Plasmid DNA partitioning and separation using poly(ethylene glycol)/poly(acrylate)/salt aqueous two-phase systems

Phase diagrams of poly(ethylene glycol)/polyacrylate/Na2SO4 systems have been investigated with respect to polymer size and pH. Plasmid DNA from Escherichia coil can depending on pH and polymer molecular weight be directed to a poly(ethylene glycol) or to a polyacrylate-rich phase in an aqueous two-phase system formed by these polymers. Bovine serum albumin (BSA) and E. coil homogenate proteins can be directed opposite to the plasmid partitioning in these systems. Two bioseparation processes have been developed where in the final step the pDNA is partitioned to a salt-rich phase giving a total process yield of 60-70%. In one of them the pDNA is partitioned between the polyacrylate and PEG-phases in order to remove proteins. In a more simplified process the plasmid is partitioned to a PEG-phase and back-extracted into a Na2SO4-rich phase. The novel polyacrylate/PEG system allows a strong change of the partitioning between the phases with relatively small changes in composition or pH. (C) 2012 Elsevier B.V. All rights reserved.

Social facilitation and food partitioning in the queenless ant Dinoponera quadriceps (Hymenoptera: Formicidae)

Social facilitation occurs when an animal is more likely to behave in a certain way in response to other animals engaged in the same behaviour. For example, an individual returning to the nest with food stimulates other ants to leave and to forage. In the present study we demonstrate the existence of new facets in the colony organization of Dinoponera quadriceps: a positive feedback between the incoming food and the activation of new foragers, and the occurrence of incipient task partitioning during the food sharing. Lower-ranked workers located inside the nest process protein resources and higher-ranked workers handle smaller pieces and distribute them to the larvae. In conclusion, D. quadriceps has a decentralized pattern of task allocation with a double regulatory mechanism, which can be considered a sophisticated aspect of division of labour in ponerine ants.

Partitioning and extraction of collagenase from Penicillium aurantiogriseum in poly(ethylene glycol)/phosphate aqueous two-phase system

Purification of collagenase produced by Penicillium aurantiogriseum URM4622 was carried using a PEG/phosphate aqueous two-phase system (ATPS). A 2(3)-full experimental design was used to investigate the influence of PEG molar mass, PEG concentration and phosphate concentration on the selected responses, namely partition coefficient, activity yield and purification factor. The ATPS was composed of PEG (molar mass of 550, 1500 and 4000 g/mol) at concentrations of 15.0, 17.5 and 20.0% (w/w) and phosphate at concentrations of 12.5, 15.0 and 17.5% (w/w). The best results of one-step extraction of collagenase from the fermentation broth (partition coefficient of 1.01, activity yield of 242% and purification factor of 23.5) were obtained at pH 6.0 using 20.0% (w/w) PEG 550 and 17.5% (w/w) phosphate. The results of this preliminary study demonstrate that the selected ATPS is satisfactorily selective for the extraction of such a collagenase. (C) 2012 Elsevier B.V. All rights reserved.

Stand-level patterns of carbon fluxes and partitioning in a Eucalyptus grandis plantation across a gradient of productivity, in Sao Paulo State, Brazil

Wood production represents a large but variable fraction of gross primary production (GPP) in highly productive Eucalyptus plantations. Assessing patterns of carbon (C) partitioning (C flux as a fraction of GPP) between above- and belowground components is essential to understand mechanisms driving the C budget of these plantations. Better knowledge of fluxes and partitioning to woody and non-woody tissues in response to site characteristics and resource availability could provide opportunities to increase forest productivity. Our study aimed at investigating how C allocation varied within one apparently homogeneous 90 ha stand of Eucalyptus grandis (W. Hill ex Maiden) in Southeastern Brazil. We assessed annual above-ground net primary production (ANPP: stem, leaf, and branch production) and total belowground C flux (TBCF: the sum of root production and respiration and mycorrhizal production and respiration), GPP (computed as the sum of ANPP, TBCF and estimated aboveground respiration) on 12 plots representing the gradient of productivity found within the stand. The spatial heterogeneity of topography and associated soil attributes across the stand likely explained this fertility gradient. Component fluxes of GPP and C partitioning were found to vary among plots. Stem NPP ranged from 554 g C m(-2) year(-1) on the plot with lowest GPP to 923 g C m(-2) year(-1) on the plot with highest GPP. Total belowground carbon flux ranged from 497 to 1235 g C m(-2) year(-1) and showed no relationship with ANPP or GPP. Carbon partitioning to stem NPP increased from 0.19 to 0.23, showing a positive trend of increase with GPP (R-2 = 0.29, P = 0.07). Variations in stem wood production across the gradient of productivity observed at our experimental site were a result of the variability in C partitioning to different forest system components.

Brazilian sardine (Sardinella brasiliensis Steindachner, 1879) spawning and nursery habitats: spatial-scale partitioning and multiscale relationships with thermohaline descriptors

We provide a detailed account of the spatial structure of the Brazilian sardine (Sardinella brasiliensis) spawning and nursery habitats, using ichthyoplankton data from nine surveys (1976-1993) covering the Southeastern Brazilian Bight (SBB). The spatial variability of sardine eggs and larvae was partitioned into predefined spatial-scale classes (broad scale, 200-500 km; medium scale, 50-100 km; and local scale, <50 km). The relationship between density distributions at both developmental stages and environmental descriptors (temperature and salinity) was also explored within these spatial scales. Spatial distributions of sardine eggs were mostly structured on medium and local scales, while larvae were characterized by broad-and medium-scale distributions. Broad-and medium-scale surface temperatures were positively correlated with sardine densities, for both developmental stages. Correlations with salinity were predominantly negative and concentrated on a medium scale. Broad-scale structuring might be explained by mesoscale processes, such as pulsing upwelling events and Brazil Current meandering at the northern portion of the SBB, while medium-scale relationships may be associated with local estuarine outflows. The results indicate that processes favouring vertical stability might regulate the spatial extensions of suitable spawning and nursery habitats for the Brazilian sardine.

Is shell partitioning between the hermit crabs Pagurus brevidactylus and Pagurus criniticornis explained by interference and/or exploitation competition?

The effect of crab behaviour on shell-use dynamics was analysed, comparing both interference and exploitation behaviours between the hermit crabs Pagurus criniticornis and Pagurus brevidactylus. Although these species exhibited microhabitat separation, with P. criniticornis dominating (100%) in sandy substrates and P. brevidactylus (80%) on rocky shores, they overlapped in the rocky shore/sand interface (P. criniticornis, 53%; P. brevidactylus, 43%). Pagurus criniticornis occupied shells of Cerithium atratum in higher frequencies (84%) than P. brevidactylus (37%), which was hypothesized to be a consequence of competitive interactions combined with their ability to acquire and/or retain this resource. The species P. criniticornis was attracted in larger numbers to simulated gastropod predation events than was P. brevidactylus, which, on the few occasions that it moved before P. criniticornis, tended to be attracted more rapidly. Interspecific shell exchanges between these species were few, suggesting the absence of dominance relationships. The shell-use pattern in this species pair is thus defined by exploitation competition, which is presumed to be intensified in areas of microsympatry. These results differ from other studies, which found that interference competition through interspecific exchanges shapes shell use, indicating that shell partitioning in hermit crabs is dependent on the behaviour of the species involved in the contests.

Soil carbon balance in a tropical grassland: Estimation of soil respiration and its partitioning using a semi-empirical model

In savannah and tropical grasslands, which account for 60% of grasslands worldwide, a large share of ecosystem carbon is located below ground due to high root:shoot ratios. Temporal variations in soil CO2 efflux (R-S) were investigated in a grassland of coastal Congo over two years. The objectives were (1) to identify the main factors controlling seasonal variations in R-S and (2) to develop a semi-empirical model describing R-S and including a heterotrophic component (R-H) and an autotrophic component (R-A). Plant above-ground activity was found to exert strong control over soil respiration since 71% of seasonal R-S variability was explained by the quantity of photosynthetically active radiation absorbed (APAR) by the grass canopy. We tested an additive model including a parameter enabling R-S partitioning into R-A and R-H. Assumptions underlying this model were that R-A mainly depended on the amount of photosynthates allocated below ground and that microbial and root activity was mostly controlled by soil temperature and soil moisture. The model provided a reasonably good prediction of seasonal variations in R-S (R-2 = 0.85) which varied between 5.4 mu mol m(-2) s(-1) in the wet season and 0.9 mu mol m(-2) s(-1) at the end of the dry season. The model was subsequently used to obtain annual estimates of R-S, R-A and R-H. In accordance with results reported for other tropical grasslands, we estimated that R-H accounted for 44% of R-S, which represented a flux similar to the amount of carbon brought annually to the soil from below-ground litter production. Overall, this study opens up prospects for simulating the carbon budget of tropical grasslands on a large scale using remotely sensed data. (C) 2012 Elsevier B.V. All rights reserved.

Performance evaluation of asphalt pavements using the Incremental-Recursive procedure of the Mechanistic-Empirical design software CalME

Il concetto di “sostenibilità” si riferisce allo sviluppo dei sistemi umani attraverso il più piccolo impatto possibile sul sistema ambientale. Le opere che si inseriscono bene nel contesto ambientale circostante e le pratiche che rispettano le risorse in maniera tale da permettere una crescita e uno sviluppo a lungo termine senza impattare sull’ambiente sono indispensabili in una società moderna. I progressi passati, presenti e futuri che hanno reso i conglomerati bituminosi materiali sostenibili dal punto di vista ambientale sono particolarmente importanti data la grande quantità di conglomerato usato annualmente in Europa e negli Stati Uniti. I produttori di bitume e di conglomerato bituminoso stanno sviluppando tecniche innovative per ridurre l’impatto ambientale senza compromettere le prestazioni meccaniche finali. Un conglomerato bituminoso ad “alta lavorabilità” (WMA), pur sviluppando le stesse caratteristiche meccaniche, richiede un temperatura di produzione minore rispetto a quella di un tradizionale conglomerato bituminoso a caldo (HMA). L’abbassamento della temperature di produzione riduce le emissioni nocive. Questo migliora le condizioni dei lavoratori ed è orientato verso uno sviluppo sostenibile. L’obbiettivo principale di questa tesi di laurea è quello di dimostrare il duplice valore sia dal punto di vista dell’eco-compatibilità sia dal punto di vista meccanico di questi conglomerati bituminosi ad “alta lavorabilità”. In particolare in questa tesi di laurea è stato studiato uno SMA ad “alta lavorabilità” (PGGWMA). L’uso di materiali a basso impatto ambientale è la prima fase verso un progetto ecocompatibile ma non può che essere il punto di partenza. L’approccio ecocompatibile deve essere esteso anche ai metodi di progetto e alla caratterizzazione di laboratorio dei materiali perché solo in questo modo è possibile ricavare le massime potenzialità dai materiali usati. Un’appropriata caratterizzazione del conglomerato bituminoso è fondamentale e necessaria per una realistica previsione delle performance di una pavimentazione stradale. La caratterizzazione volumetrica (Mix Design) e meccanica (Deformazioni Permanenti e Comportamento a fatica) di un conglomerato bituminoso è una fase importante. Inoltre, al fine di utilizzare correttamente i materiali, un metodo di progetto avanzato ed efficiente, come quello rappresentato da un approccio Empirico-Meccanicistico (ME), deve essere utilizzato. Una procedura di progetto Empirico-Meccanicistica consiste di un modello strutturale capace di prevedere gli stati di tensione e deformazione all’interno della pavimentazione sotto l’azione del traffico e in funzione delle condizioni atmosferiche e di modelli empirici, calibrati sul comportamento dei materiali, che collegano la risposta strutturale alle performance della pavimentazione. Nel 1996 in California, per poter effettivamente sfruttare i benefici dei continui progressi nel campo delle pavimentazioni stradali, fu iniziato un estensivo progetto di ricerca mirato allo sviluppo dei metodi di progetto Empirico - Meccanicistici per le pavimentazioni stradali. Il risultato finale fu la prima versione del software CalME che fornisce all’utente tre approcci diversi di l’analisi e progetto: un approccio Empirico, uno Empirico - Meccanicistico classico e un approccio Empirico - Meccanicistico Incrementale - Ricorsivo. Questo tesi di laurea si concentra sulla procedura Incrementale - Ricorsiva del software CalME, basata su modelli di danno per quanto riguarda la fatica e l’accumulo di deformazioni di taglio dai quali dipendono rispettivamente la fessurazione superficiale e le deformazioni permanenti nella pavimentazione. Tale procedura funziona per incrementi temporali successivi e, usando i risultati di ogni incremento temporale, ricorsivamente, come input dell’incremento temporale successivo, prevede le condizioni di una pavimentazione stradale per quanto riguarda il modulo complesso dei diversi strati, le fessurazioni superficiali dovute alla fatica, le deformazioni permanenti e la rugosità superficiale. Al fine di verificare le propreità meccaniche del PGGWMA e le reciproche relazioni in termini di danno a fatica e deformazioni permanenti tra strato superficiale e struttura della pavimentazione per fissate condizioni ambientali e di traffico, è stata usata la procedura Incrementale – Ricorsiva del software CalME. Il conglomerato bituminoso studiato (PGGWMA) è stato usato in una pavimentazione stradale come strato superficiale di 60 mm di spessore. Le performance della pavimentazione sono state confrontate a quelle della stessa pavimentazione in cui altri tipi di conglomerato bituminoso sono stati usati come strato superficiale. I tre tipi di conglomerato bituminoso usati come termini di paragone sono stati: un conglomerato bituminoso ad “alta lavorabilità” con granulometria “chiusa” non modificato (DGWMA), un conglomerato bituminoso modificato con polverino di gomma con granulometria “aperta” (GGRAC) e un conglomerato bituminoso non modificato con granulometria “chiusa” (DGAC). Nel Capitolo I è stato introdotto il problema del progetto ecocompatibile delle pavimentazioni stradali. I materiali a basso impatto ambientale come i conglomerati bituminosi ad “alta lavorabilità” e i conglomerati bituminosi modificati con polverino di gomma sono stati descritti in dettaglio. Inoltre è stata discussa l’importanza della caratterizzazione di laboratorio dei materiali e il valore di un metodo razionale di progetto delle pavimentazioni stradali. Nel Capitolo II sono stati descritti i diversi approcci progettuali utilizzabili con il CalME e in particolare è stata spiegata la procedura Incrementale – Ricorsiva. Nel Capitolo III sono state studiate le proprietà volumetriche e meccaniche del PGGWMA. Test di Fatica e di Deformazioni Permanenti, eseguiti rispettivamente con la macchina a fatica per flessione su quattro punti e il Simple Shear Test device (macchina di taglio semplice), sono stati effettuati su provini di conglomerato bituminoso e i risultati dei test sono stati riassunti. Attraverso questi dati di laboratorio, i parametri dei modelli della Master Curve, del danno a fatica e dell’accumulo di deformazioni di taglio usati nella procedura Incrementale – Ricorsiva del CalME sono stati valutati. Infine, nel Capitolo IV, sono stati presentati i risultati delle simulazioni di pavimentazioni stradali con diversi strati superficiali. Per ogni pavimentazione sono stati analizzati la fessurazione superficiale complessiva, le deformazioni permanenti complessive, il danno a fatica e la profondità delle deformazioni in ognuno degli stati legati.

Geological and hydrogeological features affecting migration, multi-phase partitioning and degradation of chlorinated hydrocarbons through unconsolidated porous media.

Chlorinated solvents are the most ubiquitous organic contaminants found in groundwater since the last five decades. They generally reach groundwater as Dense Non-Aqueous Phase Liquid (DNAPL). This phase can migrate through aquifers, and also through aquitards, in ways that aqueous contaminants cannot. The complex phase partitioning to which chlorinated solvent DNAPLs can undergo (i.e. to the dissolved, vapor or sorbed phase), as well as their transformations (e.g. degradation), depend on the physico-chemical properties of the contaminants themselves and on features of the hydrogeological system. The main goal of the thesis is to provide new knowledge for the future investigations of sites contaminated by DNAPLs in alluvial settings, proposing innovative investigative approaches and emphasizing some of the key issues and main criticalities of this kind of contaminants in such a setting. To achieve this goal, the hydrogeologic setting below the city of Ferrara (Po plain, northern Italy), which is affected by scattered contamination by chlorinated solvents, has been investigated at different scales (regional and site specific), both from an intrinsic (i.e. groundwater flow systems) and specific (i.e. chlorinated solvent DNAPL behavior) point of view. Detailed investigations were carried out in particular in one selected test-site, known as “Caretti site”, where high-resolution vertical profiling of different kind of data were collected by means of multilevel monitoring systems and other innovative sampling and analytical techniques. This allowed to achieve a deep geological and hydrogeological knowledge of the system and to reconstruct in detail the architecture of contaminants in relationship to the features of the hosting porous medium. The results achieved in this thesis are useful not only at local scale, e.g. employable to interpret the origin of contamination in other sites of the Ferrara area, but also at global scale, in order to address future remediation and protection actions of similar hydrogeologic settings.

Quadratically Hyponormal Weighted Shifts with Recursive Tail

We characterize positive quadratic hyponormality of the weighted shift W-alpha(x) associated to the weight sequence alpha(x) : 1, 1, root x, (root u, root v, root w)(Lambda) with Stampfli recursive tail, and produce an interval in x with non-empty interior in the positive real line for quadratic hyponormality but not positive quadratic hyponormality for such a shift. (C) 2013 Elsevier Inc. All rights reserved.