH-sensitive bacterial biosensor demonstrates nondetectable bioavailability as quantified, based on naphthalene as a test calibrant. These results are consistent with studies indicating that materials such as coals strongly diminish the bioavailability of hydrophobic organic compounds and support previous work suggesting that hydrocarbons associated with the regional background in northern Gulf of Alaska marine sediments are not appreciably bioavailable.

Seed biometric parameters in oil palm accessions from a Brazilian germplasm bank

The objective of this work was to evaluate the morphological diversity of oil palm seeds and to cluster the accessions according to their morphological characteristics. Forty-one accessions from the oil palm germplasm bank of Embrapa Amazônia Ocidental were evaluated - 18 of Elaeis oleifera and 23 of E. guineensis. The groups were formed based on morphological characteristics, by principal component analysis. In E. oleifera, four groups were formed, tied to their region of origin, but with significant morphological differences between accessions from the same population. For tenera-type E. guineensis seeds, three widely divergent groups were formed, especially as to external parameters, which differentiated them from the other ones. The parameter endocarp thickness stood out in intra- and inter-population differentiation. For dura-type E. guineensis, three groups were formed, with larger seeds and thicker endocarps, which differed from all the other ones. The variability observed for seed characteristics in the analyzed accessions allows the establishment of different groups, to define strategies for genetic improvement.