NMR metabolomics of planktonic and biofilm modes of growth in Pseudomonas aeruginosa

Bacteria often reside in communities where the cells have secreted sticky, polymeric compounds that allow them to attach to surfaces. This sessile lifestyle, referred to as a biofilm, affords the cells within these communities a tolerance of antibiotics and antimicrobial treatments. Biofilms of the bacterium Pseudomonas aeruginosa have been implicated in cystic fibrosis and are capable of colonizing medical implant devices, such as heart valves and catheters, where treatment of the infection often requires the removal of the infected device. This mode of growth is in stark contrast to planktonic, free floating cells, which are more easily eradicated with antibiotics. The mechanisms contributing to a biofilm's tenacity and a planktonic cell's susceptibility are just beginning to be explored. In this study, we have used a metabolomic approach employing nuclear magnetic resonance (NMR) techniques to study the metabolic distinctions between these two modes of growth in P. aeruginosa. One-dimensional 1H NMR spectra of fresh growth medium were compared with spent medium supernatants from batch and chemostat planktonic and biofilms generated in continual flow system culture. In addition, 1H high-resolution magic angle spinning NMR techniques were employed to collect 1H NMR spectra of the corresponding cells. Principal component analysis and spectral comparisons revealed that the overall metabolism of planktonic and biofilm modes of growth appeared similar for the spent media, while the planktonic and biofilm cells displayed marked differences. To determine the robustness of this technique, we prepared cell samples under slightly different preparation methods. Both techniques showed similar results. These feasibility studies show that there exist chemical differences between planktonic and biofilm cells; however, in order to identify these metabolomic differences, more extensive studies would have to be performed, including 1H-1H total correlated spectroscopy.     

Continuous flow immobilized enzyme reactor-tandem mass spectrometry for screening of AChE inhibitors in complex mixtures

A method is described for identifying bioactive compounds in complex mixtures based on the use of capillary-scale monolithic enzyme-reactor columns for rapid screening of enzyme activity. A two-channel nanoLC system was used to continuously infuse substrate coupled with automated injections of substrate/small molecule mixtures, optionally containing the chromogenic Ellman reagent, through sol-gel derived acetylcholinesterase (AChE) doped monolithic columns. This is the first report of AChE encapsulated in monolithic silica for use as an immobilized enzyme reactor (IMER), and the first use of such IMERs for mixture screening. AChE IMER columns were optimized to allow rapid functional screening of compound mixtures based on changes in the product absorbance or the ratio of mass spectrometric peaks for product and substrate ions in the eluent. The assay had robust performance and produced a Z' factor of 0.77 in the presence of 2% (v/v) DMSO. A series of 52 mixtures consisting of 1040 compounds from the Canadian Compound Collection of bioactives was screened and two known inhibitors, physostigmine and 9-aminoacridine, were identified from active mixtures by manual deconvolution. The activity of the compounds was confirmed using the enzyme reactor format, which allowed determination of both IC(50) and K(I) values. Screening results were found to correlate well with a recently published fluorescence-based microarray screening assay for AChE inhibitors.     

The implications of household greywater treatment and reuse for municipal wastewater flows and micropollutant loads

An increasing worldwide interest in water recycling technologies such as greywater treatment and reuse suggests that additional research to elucidate the fate of xenobiotics during such practices would be beneficial. In this paper, scenario analyses supported by empirical data are used for highlighting the potential fate of a selection of xenobiotic micropollutants in decentralised greywater treatment systems, and for investigation of the possible implications of greywater recycling for the wider urban water cycle. Potential potable water savings of up to 43% are predicted for greywater recycling based on Danish water use statistics and priority substance monitoring at a greywater treatment plant in Denmark. Adsorption represents an important mechanism for the removal of cadmium, nickel, lead and nonylphenol from influent greywater and therefore the disposal route adopted for the generated sludge can exert a major impact on the overall efficiency and environmental sustainability of greywater treatment.     

Improved method for the determination of wax esters in vegetable oils

In this work, a modified International Olive Council (IOC) method for wax determination involving a double‐adsorbent layer of silica gel and silver nitrate‐impregnated silica gel is presented (SN method). Column chromatography by the SN method did not show retention of wax esters standards with an even number of carbon atoms (C34–C44), observing recovery percentages higher than 90% even for unsaturated wax esters. All wax fractions were lower by the SN method than by the IOC method, resulting in a percentage decrease in the total wax content (olive oils: 20–50%, crude sunflower oil: 38%, crude soybean oil: 58% and crude grape seed oil: 13%). Olive oils analysed by the SN method showed increases of up to 27% in C40 relative percentage with respect to the IOC method. Additionally, decreases were observed by the SN method in the relative percentages for odd‐carbon atom waxes for the seed oils in comparison to the IOC method (crude sunflower oil: 27%, crude soybean oil: 28% and crude grape seed oil: 13%). The main advantages of the proposed modification consist in its easy implementation and a better determination of wax esters (C34–C60) by controlling their complete recovery and removing interfering substances. The method is suitable for quality control and for authentication of olive oil and seed oils as well as in processing monitoring. Practical applications: The proposed method is useful in the quality, authentication and processing control of fruit and seed oils. Moreover, it can be an important tool for vegetable oil industries to control the efficiency of the wax separation process to prevent turbidity in the refined oil.     

Imbalance between phytoplankton production and bacterial carbon demand in relation to mucilage formation in the Northern Adriatic Sea

Spatial and temporal changes in phytoplankton production and bacterial C demand were investigated at four stations in the Northern Adriatic Sea over 3 years. The effect of the Po River plume was observed at the western stations; in particular, the northernmost one (B06) showed the highest values of primary production, both as hourly peaks (up to 14 mg C m(-3) h(-1)) and daily water column integrated values (up to 740 mg C m(-2) day(-1)), the southern station (C04) was only sporadically influenced and did not differ significantly from the easternmost ones (C12 and B13), where the lowest TPP values were recorded (around 1 mg C m(-3) h(-1)). In this study the first in situ data are reported on short-term phytoplankton C extra cellular release in the Northern Adriatic Sea. At every station a considerable percentage of primary production (PER>20% as an average, with peaks of up to 70%) was released as dissolved organic carbon. In particular, an association of fairly high PER (>10%) and specific production (Pb>10 mg C mg chl(-1) h(-1)) was observed from spring to summer, when the mucilage phenomenon usually starts. This result might suggest the presence of an uncoupling between photosynthesis and growth, probably related with nutrient availability, which would be responsible for a high production of extra cellular organic carbon. Phytoplankton primary production and bacterial carbon production were closely related and bacterial C production accounted, on average, for a higher percentage of primary production than the values typically reported in the literature on aquatic environments. The flow of organic matter from phytoplankton to bacteria seems to satisfy the bacterial carbon demand during most of the spring and summer, at least in the upper water layers. However, during the summer, there is evidence that BCD sometimes exceeds the amount of C produced by phytoplankton. Neither phytoplankton nor bacterial production showed significant differences over the relevant years, and their absolute values did not change when comparing periods with or without mucilage. However, there were indications of an uncoupling between phytoplankton photosynthesis and growth and of a shift from an autotrophic to a heterotrophic metabolism, especially during the spring and summer period when mucilage might occur.     

Analogical reasoning in a capuchin monkey (Cebus apella).

Previous evidence has suggested that analogical reasoning (recognizing similarities among object relations when the objects themselves are dissimilar) is limited to humans and apes. This study investigated whether capuchin monkeys (Cebus apella) can use analogical reasoning to solve a 3-dimensional search task. The task involved hiding a food item under 1 of 2 or 3 plastic cups of different sizes and then allowing subjects to search for food hidden under the cup of analogous size in their own set of cups. Four monkeys were exposed to a series of relational matching tasks. If subjects reached criterion on these tasks, they were exposed to relational transfer tasks involving novel stimuli. Three of the monkeys failed to reach criterion on the basic relational matching tasks and therefore were not tested further. One monkey, however, revealed above-chance performance on a series of transfer tasks with 3 novel stimuli. This evidence suggests that contrary to previous arguments, a member of a New World monkey species can solve an analogical problem. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Macroindentation of polymers

Macroindentation tests were performed by means of a flat cylindrical indenter on high‐density polyethylene (HDPE) and polyamide 66 (PA66). Commercial sheets were acquired for both materials, and specimens were extracted for differential scanning calorimetry (DSC), dynamic mechanical analysis (DMA), and thermal recovery tests. In this way, the fabrication effect was correctly identified in terms of degradation, orientation, and mechanical performance. Specimens were also prepared for tensile tests, which were performed for comparison with indentation experiments. Indentation tests were carried out on the external sheet surfaces at different indenter diameters (1 and 2 mm) and test rates (from 0.1 to 20 mm/min). A very high correlation was found between the tensile test and indentation results at the different test rates. In particular, the indentation stress at fixed depth was a linear function of the tensile yield stress, depending only on the indenter diameter and not on the material type. Moreover, a threshold value for the thickness for a correct comparison among samples having different dimensions is discussed. POLYM. ENG. SCI., 2008. © 2008 Society of Plastics Engineers     

Concentration of soy protein isolate affects starch-based confections' texture, sensory, and storage properties

The effects of increasing soy protein isolate concentration on the physico-chemical properties of starch-based grape confectionery gels were investigated using thermal, textural, and sensory analyses. Soy protein isolate decreased hardness, cohesiveness, and gumminess, demonstrating potential as a texture modifier. Increasing soy protein concentration progressively decreased the elastic properties of the starch network demonstrated by a lower G'-G" crossover frequency. High levels of soy protein also created a more homogeneous water population, one which was lost at lower temperatures compared to standard confections. An improvement in taste and texture acceptability of the confections upon addition of soy protein isolate was found by sensory analysis. Finally, physico-chemical properties were compared up to 20 d of storage at ambient temperature. Soy-containing confections demonstrated lower gumminess and cohesiveness during the entire storage period but a significant increase in hardness after 20 d of storage. PRACTICAL APPLICATION: Soy protein isolate imparted soft texture to starch-based confections on a concentration-dependent basis. Increasing soy protein isolate concentration decreased elastic properties of the starch network in starch confections and significantly improved flavor and texture. Gumminess and cohesiveness of soy protein isolate confection was lower during 20 d of storage.     

Detection of heavy metal ions in drinking water using a high-resolution differential surface plasmon resonance sensor

We have built a high-resolution differential surface plasmon resonance (SPR) sensor for heavy metal ion detection. The sensor surface is divided into a reference and sensing areas, and the difference in the SPR angles from the two areas is detected with a quadrant cell photodetector as a differential signal. In the presence of metal ions, the differential signal changes due to specific binding of the metal ions onto the sensing area coated with properly selected peptides, which provides an accurate real-time measurement and quantification of the metal ions. Selective detection of Cu2+ and Ni2+ in the ppt-ppb range was achieved by coating the sensing surface with peptides NH2-Gly-Gly-His-COOH and NH2-(His)6-COOH. Cu2+ in drinking water was tested using this sensor.