Interleaving Planning and Robot Execution for Asynchronous User Requests

ROGUE is an architecture built on a real robot which provides algorithms for the integration of high-level planning, low-level robotic execution, and learning. ROGUE addresses successfully several of the challenges of a dynamic office gopher environment. This article presents the techniques for the integration of planning and execution.ROGUE uses and extends a classical planning algorithm to create plans for multiple interacting goals introduced by asynchronous user requests. ROGUE translates the planner';s actions to robot execution actions and monitors real world execution. ROGUE is currently implemented using the PRODIGY4.0 planner and the Xavier robot. This article describes how plans are created for multiple asynchronous goals, and how task priority and compatibility information are used to achieve appropriate efficient execution. We describe how ROGUE communicates with the planner and the robot to interleave planning with execution so that the planner can replan for failed actions, identify the actual outcome of an action with multiple possible outcomes, and take opportunities from changes in the environment.ROGUE represents a successful integration of a classical artificial intelligence planner with a real mobile robot.     

Separation of non-sucrose compounds from sugar-beet syrup by ultrafiltration with ceramic membrane containing static mixer

The aim of this work was to investigate possible improvement of non-sucrose compounds separation from the syrup of raw brown sugar by application of a static mixer in an ultrafiltration process. The static mixer was expected to reduce the concentration polarization and fouling of the membrane. Non-affinated B sugar from the second stage of crystallization, diluted to 60°Bx dry matter, was used for preparing the solution subjected to the ultrafiltration. The cross-flow filtration, at a laboratory level, was carried out on the tubular ceramic membrane (Schumasiv Pall, USA), with a pore diameter of 5 nm. The separation was performed under various working conditions, with and without the presence of static mixer. The effect of turbulence promotion on filtration performances was investigated by using Kenics static mixer (FMX8124-AC, Omega). The process efficiency was quantified through the achieved values of the permeate flux, its colour and dry matter content, while the working factors were: fluid flow-rate, temperature, transmembrane pressure (TMP) and process duration. The positive effects of mixer application were proved.     

Evaluation of Adh1 alleles and transgenic soybean seeds using Scorpion PCR and HRM analysis

The identification of both approved and non-approved genetically modified organisms (GMOs) is an integral part of GMO biosafety legislation in many countries. One aspect that may affect PCR-based detection of a GMO lies within the analysis of its genetic stability, as sequence alterations or DNA instabilities may impede quantification by PCR. Genetic stability can be analyzed using various methods, yet many of these methods have distinct disadvantages, including low sensitivity. In this study, high resolution melting (HRM) analysis and real-time PCR with Scorpion primers were used as a method to analyze the 3′ end of RR soybeans (RR 40-3-2) in a large number of samples (n = 1,034). No evidence for the occurrence of mutation events was found, implying that the nucleotide sequence of this region is unlikely to be unstable and is well suited as a target for the quantification of RR soybeans. Additionally, and as a preparative work for an optimization of the method, a 174 bp region of the first intron of the Adh1 gene was analyzed in several varieties of maize with different GMO events using the same approach. The results show that 2 alleles are present. In further experiments, the different alleles were cloned into plasmids to generate homozygous plasmids from heterozygous templates in order to generate for a more precise analysis. The overall methodological aim of these studies was to compare HRM analysis with Scorpion primer PCR. Both methods were capable of differentiating between the 2 homozygous and heterozygous alleles. For a better discrimination, however, we conclude that it is most reliable to consider the results of both methods. This dual approach is assumed to be an effective tool as an accurate, high-throughput means of the screening of GMOs for potential genetic instabilities that may interfere with the detection and identification of specific GM events.     

Electrochemical method for the detection of lipase activity

A novel electrochemical technique for the general assay of lipase activity is described. The method utilizes a solid-supported lipase substrate, which is formed by dripping and drying a small amount of an ethanol solution of 9-(5'-ferrocenylpentanoyloxy)nonyl disulfide (FPONDS) onto gold modified by a hexanethiol self-assembled monolayer. The redox ferrocene group of FPONDS generates the electrochemical signal, the intensity of which is proportional to the number of FPONDS molecules at the interface. Electrochemical and surface-enhanced infrared absorption spectroscopic data, as well as control experiments with an engineered, deactivated mutant enzyme, demonstrate that the wild-type lipase from Thermomyces lanuginosus is capable of cleaving the ester bonds of FPONDS molecules via an enzymatic hydrolysis mechanism, which includes the adsorption of the lipase onto the substrate surface. The hydrolysis liberates the ferrocene groups from the interface triggering a decay of the electrochemical redox signal. The rate of the electrochemical signal decrease is proportional to the lipase activity/concentration. These data suggest a general method for the direct measure of enzymatic activity of lipases.     

Reduction of solid content in starch industry wastewater by microfiltration

Wastewater obtained in the production process of wheat starch and vital wheat gluten was treated by microfiltration through a ceramic tubular membrane with 200 nm pore sizes. The consumption of process water would thus be reduced, the starch would be recovered to a greater extent and the wastewater problem would consequently be solved. Reduction of the occurrence of polarization layer on the membrane and the constant wastewater permeate flux through the membrane was maintained by inserting of the mixer Kenics inside the membrane tube. The maximum value of the permeate flux (24 L m⁻² h⁻¹) without the use of a static mixer was achieved at 3 × 10⁻⁵ Pa and at a flow rate of 150 L/h, for wastewater samples initially allowed to settle for 4 h prior microfiltration. Under the very same conditions of the working parameters, the use of a static mixer enables a flux that is up to two or three times more intensive compared to the values obtained without using a mixer. Microfiltration reduces the wastewater dry matter from 11 000 to 4000 mg/L, lessens the chemical oxygen demand by 74%, i.e. from 21 000 to 5100 mgO₂/L and significantly decreases the values of the suspended matter, namely from 9000 to 300 mg/L.     

Chemical Approach to Biological Safety: Molecular‐Level Control of an Integrated Zinc Finger Nuclease

Application of artificial nucleases (ANs) in genome editing is still hindered by their cytotoxicity related to off‐target cleavages. This problem can be targeted by regulation of the nuclease domain. Here, we provide an experimental survey of computationally designed integrated zinc finger nucleases, constructed by linking the inactivated catalytic centre and the allosteric activator sequence of the colicin E7 nuclease domain to the two opposite termini of a zinc finger array. DNA specificity and metal binding were confirmed by electrophoretic mobility shift assays, synchrotron radiation circular dichroism spectroscopy, and nano‐electrospray ionisation mass spectrometry. In situ intramolecular activation of the nuclease domain was observed, resulting in specific cleavage of DNA with moderate activity. This study represents a new approach to AN design through integrated nucleases consisting of three (regulator, DNA‐binding, and nuclease) units, rather than simple chimera. The optimisation of such ANs could lead to safe gene editing enzymes.     

Sprouty3, but Not Sprouty1, Expression Is Beneficial for the Malignant Potential of Osteosarcoma Cells

Sprouty proteins are widely accepted modulators of receptor tyrosine kinase-associated pathways and fulfill diversified roles in cancerogenesis dependent on the originating cells. In this study we detected a high expression of Sprouty3 in osteosarcoma-derived cells and addressed the question of whether Sprouty3 and Sprouty1 influence the malignant phenotype of this bone tumor entity. By using adenoviruses, the Sprouty proteins were expressed in two different cell lines and their influence on cellular behavior was assessed. Growth curve analyses and Scratch assays revealed that Sprouty3 accelerates cell proliferation and migration. Additionally, more colonies were grown in Soft agar if the cells express Sprouty3. In parallel, Sprouty1 had no significant effect on the measured endpoints of the study in osteosarcoma-derived cells. The promotion of the tumorigenic capacities in the presence of Sprouty3 coincided with an increased activation of signaling as measured by evaluating the phosphorylation of extracellular signal-regulated kinases (ERKs). Ectopic expression of a mutated Sprouty3 protein, in which the tyrosine necessary for its activation was substituted, resulted in inhibited migration of the treated cells. Our findings identify Sprouty3 as a candidate for a tumor promoter in osteosarcoma.     

Size exclusion and reversed-phase high-performance liquid chromatography/UV for routine control of thermal processing of cows' and donkey milk major proteins

Cows' and donkey milks (raw and thermally processed) and respective whey were analysed for quantification of major proteins. Two different chromatographic approaches, size exclusion (SE-HPLC) and reversed-phase high performance liquid chromatography (RP-HPLC) both coupled to UV detection were used. Usefulness of these methods for routine control of the effect of thermal processing was evaluated. The external standard method was used to calibrate the SE-HPLC and RP-HPLC systems. Concerning quantification of β-lactoglobulin (β-lg), α-lactalbumin (α-la), lysozyme (lys), and total casein (cn), no significant differences between results obtained by SE-HPLC and by RP-HPLC (t-test, P>0·05) were observed for raw milks and whey. Heating of cows' milk promoted aggregation of denatured proteins as observed by SE-HPLC, whereas α-la and β-lg from donkey milk were stable to thermal processing at 100 °C (5 min). Lys was quantified in donkey raw milk and whey however, in thermally processed donkey milk lys was denatured and could not be quantified by HPLC.