Kinetics and performance of a co-immobilised system of amyloglucosidase and Zymomonas mobilis.

High operational stability and productivity of co-immobilised systems are important aspects for their successful application in industrial processes. A dynamic model is required to describe artificially co-immobilised systems because the time needed to reach steady state normally exceeds the operational life span of these systems. Time dependent intraparticle concentration profiles and macroscopic conversion were modelled to study the operational stability and productivity of these systems theoretically. The model was used to describe experimental results of ethanol production from maltose by a co-immobilised system of amyloglucosidase and Zymomonas mobilis. Furthermore, the influence of the immobilisation procedure with glutaraldehyde and polyethyleneimine could also be studied with and incorporated in the model. From the model it could be derived that co-immobilised systems performing a consecutive reaction evolve towards a steady state, characterised by a constant concentration of the intermediate in the particle if product inhibition is neglected. Such a situation develops independently of the biomass concentration and the radial position, and has important consequences for co-immobilised systems. When the concentration of the intermediate in the bulk liquid is lower than this constant value in the biocatalyst particle, two regions may be distinguished in the particle: an inactive peripheral region without biomass and an active core with a biomass concentration depending on the substrate and immobilised enzyme concentration. Unlike immobilised single cell systems, it is possible to obtain a real steady state and therefore a stable situation for co-immobilised systems. However, a high operational life time could only be achieved at the expense of the productivity of the biocatalyst particle. A stability criterion is derived which agrees very well with the simulation results.     

Origin of the genetic code: first aminoacyl-tRNA synthetases could replace isofunctional ribozymes when only the second base of codons was established

Analysis of the updated compilation of more than 8,000 tRNA gene sequences confirmed our previously reported finding that in pairs of consensus tRNAs with complementary anticodons, their second bases in the acceptor stems are also complementary. This dual complementarity points to the following: (1) the operational code embodied in the acceptor stem, and the classic genetic code embodied in the anticodon could have had the same common ancestor; (2) new tRNAs most likely entered primitive translation in pairs with complementary anticodons; and (3) this process of code expansion was directed by the primordial double-strand coding. However, we did not find the dual complementarity when testing all tRNA pairs in which anticodons were complementary only at the central position, but not complementary at least at one of the flanking two positions. This observation, together with certain additional evidence, suggests that both codes were still being shaped (with only the second base established at the time) when the first protein aminoacyl-tRNA synthetases could have already started replacing their ribozymic precursors.     

Studies on the Natural Flavoux Components of Psidium gunajava L. Fruit

In the study Amberlite XAD-4 resin was employed as an adsorbent, which adsorbed the natural flavour sent forth from Psidium guajava L. fruit. The eluant of the adsorbate was concentrated by using a K-D concentrator. The concentrate was analyzed by dual capillary gas chromatography-mass spectrometry and temperature programed retention index on dual columns. A total of 25 substances were identified, which amounted to 98% of the total area of .the peaks. Four acetals were reported for the first time as guava components, i.e., l,l-dietho- xymethane, l,l-diethoxyethane, 1,1-diethoxyhexane and acetaldehyde ethyl cis-3-hexenyl acetal. Evaluation at an odour port during GC showed that the sharp characteristic aroma sent forth from guava fruitis attributed to l,l-diethoxymethane and l,l-diethoxyethane principally. It was discussed that the method of combining the dual column GC-MS with the dual column retention index could increase the reliability of the identification results.     

Continuous production of maltotetraose using a dual immobilized enzyme system of maltotetraose-forming amylase and pullulanase

In order to produce a product with a high content of maltotetraose, dual-enzyme systems composed of immobilized maltotetraose-forming amylase (G(4)-forming amylase) and pullulanase were studied. The thermostability of individually immobilized enzymes was examined in continuous operation; studies revealed that the enzyme immobilized on "Chitopearl" was much more stable than that immobilized on Diaion HP-50. The effects of operating conditions on the stability of G(4) forming amylase immobilized on "Chitopearl" were examined to confirm that the apparent half-life data could be arranged using the immobilized enzyme stability factor, f(s). As for the dual immobilized enzyme system, six methods of usage were considered, with five yielding a 7-10% (w/w) higher content of maltotetraose product than the single-enzyme system. The effects of operating conditions on the maltotetraose production reaction were examined to confirm that the maltotetraose content of the products could be analyzed using the specific space velocity,SSV. In dual immobilized enzyme systems, pullulanase immobilized on the same carrier as the G(4)-forming amylase was found to be more stable than pullulanase immobilized on separate carriers. The effectiveness of using immobilized pullulanase along with the G(4)-forming amylase was confirmed from constant-conversion operations in which the maltotetraose content in the product was kept at 50% (w/w) in laboratory-scale experimentation.     

The protective effect of breast feeding in relation to sudden infant death syndrome (SIDS): I. The effect of human milk and infant formula preparations on binding of toxigenic Staphylococcus aureus to epithelial cells

Epidemiological studies indicate that breast-fed infants are at a decreased risk of sudden infant death syndrome (SIDS) compared to formula-fed infants. Increasing evidence suggests that infectious agents might be involved in some of these deaths, in particular bacteria which colonise mucosal surfaces and produce superantigenic toxins. One species implicated in recent studies of SIDS infants is Staphylococcus aureus. We tested the hypothesis that in comparison to infant formula, human milk might be a better inhibitor of binding of S. aureus to epithelial cells. In this study, two protocols were used for the binding assays which were assessed by flow cytometry: the in vitro method in which bacteria were treated with milk or formula, washed and added to epithelial cells; and a method more closely reflecting the competitive interactions in vivo in which cells, bacteria, and milk or infant formula were added at the same time. With the in vivo method, breast milk caused enhancement of bacterial binding to cells whilst infant formula caused inhibition; however, for the in vitro method, both human milk and infant formula caused consistent enhancement of binding. Flow cytometry and light microscopy studies indicated that the enhancement was due to the formation of bacterial aggregates. Human milk and infant formula preparations were also compared for components (antibodies or oligosaccharides) that could inhibit binding of S. aureus using the in vitro method. Human milk contained both IgA and IgG. Neither human milk nor infant formula contained oligosaccharides reactive with the Ulex europaeus lectin but both contained components that bound monoclonal antibodies to Lewis(a) and Lewis(b) antigens which can act as receptors for S. aureus. With both methods, synthetic Lewis(a) and Lewis(b) inhibited S. aureus binding in a dose-dependent manner. With human milk, however, the only component which showed a significant correlation with inhibition of binding was the IgA specific for the staphylococcal surface component that binds Lewis(a). Both human milk and infant formula contain components which could potentially inhibit bacterial binding but only breast milk contains the IgA specific for the bacterial adhesin that binds Lewis(a). Studies using the in vivo method suggest that protection associated with breast feeding in relation to SIDS could be due mainly to the formation of bacterial aggregates. The studies have implications for further research into constituents of infant formula.     

An overview of energy efficiency techniques in cluster computing systems

Two major constraints demand more consideration for energy efficiency in cluster computing: (a) operational costs, and (b) system reliability. Increasing energy efficiency in cluster systems will reduce energy consumption, excess heat, lower operational costs, and improve system reliability. Based on the energy-power relationship, and the fact that energy consumption can be reduced with strategic power management, we focus in this survey on the characteristic of two main power management technologies: (a) static power management (SPM) systems that utilize low-power components to save the energy, and (b) dynamic power management (DPM) systems that utilize software and power-scalable components to optimize the energy consumption. We present the current state of the art in both of the SPM and DPM techniques, citing representative examples. The survey is concluded with a brief discussion and some assumptions about the possible future directions that could be explored to improve the energy efficiency in cluster computing.     

Systematic analysis of bicistronic reporter assay data

Bicistronic reporter assay systems have become a mainstay of molecular biology. While the assays themselves encompass a broad range of diverse and unrelated experimental protocols, the numerical data garnered from these experiments often have similar statistical properties. In general, a primary dataset measures the paired expression of two internally controlled reporter genes. The expression ratio of these two genes is then normalized to an external control reporter. The end result is a 'ratio of ratios' that is inherently sensitive to propagation of the error contributed by each of the respective numerical components. The statistical analysis of this data therefore requires careful handling in order to control for the propagation of error and its potentially misleading effects. A careful survey of the literature found no consistent method for the statistical analysis of data generated from these important and informative assay systems. In this report, we present a detailed statistical framework for the systematic analysis of data obtained from bicistronic reporter assay systems. Specifically, a dual luciferase reporter assay was employed to measure the efficiency of four programmed -1 frameshift signals. These frameshift signals originate from the L-A virus, the SARS-associated Coronavirus and computationally identified frameshift signals from two Saccharomyces cerevisiae genes. Furthermore, these statistical methods were applied to prove that the effects of anisomycin on programmed -1 frameshifting are statistically significant. A set of Microsoft Excel spreadsheets, which can be used as templates for data generated by dual reporter assay systems, and an online tutorial are available at our website (http://dinmanlab.umd.edu/statistics). These spreadsheets could be easily adapted to any bicistronic reporter assay system.     

Full-scale studies of factors related to coliform regrowth in drinking water.

An 18-month survey of 31 water systems in North America was conducted to determine the factors that contribute to the occurrence of coliform bacteria in drinking water. The survey included analysis of assimilable organic carbon (AOC), coliforms, disinfectant residuals, and operational parameters. Coliform bacteria were detected in 27.8% of the 2-week sampling periods and were associated with the following factors: filtration, temperature, disinfectant type and disinfectant level, AOC level, corrosion control, and operational characteristics. Four systems in the study that used unfiltered surface water accounted for 26.6% of the total number of bacterial samples collected but 64.3% (1,013 of 1,576) of the positive coliform samples. The occurrence of coliform bacteria was significantly higher when water temperatures were > 15 degrees C. For filtered systems that used free chlorine, 0.97% of 33,196 samples contained coliform bacteria, while 0.51% of 35,159 samples from chloraminated systems contained coliform bacteria. The average density of coliform bacteria was 35 times higher in free-chlorinated systems than in chloraminated water (0.60 CFU/100 ml for free-chlorinated water compared with 0.017 CFU/100 ml for chloraminated water). Systems that maintained dead-end free chlorine levels of < 0.2 mg/liter or monochloramine levels of < 0.5 mg/liter had substantially more coliform occurrences than systems that maintained higher disinfectant residuals. Free-chlorinated systems with AOC levels greater than 100 micrograms/liter had 82% more coliform-positive samples and 19 times higher coliform levels than free-chlorinated systems with average AOC levels less than 99 micrograms/liter. Systems that maintained a phosphate-based corrosion inhibitor and limited the amount of unlined cast iron pipe had fewer coliform bacteria. Several operational characteristics of the treatment process or the distribution system were also associated with increased rates of coliform occurrence. The study concludes that the occurrence of coliform bacteria within a distribution system is dependent upon a complex interaction of chemical, physical, operational, and engineering parameters. No one factor could account for all of the coliform occurrences, and one must consider all of the parameters described above in devising a solution to the regrowth problem.     

New structure in cell puncture activities by aphid stylets: a dual‐mode EPG study

Intracellular punctures by aphid stylets appear as potential drop (pd) waveforms in DC electrical penetration graph (EPG) recordings. We used a dual‐EPG device that recorded in one channel the ‘full EPG’ with R‐plus emf‐components (i.e., the usual DC EPG) and concurrently in a second channel the ‘R‐EPG’ with R‐components only. The circuit of the latter channel was an optimised amplitude modulation (AM) version derived from early (before 1990) AC systems. We also made some ‘emf‐EPG’ recordings using a separate high input resistance ‘emf‐amplifier’ sensitive to emf‐components only. The intracellular pd waveforms have previously been divided into three subphases, and we aimed to distinguish and separate these subphases more accurately by the dual‐EPG recordings than with the normal full EPG only. In this study, we temporarily distinguished five subphases (α–ε), but unequivocal distinction of only a few of these appeared possible, in spite of the information coming from the two signals. The lack of clearly separable features in R‐EPG signals often provided serious difficulties in pd recognition without the concurrent full EPG, but once located, only subphase II‐2 features were clear and supported the II‐2 data from the full EPG. Consequently, we could not distinguish subphases of complete pd waveforms better with additional R‐EPG information during cell punctures by Aphis gossypii Glover (Hemiptera: Aphididae). In Brevicoryne brassicae (L.) (Hemiptera: Aphididae), however, distinguishing II‐2 subphases in the full EPG was sometimes a problem. Our detailed dual‐EPG observations showed some waveform continuity from halfway into the II‐1 subphase (start of the newly recognised subphase β) until the end of the pd, with a strong but variable emf origin. This waveform tended to overrule other subphase waveforms in B. brassicae more than in A. gossypii and Myzus persicae (Sulzer) (Hemiptera: Aphididae). Subphase waveforms in full EPGs were especially difficult to recognise when pd periods had been interrupted in a virus inoculation experiment and additional R‐EPG information could then be useful. This inoculation experiment showed again that only the first subphase (II‐1) contributes to virus (Cucumber mosaic virus) inoculation by A. gossypii. In B. brassicae, the benefit of concurrent R‐EPG information in such virus experiments is presently under further investigation. Apart from this special application to virus experiments, we do not recommend the routine use of the dual‐EPG device. Furthermore, we do not advocate the distinction of more than the previously recognised three intracellular pd subphases as a feasible option in future studies. Analysis of EPGs with concurrent R‐EPGs requires substantially more analysis work without yielding consistently useful additional insights. This confirms earlier dual‐EPG results from thrips.     

Linkage between root systems of three pioneer plant species and soil nitrogen during early reclamation of a mine site in Lusatia,Germany

In 2005, a 7‐ha artificial watershed (Chicken Creek) was built on a post mined landscape in Lusatia, Germany from sandy substrates of Pleistocene origin, commonly used in reclamation. The watershed was developed to investigate the initial phase of soil and ecosystem development under natural conditions. At this early stage, mineral nitrogen in young sandy soils is primarily limited and nitrogen fixing legumes become key components of natural succession. Local abundant pioneering legumes Lotus corniculatus and Trifolium arvense and one pioneer grass species Calamagrostis epigeios were investigated 5 years after watershed construction. In this study, we investigated the influence of spatial root and nodule distribution of these species on soil nitrogen accumulation. Soil, including roots, was sampled from field monoliths covered with the aforementioned plant species. Root systems of both legumes were mainly restricted to the upper 20 cm of soil, whereas roots of C. epigeios also developed strongly at greater depths. A positive relationship was found, with higher plant densities associated with higher root densities which were associated with higher nodule densities for legumes and which were all associated with significantly higher soil nitrogen content relative to non‐vegetated areas. This research provides rare information on the role root systems of pioneer legumes play in soil nitrogen input in the early stage of soil and ecosystem development during revegetation by natural succession.