The evolution of a Pluto-like system during the migration of the ice giants

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Analysis of hourly global, direct and diffuse solar radiations attenuation as a function of optical air mass

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Reliability and differentiation capability of dynamic and static kinematic measurements of rearfoot eversion in patellofemoral pain

Excessive rearfoot eversion is thought to be a risk factor for patellofemoral pain development, due to the kinesiological relationship with ascendant adaptations. Individuals with patellofemoral pain are often diagnosed through static clinical tests, in scientific studies and clinical practice. However, the adaptations seem to appear in dynamic conditions. Performing static vs. dynamic evaluations of widely used measures would add to the knowledge in this area. Thus, the aim of this study was to determine the reliability and differentiation capability of three rearfoot eversion measures: rearfoot range of motion, static clinical test and static measurement using a three-dimensional system. A total of 29 individuals with patellofemoral pain and 25 control individuals (18-30 years) participated in this study. Each subject underwent three-dimensional motion analysis during stair climbing and static clinical tests. Intraclass correlation coefficient and standard error measurements were performed to verify the reliability of the variables and receiver operating characteristic curves to show the diagnostic accuracy of each variable. In addition, analyses of variance were performed to identify differences between groups. Rearfoot range of motion demonstrated higher diagnostic accuracy (an area under the curve score of 0.72) than static measures and was able to differentiate the groups. Only the static clinical test presented poor and moderate reliability. Other variables presented high to very high values. Rearfoot range of motion was the variable that presented the best results in terms of reliability and differentiation capability. Static variables do not seem to be related to patellofemoral pain and have low accuracy values.

Estudo fitoquímico dos extratos de folhas, galhos e cascas do caule de Calycophyllum spruceanum Benth para testes de potencial de cosmético funcional

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Compostos de paládio(II) contendo ligantes N,S-quelantes: síntese, caracterização e investigação da atividade biológica

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Impact of biannual rossby waves on the Indian Ocean Dipole

Topex/Poseidon sea surface height anomalies during 1993-2002 are decomposed using 2-D finite impulse response filters which showed biannual Rossby waves (BRWs) in the equatorial Indian Ocean (peak at 1.5 degrees S) and in the southern tropical Indian Ocean (peak at 10.5 degrees S) during Indian Ocean Dipole (IOD) years. Anomalous downwelling BRWs in the equatorial Indian Ocean triggered by the wind stress curl-induced Ekman pumping near the eastern boundary started propagating westward from the eastern boundary in July/August 1993 and 1996, i.e., more than one year prior to the formation of the IOD events of 1994 and 1997 respectively. These strong downwelling signals reach the western equatorial Indian Ocean during the peak dipole time.

Revealing archaeal diversity patterns and methane fluxes in Admiralty Bay, King George Island, and their association to Brazilian Antarctic Station activities

The study of Antarctic archaeal communities adds information on the biogeography of this group and helps understanding the dynamics of biogenic methane production in such extreme habitats. Molecular methods were combined to methane flux determinations in Martel Inlet, Admiralty Bay, to assess archaeal diversity, to obtain information about contribution of the area to atmospheric methane budget and to detect possible interferences of the Antarctic Brazilian Station Comandante Ferraz (EACF) wastewater discharge on local archaeal communities and methane emissions. Methane fluxes in Martel Inlet ranged from 3.2 to 117.9 mu mol CH(4) m(-2) d(-1), with an average of 51.3 +/- 8.5 mu mol CH(4) m(-2) d(-1) and a median of 57.6 mu mol CH(4) m(-2)d(-1). However, three negative fluxes averaging -11.3 mu mol CH(4) m(-2) d(-1) were detected in MacKellar Inlet, indicating that Admiralty Bay can be either a source or sink of atmospheric methane. Denaturing gradient gel electrophoresis (DGGE) showed that archaeal communities at EACF varied with depth and formed a group separated from the reference sites. Granulometric analysis indicated that differences observed may be mostly related to sediment type. However, an influence of wastewater input could not be discarded, since higher methane fluxes were found at CF site. suggesting stimulation of local methanogenesis. DGGE profile of the wastewater sample grouped separated from all other samples, suggesting that methanogenesis stimulation may be due to changes in environmental conditions rather than to the input of allochtonous species from the wastewater. 16S ribosomal DNA clone libraries analysis showed that all wastewater sequences were related to known methanogenic groups belonging to the hydrogenotrophic genera Methanobacterium and Methanobrevibacter and the aceticlastic genus Methanosaeta. EACF and Botany Point sediment clone libraries retrieved only groups of uncultivated Archaea, with predominance of Crenarchaeota representatives (MCG, MG1, MBG-B, MBG-C and MHVG groups). Euryarchaeota sequences found were mostly related to the LDS and RC-V groups, but MBG-D and DHVE-5 were also present. No representatives of cultivated methanogenic groups were found, but coverage estimates suggest that a higher number of clones would have to be analyzed in order to cover the greater archaeal diversity of Martel Inlet sediment. Nevertheless, the analysis of the libraries revealed groups not commonly found by other authors in Antarctic habitats and also indicated the presence of groups of uncultivated archaea previously associated to methane rich environments or to the methane cycle. (C) 2010 Elsevier Ltd. All rights reserved.

Methane airborne measurements and comparison to global models during BARCA

Tropical regions, especially the Amazon region, account for large emissions of methane (CH4). Here, we present CH4 observations from two airborne campaigns conducted within the BARCA (Balanco Atmosferico Regional de Carbono na Amazonia) project in the Amazon basin in November 2008 (end of the dry season) and May 2009 (end of the wet season). We performed continuous measurements of CH4 onboard an aircraft for the first time in the Amazon region, covering the whole Amazon basin with over 150 vertical profiles between altitudes of 500 m and 4000 m. The observations support the finding of previous ground-based, airborne, and satellite measurements that the Amazon basin is a large source of atmospheric CH4. Isotope analysis verified that the majority of emissions can be attributed to CH4 emissions from wetlands, while urban CH4 emissions could be also traced back to biogenic origin. A comparison of five TM5 based global CH4 inversions with the observations clearly indicates that the inversions using SCIAMACHY observations represent the BARCA observations best. The calculated CH4 flux estimate obtained from the mismatch between observations and TM5-modeled CH4 fields ranges from 36 to 43 mg m(-2) d(-1) for the Amazon lowland region.

Probing the cosmic distance-duality relation with the Sunyaev-Zel'dovich effect, X-ray observations and supernovae Ia

Context. The angular diameter distances toward galaxy clusters can be determined with measurements of Sunyaev-Zel'dovich effect and X-ray surface brightness combined with the validity of the distance-duality relation, D-L(z)(1 + z)(2)/D-A(z) = 1, where D-L(z) and D-A(z) are, respectively, the luminosity and angular diameter distances. This combination enables us to probe galaxy cluster physics or even to test the validity of the distance-duality relation itself. Aims. We explore these possibilities based on two different, but complementary approaches. Firstly, in order to constrain the possible galaxy cluster morphologies, the validity of the distance-duality relation (DD relation) is assumed in the Lambda CDM framework (WMAP7). Secondly, by adopting a cosmological-model-independent test, we directly confront the angular diameters from galaxy clusters with two supernovae Ia (SNe Ia) subsamples (carefully chosen to coincide with the cluster positions). The influence of the different SNe Ia light-curve fitters in the previous analysis are also discussed. Methods. We assumed that eta is a function of the redshift parametrized by two different relations: eta(z) = 1 +eta(0)z, and eta(z) = 1 + eta(0)z/(1 + z), where eta(0) is a constant parameter quantifying the possible departure from the strict validity of the DD relation. In order to determine the probability density function (PDF) of eta(0), we considered the angular diameter distances from galaxy clusters recently studied by two different groups by assuming elliptical and spherical isothermal beta models and spherical non-isothermal beta model. The strict validity of the DD relation will occur only if the maximum value of eta(0) PDF is centered on eta(0) = 0. Results. For both approaches we find that the elliptical beta model agrees with the distance-duality relation, whereas the non-isothermal spherical description is, in the best scenario, only marginally compatible. We find that the two-light curve fitters (SALT2 and MLCS2K2) present a statistically significant conflict, and a joint analysis involving the different approaches suggests that clusters are endowed with an elliptical geometry as previously assumed. Conclusions. The statistical analysis presented here provides new evidence that the true geometry of clusters is elliptical. In principle, it is remarkable that a local property such as the geometry of galaxy clusters might be constrained by a global argument like the one provided by the cosmological distance-duality relation.

Regional and temporal variability of sinking organic matter in the subtropical northeast Atlantic Ocean: a biomarker diagnosis

Sinking particles through the pelagic ocean have been traditionally considered the most important vehicle by which the biological pump sequesters carbon in the ocean interior. Nevertheless, regional scale variability in particle flux is a major outstanding issue in oceanography. 5 Here, we have studied the regional and temporal variability of total particulate organic matter fluxes, as well as chloropigment and total hydrolyzed amino acid (THAA) compositions and fluxes in the Canary Current region, between 20–30 N, during two contrasting periods: August 2006, characterized by warm and stratified waters, but also intense winds which enhanced eddy development south of the Canary Islands, 10 and February 2007, characterized by colder waters, less stratification and higher productivity. We found that the eddy-field generated south of the Canary Islands enhanced by >2 times particulate organic carbon (POC) export with respect to stations (FF; farfield) outside the eddy-field influence. We also observed flux increases of one order of magnitude in chloropigment and 70% in THAA in the eddy-field relative to FF stations. 15 Principal Components Analysis (PCA) was performed to assess changes in particulate organic matter composition between stations. At eddy-field stations, higher chlorophyll enrichment reflected “fresher” material, while at FF stations a higher proportion of pheophytin indicated greater degradation due to microbes and microzooplankton. PCA also suggests that phytoplankton community structure, particularly the dominance of 20 diatoms versus carbonate-rich plankton, is the major factor influencing the POC export within the eddy field. In February, POC export fluxes were the highest ever reported for this area, reaching values of 15 mmolCm−2 d−1 at 200m depth. Compositional changes in pigments and THAA indicate that the source of sinking particles varies zonally and meridionally and suggest that sinking particles were more degraded at 25 near-coastal stations relative to open ocean stations.

Regional and temporal variability of sinking organic matter in the subtropical northeast Atlantic Ocean: A biomarker diagnosis

[EN] Sinking particles through the pelagic ocean have been traditionally considered the most important vehicle by which the biological pump sequesters carbon in the ocean interior. Nevertheless, regional scale variability in particle flux is a major outstanding issue in oceanography. Here, we have studied the regional and temporal variability of total particulate organic matter fluxes, as well as chloropigment and total hydrolyzed amino acid (THAA) compositions and fluxes in the Canary Current region, between 20?30_ N, during two contrasting periods: August 2006, characterized by warm and stratified waters, but also intense winds which enhanced eddy development south of the Canary Islands, and February 2007, characterized by colder waters, less stratification and higher productivity. We found that the eddyfield generated south of the Canary Islands enhanced by >2 times particulate organic carbon (POC) export with respect to stations (FF; far-field) outside the eddy-field influence. We also observed flux increases of one order of magnitude in chloropigment and 2 times in THAA in the eddy-field relative to FF stations. Principal Components Analysis (PCA) was performed to assess changes in particulate organic matter composition between stations. At eddy-field stations, higher chlorophyll enrichment reflected ?fresher? material, while at FF stations a higher proportion of pheophytin indicated greater degradation due to microbes and microzooplankton. PCA also suggests that phytoplankton community structure, particularly the dominance of diatoms versus carbonate-rich plankton, is the major factor influencing the POC export within the eddy field. In February, POC export POC export within the eddy field. In February, POC export fluxes were the highest ever reported for this area, reaching values of _15 mmolCm?2 d?1 at 200m depth. Compositional changes in pigments and THAA indicate that the source of sinking particles varies zonally and meridionally and suggest that sinking particles were more degraded at near-coastal stations relative to open ocean stations.

Procedimenti di ottimizzazione di sistemi smorzati

La presente tesi riguarda lo studio di procedimenti di ottimizzazione di sistemi smorzati. In particolare, i sistemi studiati sono strutture shear-type soggette ad azioni di tipo sismico impresse alla base. Per effettuare l’ottimizzazione dei sistemi in oggetto si agisce sulle rigidezze di piano e sui coefficienti di smorzamento effettuando una ridistribuzione delle quantità suddette nei piani della struttura. È interessante effettuare l’ottimizzazione di sistemi smorzati nell’ottica della progettazione antisismica, in modo da ridurre la deformata della struttura e, conseguentemente, anche le sollecitazioni che agiscono su di essa. Il lavoro consta di sei capitoli nei quali vengono affrontate tre procedure numerico-analitiche per effettuare l’ottimizzazione di sistemi shear-type. Nel primo capitolo si studia l’ottimizzazione di sistemi shear-type agendo su funzioni di trasferimento opportunamente vincolate. In particolare, le variabili di progetto sono le rigidezze di piano, mentre i coefficienti di smorzamento e le masse di piano risultano quantità note e costanti durante tutto il procedimento di calcolo iterativo; per effettuare il controllo dinamico della struttura si cerca di ottenere una deformata pressoché rettilinea. Tale condizione viene raggiunta ponendo le ampiezze delle funzioni di trasferimento degli spostamenti di interpiano pari all’ampiezza della funzione di trasferimento del primo piano. Al termine della procedura si ottiene una ridistribuzione della rigidezza complessiva nei vari piani della struttura. In particolare, si evince un aumento della rigidezza nei piani più bassi che risultano essere quelli più sollecitati da una azione impressa alla base e, conseguentemente, si assiste ad una progressiva riduzione della variabile di progetto nei piani più alti. L’applicazione numerica di tale procedura viene effettuata nel secondo capitolo mediante l’ausilio di un programma di calcolo in linguaggio Matlab. In particolare, si effettua lo studio di sistemi a tre e a cinque gradi di libertà. La seconda procedura numerico-analitica viene presentata nel terzo capitolo. Essa riguarda l’ottimizzazione di sistemi smorzati agendo simultaneamente sulla rigidezza e sullo smorzamento e consta di due fasi. La prima fase ricerca il progetto ottimale della struttura per uno specifico valore della rigidezza complessiva e dello smorzamento totale, mentre la seconda fase esamina una serie di progetti ottimali in funzione di diversi valori della rigidezza e dello smorzamento totale. Nella prima fase, per ottenere il controllo dinamico della struttura, viene minimizzata la somma degli scarti quadratici medi degli spostamenti di interpiano. Le variabili di progetto, aggiornate dopo ogni iterazione, sono le rigidezze di piano ed i coefficienti di smorzamento. Si pone, inoltre, un vincolo sulla quantità totale di rigidezza e di smorzamento, e i valori delle rigidezze e dei coefficienti di smorzamento di ogni piano non devono superare un limite superiore posto all’inizio della procedura. Anche in questo caso viene effettuata una ridistribuzione delle rigidezze e dei coefficienti di smorzamento nei vari piani della struttura fino ad ottenere la minimizzazione della funzione obiettivo. La prima fase riduce la deformata della struttura minimizzando la somma degli scarti quadrarici medi degli spostamenti di interpiano, ma comporta un aumento dello scarto quadratico medio dell’accelerazione assoluta dell’ultimo piano. Per mantenere quest’ultima quantità entro limiti accettabili, si passa alla seconda fase in cui si effettua una riduzione dell’accelerazione attraverso l’aumento della quantità totale di smorzamento. La procedura di ottimizzazione di sistemi smorzati agendo simultaneamente sulla rigidezza e sullo smorzamento viene applicata numericamente, mediante l’utilizzo di un programma di calcolo in linguaggio Matlab, nel capitolo quattro. La procedura viene applicata a sistemi a due e a cinque gradi di libertà. L’ultima parte della tesi ha come oggetto la generalizzazione della procedura che viene applicata per un sistema dotato di isolatori alla base. Tale parte della tesi è riportata nel quinto capitolo. Per isolamento sismico di un edificio (sistema di controllo passivo) si intende l’inserimento tra la struttura e le sue fondazioni di opportuni dispositivi molto flessibili orizzontalmente, anche se rigidi in direzione verticale. Tali dispositivi consentono di ridurre la trasmissione del moto del suolo alla struttura in elevazione disaccoppiando il moto della sovrastruttura da quello del terreno. L’inserimento degli isolatori consente di ottenere un aumento del periodo proprio di vibrare della struttura per allontanarlo dalla zona dello spettro di risposta con maggiori accelerazioni. La principale peculiarità dell’isolamento alla base è la possibilità di eliminare completamente, o quantomeno ridurre sensibilmente, i danni a tutte le parti strutturali e non strutturali degli edifici. Quest’ultimo aspetto è importantissimo per gli edifici che devono rimanere operativi dopo un violento terremoto, quali ospedali e i centri operativi per la gestione delle emergenze. Nelle strutture isolate si osserva una sostanziale riduzione degli spostamenti di interpiano e delle accelerazioni relative. La procedura di ottimizzazione viene modificata considerando l’introduzione di isolatori alla base di tipo LRB. Essi sono costituiti da strati in elastomero (aventi la funzione di dissipare, disaccoppiare il moto e mantenere spostamenti accettabili) alternati a lamine in acciaio (aventi la funzione di mantenere una buona resistenza allo schiacciamento) che ne rendono trascurabile la deformabilità in direzione verticale. Gli strati in elastomero manifestano una bassa rigidezza nei confronti degli spostamenti orizzontali. La procedura di ottimizzazione viene applicata ad un telaio shear-type ad N gradi di libertà con smorzatori viscosi aggiunti. Con l’introduzione dell’isolatore alla base si passa da un sistema ad N gradi di libertà ad un sistema a N+1 gradi di libertà, in quanto l’isolatore viene modellato alla stregua di un piano della struttura considerando una rigidezza e uno smorzamento equivalente dell’isolatore. Nel caso di sistema sheat-type isolato alla base, poiché l’isolatore agisce sia sugli spostamenti di interpiano, sia sulle accelerazioni trasmesse alla struttura, si considera una nuova funzione obiettivo che minimizza la somma incrementata degli scarti quadratici medi degli spostamenti di interpiano e delle accelerazioni. Le quantità di progetto sono i coefficienti di smorzamento e le rigidezze di piano della sovrastruttura. Al termine della procedura si otterrà una nuova ridistribuzione delle variabili di progetto nei piani della struttura. In tal caso, però, la sovrastruttura risulterà molto meno sollecitata in quanto tutte le deformazioni vengono assorbite dal sistema di isolamento. Infine, viene effettuato un controllo sull’entità dello spostamento alla base dell’isolatore perché potrebbe raggiungere valori troppo elevati. Infatti, la normativa indica come valore limite dello spostamento alla base 25cm; valori più elevati dello spostamento creano dei problemi soprattutto per la realizzazione di adeguati giunti sismici. La procedura di ottimizzazione di sistemi isolati alla base viene applicata numericamente mediante l’utilizzo di un programma di calcolo in linguaggio Matlab nel sesto capitolo. La procedura viene applicata a sistemi a tre e a cinque gradi di libertà. Inoltre si effettua il controllo degli spostamenti alla base sollecitando la struttura con il sisma di El Centro e il sisma di Northridge. I risultati hanno mostrato che la procedura di calcolo è efficace e inoltre gli spostamenti alla base sono contenuti entro il limite posto dalla normativa. Giova rilevare che il sistema di isolamento riduce sensibilmente le grandezze che interessano la sovrastruttura, la quale si comporta come un corpo rigido al di sopra dell’isolatore. In futuro si potrà studiare il comportamento di strutture isolate considerando diverse tipologie di isolatori alla base e non solo dispositivi elastomerici. Si potrà, inoltre, modellare l’isolatore alla base con un modello isteretico bilineare ed effettuare un confronto con i risultati già ottenuti per il modello lineare.

Effetti del chitosano su composti polifenolici in colture cellulari di vite: aspetti molecolari e metabolici

Polyphenols, including flavonoids and stilbenes, are an essential part of human diet and constitute one of the most abundant and ubiquitous group of plant secondary metabolites. The level of these compounds is inducible by stress or fungal attack, so attempts are being made to identify likely biotic and abiotic elicitors and to better understand the underlying mechanism. Resveratrol (3,5,4’-trihydroxystilbene), which belongs to the stilbene family, is a naturally occurring polyphenol, found in several fruits, vegetables and beverages including red wine. It is one of the most important plant polyphenols with proved benefic activity on animal health. In the last two decades, the potential protective effects of resveratrol against cardiovascular and neurodegenerative diseases, as well as the chemopreventive properties against cancer, have been largely investigated. The most important source of polyphenols and in particular resveratrol for human diet is grape (Vitis vinifera). Since stilbenes and flavonoids play a very important role in plant defence responses and enviromental interactions, and their effects on human health seem promising, the aim of the research of this Thesis was to study at different levels the activation and the regulation of their biosynthetic pathways after chitosan treatment. Moreover, the polyphenol production in grape cells and the optimisation of cultural conditions bioreactor scale-up, were also investigated. Cell suspensions were obtained from cv. Barbera (Vitis vinifera L.) petioles and were treated with a biotic elicitor, chitosan (50 μg/mL, dissolved in acetic acid) to promote phenylpropanoid metabolism. Chitosan is a D-glucosamine polymer from fungi cell wall and therefore mimes fungal pathogen attack. Liquid cultures have been monitored for 15 days, measuring cell number, cell viability, pH and grams of fresh weight. The endogenous and released amounts of 7 stilbenes (trans and cis isomers of resveratrol, piceid and resveratroloside, and piceatannol), gallic acid, 6 hydroxycinnamic acids (trans-cinnamic, p-coumaric, caffeic, ferulic, sinapic and chlorogenic acids), 5 catechines (catechin, epicatechin, epigallocatechin-gallate (EGCG), epigallocatechin and epicatechin-gallate) and other 5 flavonoids (chalcon, naringenin, kaempferol, quercetin and rutin) in cells and cultural medium, were measured by HPLC-DAD analysis and total anthocyanins were quantified by spectrophotometric analysis. Chitosan was effective in stimulating trans-resveratrol endogenous accumulation with a sharp peak at day 4 (exceeding acetic acid and water controls by 36% and 63%, respectively), while it did not influence the production of the cis-isomer. Compared to both water and acetic acid controls, chitosan decreased the release of both trans- and cis-resveratrol respect to controls. No effect was shown on the accumulation of single resveratrol mono-glucoside isomers, but considering their total amount, normalized for the relative water control, it was possible to evidence an increase in both accumulation and release of those compounds, in chitosan-treated cells, throughout the culture period and particularly during the second week. Many of the analysed flavonoids and hydroxycinnamic acids were not present or detectable in trace amounts. Catechin, epicatechin and epigallocatechin-gallate (EGCG) were detectable both inside the cells and in the culture media, but chitosan did not affect their amounts. On the contrary, total anthocyanins have been stimulated by chitosan and their level, from day 4 to 14, was about 2-fold higher than in both controls, confirming macroscopic observations that treated suspensions showed an intense brown-red color, from day 3 onwards. These elicitation results suggest that chitosan selectively up-regulates specific biosynthetic pathways, without modifying the general accumulation pattern of other flavonoids. Proteins have been extracted from cells at day 4 of culture (corresponding to the production peak of trans-resveratrol) and separated by bidimensional electrophoresis. The 73 proteins that showed a consistently changed amount between untreated, chitosan and acetic acid (chitosan solvent) treated cells, have been identified by mass spectrometry. Chitosan induced an increase in stilbene synthase (STS, the resveratrol biosynthetic enzyme), chalcone-flavanone isomerase (CHI, that switches the pathway from chalcones to flavones and anthocyanins), pathogenesis-related proteins 10 (PRs10, a large family of defence proteins), and a decrease in many proteins belonging to primary metabolisms. A train of six distinct spots of STS encoded by the same gene and increased by chitosan, was detected on the 2-D gels, and related to the different phosphorylation degree of STS spots. Northern blot analyses have been performed on RNA extracted from cells treated with chitosan and relative controls, using probes for STS, PAL (phenylalanine ammonia lyase, the first enzyme of the biosynthetic pathway), CHS (chalcone synthase, that shares with STS the same precursors), CHI and PR-10. The up-regulation of PAL, CHS and CHI transcript expression levels correlated with the accumulation of anthocyanins. The strong increase of different molecular weight PR-10 mRNAs, correlated with the 11 PR-10 protein spots identified in proteomic analyses. The sudden decrease in trans-resveratrol endogenous accumulation after day 4 of culture, could be simply explained by the diminished resveratrol biosynthetic activity due to the lower amount of biosynthetic enzymes. This might be indirectly demonstrated by northern blot expression analyses, that showed lower levels of phenylalanine ammonia lyase (PAL) and stilbene synthase (STS) mRNAs starting from day 4. Other possible explanations could be a resveratrol oxidation process and/or the formation of other different mono-, di-glucosides and resveratrol oligomers such as viniferins. Immunolocalisation experiments performed on grape protoplasts and the subsequent analyses by confocal microscope, showed that STS, and therefore the resveratrol synthetic site, is mostly associated to intracellular membranes close to the cytosolic side of plasma membrane and in a smaller amount is localized in the cytosol. STS seemed not to be present inside vacuole and nucleus. There were no differences in the STS intracellular localisation between the different treatments. Since it was shown that stilbenes are largely released in the culture medium and that STS is a soluble protein, a possible interaction of STS with a plasma membrane transporter responsible for the extrusion of stilbenes in the culture medium, might be hypothesized. Proteomic analyses performed on subcellular fractions identified in the microsomial fraction 5 proteins taking part in channel complexes or associated with channels, that significantly changed their amount after chitosan treatment. In soluble and membrane fractions respectively 3 and 4 STS and 6 and 3 PR-10 have been identified. Proteomic results obtained from subcellular fractions substantially confirmed previous result obtained from total cell protein extracts and added more information about protein localisation and co-localisation. The interesting results obtained on Barbera cell cultures with the aim to increase polyphenol (especially stilbenes) production, have encouraged scale up tests in 1 litre bioreactors. The first trial fermentation was performed in parallel with a normal time-course in 20 mL flasks, showing that the scale-up (bigger volume and different conditions) process influenced in a very relevant way stilbenes production. In order to optimise culture parameters such as medium sucrose amount, fermentation length and inoculum cell concentration, few other fermentations were performed. Chitosan treatments were also performed. The modification of each parameter brought relevant variations in stilbenes and catechins levels, so that the production of a certain compound (or class of compounds) could be hypothetically promoted by modulating one or more culture parameters. For example the catechin yield could be improved by increasing sucrose content and the time of fermentation. The best results in stilbene yield were obtained in a 800 mL fermentation inoculated with 10.8 grams of cells and supplemented with chitosan. The culture was fed with MS medium added with 30 g/L sucrose, 25 μg/mL rifampicin and 50 μg/mL of chitosan, and was maintained at 24°C, stirred by marine impeller at 100 rpm and supplied of air at 0.16 L/min rate. Resveratroloside was the stilbene present in the larger amount, 3-5 times more than resveratrol. Because resveratrol glucosides are similarly active and more stable than free resveratrol, their production using a bioreactor could be a great advantage in an hypothetical industrial process. In my bioreactor tests, stilbenes were mainly released in the culture medium (60-80% of the total) and this fact could be another advantage for industrial applications, because it allows recovering the products directly from the culture medium without stopping the fermentation and/or killing the cells. In my best cultural conditions, it was possible to obtain 3.95 mg/L of stilbenes at day 4 (maximum resveratrol accumulation) and 5.13 mg/L at day 14 (maximum resveratroloside production). In conclusion, chitosan effect in inducing Vitis vinifera defense mechanisms can be related to its ability to increase the intracellular content of a large spectrum of antioxidants, and in particular of resveratrol, its derivates and anthocyanins. Its effect can be observed at transcriptional, proteomic (variation of soluble and membrane protein amounts) and metabolic (polyphenols production) level. The chitosan ability to elicit specific plant matabolisms can be useful to produce large quantities of antioxidant compounds from cell culture in bioreactor.

Studio e sviluppo di un motore a 2 tempi diesel veloce di nuova concezione per applicazioni automobilistiche

The Department of Mechanical and Civil Engineering (DIMeC) of the University of Modena and Reggio Emilia is developing a new type of small capacity HSDI 2-Stroke Diesel engine (called HSD2), featuring a specifically designed combustion system, aimed to reduce weight, size and manufacturing costs, and improving pollutant emissions at partial load. The present work is focused on the analysis of the combustion and the scavenging process, investigated by means of a version of the KIVA-3V code customized by the University of Chalmers and modified by DIMeC. The customization of the KIVA-3V code includes a detailed combustion chemistry approach, coupled with a comprehensive oxidation mechanism for diesel oil surrogate and the modeling of turbulence/chemistry interaction through the PaSR (Partially Stirred Reactor) model. A four stroke automobile Diesel engine featuring a very close bore size is taken as a reference, for both the numerical models calibration and for a comparison with the 2-Stroke engine. Analysis is carried out trough a comparison between HSD2 and FIAT 1300 MultiJet in several operating conditions, at full and partial load. Such a comparison clearly demonstrates the effectiveness of the two stroke concept in terms of emissions reduction and high power density. However, HSD2 is still a virtual engine, and experimental results are needed to assume the reliability of numerical results.

Technical innovations for the diagnosis and the rehabilitation of motor and perceptive impairments of the child with Cerebral Palsy

The treatment of the Cerebral Palsy (CP) is considered as the “core problem” for the whole field of the pediatric rehabilitation. The reason why this pathology has such a primary role, can be ascribed to two main aspects. First of all CP is the form of disability most frequent in childhood (one new case per 500 birth alive, (1)), secondarily the functional recovery of the “spastic” child is, historically, the clinical field in which the majority of the therapeutic methods and techniques (physiotherapy, orthotic, pharmacologic, orthopedic-surgical, neurosurgical) were first applied and tested. The currently accepted definition of CP – Group of disorders of the development of movement and posture causing activity limitation (2) – is the result of a recent update by the World Health Organization to the language of the International Classification of Functioning Disability and Health, from the original proposal of Ingram – A persistent but not unchangeable disorder of posture and movement – dated 1955 (3). This definition considers CP as a permanent ailment, i.e. a “fixed” condition, that however can be modified both functionally and structurally by means of child spontaneous evolution and treatments carried out during childhood. The lesion that causes the palsy, happens in a structurally immature brain in the pre-, peri- or post-birth period (but only during the firsts months of life). The most frequent causes of CP are: prematurity, insufficient cerebral perfusion, arterial haemorrhage, venous infarction, hypoxia caused by various origin (for example from the ingestion of amniotic liquid), malnutrition, infection and maternal or fetal poisoning. In addition to these causes, traumas and malformations have to be included. The lesion, whether focused or spread over the nervous system, impairs the whole functioning of the Central Nervous System (CNS). As a consequence, they affect the construction of the adaptive functions (4), first of all posture control, locomotion and manipulation. The palsy itself does not vary over time, however it assumes an unavoidable “evolutionary” feature when during growth the child is requested to meet new and different needs through the construction of new and different functions. It is essential to consider that clinically CP is not only a direct expression of structural impairment, that is of etiology, pathogenesis and lesion timing, but it is mainly the manifestation of the path followed by the CNS to “re”-construct the adaptive functions “despite” the presence of the damage. “Palsy” is “the form of the function that is implemented by an individual whose CNS has been damaged in order to satisfy the demands coming from the environment” (4). Therefore it is only possible to establish general relations between lesion site, nature and size, and palsy and recovery processes. It is quite common to observe that children with very similar neuroimaging can have very different clinical manifestations of CP and, on the other hand, children with very similar motor behaviors can have completely different lesion histories. A very clear example of this is represented by hemiplegic forms, which show bilateral hemispheric lesions in a high percentage of cases. The first section of this thesis is aimed at guiding the interpretation of CP. First of all the issue of the detection of the palsy is treated from historical viewpoint. Consequently, an extended analysis of the current definition of CP, as internationally accepted, is provided. The definition is then outlined in terms of a space dimension and then of a time dimension, hence it is highlighted where this definition is unacceptably lacking. The last part of the first section further stresses the importance of shifting from the traditional concept of CP as a palsy of development (defect analysis) towards the notion of development of palsy, i.e., as the product of the relationship that the individual however tries to dynamically build with the surrounding environment (resource semeiotics) starting and growing from a different availability of resources, needs, dreams, rights and duties (4). In the scientific and clinic community no common classification system of CP has so far been universally accepted. Besides, no standard operative method or technique have been acknowledged to effectively assess the different disabilities and impairments exhibited by children with CP. CP is still “an artificial concept, comprising several causes and clinical syndromes that have been grouped together for a convenience of management” (5). The lack of standard and common protocols able to effectively diagnose the palsy, and as a consequence to establish specific treatments and prognosis, is mainly because of the difficulty to elevate this field to a level based on scientific evidence. A solution aimed at overcoming the current incomplete treatment of CP children is represented by the clinical systematic adoption of objective tools able to measure motor defects and movement impairments. A widespread application of reliable instruments and techniques able to objectively evaluate both the form of the palsy (diagnosis) and the efficacy of the treatments provided (prognosis), constitutes a valuable method able to validate care protocols, establish the efficacy of classification systems and assess the validity of definitions. Since the ‘80s, instruments specifically oriented to the analysis of the human movement have been advantageously designed and applied in the context of CP with the aim of measuring motor deficits and, especially, gait deviations. The gait analysis (GA) technique has been increasingly used over the years to assess, analyze, classify, and support the process of clinical decisions making, allowing for a complete investigation of gait with an increased temporal and spatial resolution. GA has provided a basis for improving the outcome of surgical and nonsurgical treatments and for introducing a new modus operandi in the identification of defects and functional adaptations to the musculoskeletal disorders. Historically, the first laboratories set up for gait analysis developed their own protocol (set of procedures for data collection and for data reduction) independently, according to performances of the technologies available at that time. In particular, the stereophotogrammetric systems mainly based on optoelectronic technology, soon became a gold-standard for motion analysis. They have been successfully applied especially for scientific purposes. Nowadays the optoelectronic systems have significantly improved their performances in term of spatial and temporal resolution, however many laboratories continue to use the protocols designed on the technology available in the ‘70s and now out-of-date. Furthermore, these protocols are not coherent both for the biomechanical models and for the adopted collection procedures. In spite of these differences, GA data are shared, exchanged and interpreted irrespectively to the adopted protocol without a full awareness to what extent these protocols are compatible and comparable with each other. Following the extraordinary advances in computer science and electronics, new systems for GA no longer based on optoelectronic technology, are now becoming available. They are the Inertial and Magnetic Measurement Systems (IMMSs), based on miniature MEMS (Microelectromechanical systems) inertial sensor technology. These systems are cost effective, wearable and fully portable motion analysis systems, these features gives IMMSs the potential to be used both outside specialized laboratories and to consecutive collect series of tens of gait cycles. The recognition and selection of the most representative gait cycle is then easier and more reliable especially in CP children, considering their relevant gait cycle variability. The second section of this thesis is focused on GA. In particular, it is firstly aimed at examining the differences among five most representative GA protocols in order to assess the state of the art with respect to the inter-protocol variability. The design of a new protocol is then proposed and presented with the aim of achieving gait analysis on CP children by means of IMMS. The protocol, named ‘Outwalk’, contains original and innovative solutions oriented at obtaining joint kinematic with calibration procedures extremely comfortable for the patients. The results of a first in-vivo validation of Outwalk on healthy subjects are then provided. In particular, this study was carried out by comparing Outwalk used in combination with an IMMS with respect to a reference protocol and an optoelectronic system. In order to set a more accurate and precise comparison of the systems and the protocols, ad hoc methods were designed and an original formulation of the statistical parameter coefficient of multiple correlation was developed and effectively applied. On the basis of the experimental design proposed for the validation on healthy subjects, a first assessment of Outwalk, together with an IMMS, was also carried out on CP children. The third section of this thesis is dedicated to the treatment of walking in CP children. Commonly prescribed treatments in addressing gait abnormalities in CP children include physical therapy, surgery (orthopedic and rhizotomy), and orthoses. The orthotic approach is conservative, being reversible, and widespread in many therapeutic regimes. Orthoses are used to improve the gait of children with CP, by preventing deformities, controlling joint position, and offering an effective lever for the ankle joint. Orthoses are prescribed for the additional aims of increasing walking speed, improving stability, preventing stumbling, and decreasing muscular fatigue. The ankle-foot orthosis (AFO), with a rigid ankle, are primarily designed to prevent equinus and other foot deformities with a positive effect also on more proximal joints. However, AFOs prevent the natural excursion of the tibio-tarsic joint during the second rocker, hence hampering the natural leaning progression of the whole body under the effect of the inertia (6). A new modular (submalleolar) astragalus-calcanear orthosis, named OMAC, has recently been proposed with the intention of substituting the prescription of AFOs in those CP children exhibiting a flat and valgus-pronated foot. The aim of this section is thus to present the mechanical and technical features of the OMAC by means of an accurate description of the device. In particular, the integral document of the deposited Italian patent, is provided. A preliminary validation of OMAC with respect to AFO is also reported as resulted from an experimental campaign on diplegic CP children, during a three month period, aimed at quantitatively assessing the benefit provided by the two orthoses on walking and at qualitatively evaluating the changes in the quality of life and motor abilities. As already stated, CP is universally considered as a persistent but not unchangeable disorder of posture and movement. Conversely to this definition, some clinicians (4) have recently pointed out that movement disorders may be primarily caused by the presence of perceptive disorders, where perception is not merely the acquisition of sensory information, but an active process aimed at guiding the execution of movements through the integration of sensory information properly representing the state of one’s body and of the environment. Children with perceptive impairments show an overall fear of moving and the onset of strongly unnatural walking schemes directly caused by the presence of perceptive system disorders. The fourth section of the thesis thus deals with accurately defining the perceptive impairment exhibited by diplegic CP children. A detailed description of the clinical signs revealing the presence of the perceptive impairment, and a classification scheme of the clinical aspects of perceptual disorders is provided. In the end, a functional reaching test is proposed as an instrumental test able to disclosure the perceptive impairment. References 1. Prevalence and characteristics of children with cerebral palsy in Europe. Dev Med Child Neurol. 2002 Set;44(9):633-640. 2. Bax M, Goldstein M, Rosenbaum P, Leviton A, Paneth N, Dan B, et al. Proposed definition and classification of cerebral palsy, April 2005. Dev Med Child Neurol. 2005 Ago;47(8):571-576. 3. Ingram TT. A study of cerebral palsy in the childhood population of Edinburgh. Arch. Dis. Child. 1955 Apr;30(150):85-98. 4. Ferrari A, Cioni G. The spastic forms of cerebral palsy : a guide to the assessment of adaptive functions. Milan: Springer; 2009. 5. Olney SJ, Wright MJ. Cerebral Palsy. Campbell S et al. Physical Therapy for Children. 2nd Ed. Philadelphia: Saunders. 2000;:533-570. 6. Desloovere K, Molenaers G, Van Gestel L, Huenaerts C, Van Campenhout A, Callewaert B, et al. How can push-off be preserved during use of an ankle foot orthosis in children with hemiplegia? A prospective controlled study. Gait Posture. 2006 Ott;24(2):142-151.