Cassava starch biodegradable films: Influence of glycerol and clay nanoparticles content on tensile and barrier properties and glass transition temperature

In this this study, glycerol content and its incorporation method on tensile and barrier properties of biodegradable films (BF) based on cassava starch were analyzed. ANOVA showed that the glycerol incorporation method did not influence the results (P > 0.05), however the glycerol content influenced significantly the tensile and barrier properties of the films (P < 0.05). Films prepared with lower glycerol content presented better tensile and barrier properties than films with higher content. Films were then prepared with addition of clay nanoparticles and their tensile and barrier properties and glass transition temperature were measured. ANOVA indicated that both glycerol and clay nanoparticles influenced significantly the tensile and barrier properties (P < 0.05), diminishing film permeability when clay nanoparticles were present, while the glass transition temperature was not influenced (P > 0.05). (C) 2011 Elsevier Ltd. All rights reserved.

Comparison between innominate osteotomy and arthrodistraction as a primary treatment for Legg-Calve-Perthes disease: a prospective controlled trial

Legg-Calv,-Perthes (LCP) disease is currently managed by mechanical containment of the femoral head in the hip socket. As evidence suggests that hip distraction may offer a new treatment strategy, we used arthrodistraction as a primary treatment for active forms of LCP disease and prospectively compared the results with the Salter innominate osteotomy. A total of 54 children, six years or older of both genders with severe forms of LCP disease in the stages of necrosis or revascularisation, were enrolled. Patients were submitted to either Salter innominate osteotomy (n = 28) or hip arthrodistraction (n = 26). Final radiographs were used to evaluate the Mose index, Wiberg angle, extrusion index and the Stulberg et al. classification. There were no significant differences in gender, age, lateral pillar classification and average follow-up time between the two groups. The osteotomy group progressed without major complications, but children in the joint distraction group experienced episodes of pin tract pain and infection, leading to the early removal of the external device in one case. Two patients developed joint stiffness, treated by physiotherapy or manipulation, and one child developed subluxation of the femoral head. The average time in distraction was 4.44 months (2.53-7.23 months). In the final evaluation the osteotomy group showed better containment of the femoral head. The Mose index and the Stulberg et al. classification were statistically similar between the two groups. Despite similar final radiological results, arthrodistraction was associated with a higher morbidity. Consequently, we do not recommend hip distraction as a primary treatment for the early stages of LCP disease.

Expression of VEGF and collagen using a latex biomembrane as bladder replacement in rabbits

Objective: To investigate the VEGF expression and collagen deposition using a latex biomembrane as bladder replacement in rabbits. Materials and Methods: After partial cystectomy, a patch of a non-vulcanized latex biomembrane (2 x 2 cm) was sewn to the bladder of rabbits with 5/0 monofilament polydioxanone sulfate sutures in a watertight manner. Groups of 5 animals were killed at 15, 45 and 90 days after surgery and the bladder was removed. Sections of 5 mu m were cut and stained with picrosirius-red in order to estimate the amount of extracellular matrix in the graft. To confirm the presence of VEGF in tissues, protein expression was determined by immunohistochemistry. Results: No death, urinary leakage or graft extrusion occurred in any group. All bladders showed a spherical shape. A progressive reduction in the amount of collagen occurred in the graft area and was negatively and linearly correlated with time (p < 0.001). VEGF expression was higher in grafted areas when compared to controls at 15 and 45 days after surgery and decreased with time (p < 0.001). Conclusion: The latex biomembrane as a matrix for partial bladder replacement in rabbits promotes temporary collagen deposition and stimulates the angiogenic process.

Left ventricular free wall impeding rupture in post-myocardial infarction period diagnosed by myocardial contrast echocardiography: Case report

Abstract Background: Left ventricular free wall rupture occurs in up to 10% of the in-hospital deaths following myocardial infarction. It is mainly associated with posterolateral myocardial infarction and its antemortem diagnosis is rarely made. Contrast echocardiography has been increasingly used for the evaluation of myocardial perfusion in patients with acute myocardial infarction, with important prognostic implications. In this case, we reported its use for the detection of a mechanical complication following myocardial infarction. Case presentation: A 50-year-old man with acute myocardial infarction in the lateral wall underwent myocardial contrast echocardiography for the evaluation of myocardial perfusion in the third day post-infarction. A perfusion defect was detected in lateral and inferior walls as well as the presence of contrast extrusion from the left ventricular cavity into the myocardium, forming a serpiginous duct extending from the endocardium to the epicardial region of the lateral wall, without communication with the pericardial space. Magnetic resonance imaging confirmed the diagnosis of impending rupture of the left ventricular free wall. While waiting for cardiac surgery, patient presented with cardiogenic shock and died. Anatomopathological findings were consistent with acute myocardial infarction in the lateral wall and a left ventricular free wall rupture at the infarct site. Conclusion: This case illustrates the early diagnosis of left ventricular free wall rupture by contrast echocardiography. Due to its ability to be performed at bedside this modality of imaging has the potential to identify this catastrophic condition in patients with acute myocardial infarction and help to treat these patients with emergent surgery.

Dentoskeletal changes induced by the Jasper jumper and the activator-headgear combination appliances followed by fixed orthodontic treatment

Introduction: The aim of this study was to compare the dentoskeletal changes of patients with Class II Division 1 malocclusion treated with either the Jasper jumper appliance or the activator-headgear combination, both associated with fixed appliances. Methods: The sample comprised 72 subjects with Class II Division 1 malocclusion divided into 3 groups: group 1 included 25 subjects treated with fixed appliances and the force modules of the Jasper jumper at an initial mean age of 12.72 years, group 2 included 25 subjects treated with the activator-headgear combination followed by fixed appliances at an initial mean age of 11.07 years, and group 3 included 22 untreated subjects at an initial mean age of 12.67 years. Initial cephalometric characteristics and dentoskeletal changes were compared with analysis of variance. Results: Both experimental groups had similar dentoskeletal changes: restrictive effect on the maxilla, clockwise mandibular rotation and a slight increase in anterior face height, retrusion of the maxillary incisors, distalization of the maxillary molars, protrusion of the mandibular incisors, extrusion of the mandibular molars, and significant improvements of the maxillomandibular relationship, overjet, overbite, and the molar relationship. Conclusions: The effects of the Jasper jumper and the activator-headgear combination followed by fixed orthodontic appliances were similar in Class II malocclusion treatment.

Push-out bond strengths of different dental cements used to cement glass fiber posts

Statement of problem: Since the introduction of glass fiber posts, irreversible vertical root fractures have become a rare occurrence; however, adhesive failure has become the primary failure mode. Purpose: The purpose of this study was to evaluate the push-out bond strength of glass fiber posts cemented with different luting agents on 3 segments of the root. Material and methods: Eighty human maxillary canines with similar root lengths were randomly divided into 8 groups (n=10) according to the cement assessed (Rely X luting, Luting and Lining, Ketac Cem, Rely X ARC, Biscem, Duo-link, Rely X U100, and Variolink II). After standardized post space preparation, the root dentin was pretreated for dualpolymerizing resin cements and untreated for the other cements. The mixed luting cement paste was inserted into post spaces with a spiral file and applied to the post surface that was seated into the canal. After 7 days, the teeth were sectioned perpendicular to their long axis into 1-mm-thick sections. The push-out test was performed at a speed of 0.5 mm/min until extrusion of the post occurred. The results were evaluated by 2-way ANOVA and the all pairwise multiple comparison procedures (Tukey test) (?=.05). Results: ANOVA showed that the type of interaction between cement and root location significantly influenced the push-out strength (P<.05). The highest push-out strength results with root location were obtained with Luting and Lining (S3) (19.5 ±4.9 MPa), Ketac Cem (S2) (18.6 ±5.5 MPa), and Luting and Lining (S1) (18.0 ±7.6 MPa). The lowest mean values were recorded with Variolink II (S1) (4.6 ±4.0 MPa), Variolink II (S2) (1.6 ±1.5 MPa), and Rely X ARC (S3) (0.9 ±1.1 MPa). Conclusions: Self-adhesive cements and glass ionomer cements showed significantly higher values compared to dual-polymerizing resin cements. In all root segments, dual-polymerizing resin cements provided significantly lower bond strength. Significant differences among root segments were found only for Duo-link cement.

Avaliação clínica e radiográfica do extravasamento da pasta de hidróxido de cálcio em dentes com lesões periapicais

It has been evaluated clinically and radiographically the effect of the extrusion of calcium hydroxide paste in teeth with periapical lesions. Twenty-five patients with teeth showing pulp necrosis and periapical lesions were submitted to endodontic treatment. Dressing calcium hydroxide paste, iodoform and propylene glycol were used. Clinically, the symptoms were evaluated after treatment and, radiographically, the reabsorption of extravasated paste and repair process of the periapical lesion. Only three patients had symptoms severe in the first 24 hours, the paste was completely reabsorbed within 15 days and no interference in the repair process.

Evaluation of apically extruded debris during endodontic retreatment

Introduction: A growing interest to preserve teeth into the mouth by patients resulted in the increasing number of endodontic retreatments, and when these happen, many different types of irritants are extruded through the foramen. Objective: This study analyzed in vitro the amount of debris extruded through the foramen using four instrumentation techniques during endodontic retreatment. Material and methods: Forty mesial-buccal roots of first molars were selected, instrumented with anatomical diameter up to size #30 ISO file and then obturated with gutta-percha and grossman sealer by lateral condensation. After, they were separated and randomly allocated into four groups with 10 teeth each for the endodontic retreatment procedure: G1 – conventional technique + solvent, G2 – conventional technique without solvent, G3 – ProTaper retreatment + solvent, G4 – ProTaper retreatment without solvent. In all groups, gutta-percha in the coronal portion was removed by using size 1-3 Gates Glidden drills. All teeth were irrigated with distilled water. The debris extruded through the foramen were collected and weighed by an analytical balance. Results: Group 4 had the lowest average for material extrusion through the foramen followed by groups 2, 3 and 1. When Tukey test for statistical analysis was applied, no significant difference among groups were found (p = 0.5664). Conclusion: We conclude that all instrumentation techniques used in this study produced debris which goes beyond the foramen.

Healing of a tooth with an overinstrumented apex, extensive transportation and periapical lesion using a 5 mm calcium hydroxide apical plug: an 8-year follow-up report

Besides the risk of filling material extrusion throughout the apex, a satisfactory apical seal can be difficult to achieve in canals with open apices or iatrogenic enlargements of the apical constriction. These situations pose a challenge to root canal filling. This paper describes the root canal filling of a maxillary right canine with an overinstrumented apex, complete loss of the apical stop, extensive canal transportation and apical periodontitis. A 5 mm calcium hydroxide apical plug was placed before root canal filling. The plug was made by soaking paper points with saline, dipping the points in calcium hydroxide powder and then applying it to the apex several times, until a consistent apical plug was obtained. The canal was then irrigated with saline in order to remove any residual calcium hydroxide from the root canal walls, dried with paper points and obturated with an inverted #80 gutta-percha cone and zinc oxide-eugenol based sealer by the lateral condensation technique. An 8-year radiographic follow-up showed formation of mineralized tissue sealing the apical foramen, apical remodeling and no signs of apical periodontitis.

Embryonic development and larval stages of Steindachneridion parahybae (Siluriformes: Pimelodidae): implications for the conservation and rearing of this endangered Neotropical species

Steindachneridion parahybae is a freshwater catfish endemic to the Paraíba do Sul River and is classified as an endangered Neotropical species. An increasing number of conservation biologists are incorporating morphological and physiological research data to help conservation managers in rescue these endangered species. This study investigated the embryonic and larval development of S. parahybae in captivity, with emphasis in major events during the ontogeny of S. parahybae. Broodstocks were artificially induced to reproduce, and the extrusion occurred 200-255 degree-hours after hormonal induction at 24°C. Larval ontogeny was evaluated every 10 minutes under microscopic/stereomicroscopic using fresh eggs samples. The main embryogenic development stages were identified: zygote, cleavage, including the morula, blastula, gastrula phase, organogenesis, and hatching. The extruded oocytes showed an average diameter of 1.10 ± 0.10 mm, and after fertilization and hydration of eggs, the average diameter of eggs increased to about 1.90 ± 0.60 mm, characterized by a large perivitelline space that persisted up to embryo development, the double chorion, and the poles (animal and vegetative). Cell division started about 2 minutes after fertilization (AF), resulting in 2, 4, 8 (4 x 2 arrangement of cells), 16 (4 x 4), 32 (4 x 8) and 64 (2 x 4 x 8) cells. Furthermore, the blastula and gastrula stages followed after these cells divisions. The closed blastopore occurred at 11 h 20 min AF; following the development, the organogenetic stages were identified and subdivided respectively in: early segmentation phase and late segmentation phase. In the early segmentation phase, there was the establishment of the embryonic axis, and it was possible to distinguish between the cephalic and caudal regions; somites, and the optic vesicles developed about 20 h AF. Total hatching occurred at 54 h AF, and the larvae average length was 4.30 ± 0.70 mm. Gradual yolk sac reduction was observed during the first two days of larval development. The first feeding occurred at the end of the second day. During the larval phase, cannibalism, heterogeneous larval growth and photophobia were also observed. This information will be important in improving the artificial reproduction protocols of S. parahybae in controlled breeding programs.

Avaliação da qualidade tecnológica de snacks obtidos por extrusão de grão integral de amaranto ou de farinha de amaranto desengordurada e suas misturas com fubá de milho

Apesar dos benefícios nutricionais e de seu potencial como alimento funcional, o amaranto ainda é um alimento pouco difundido no Brasil. Para o aumento do consumo desta matéria-prima, preconiza-se a sua incorporação na formulação de produtos convencionais, como os snacks extrudados que vêm sendo elaborados com a farinha de amaranto desengordurada. Pouco tem sido pesquisado sobre a extrusão direta do grão integral de amaranto, o que permitiria eliminar as etapas de moagem e de desengorduramento. Assim, a pesquisa avaliou e comparou a qualidade tecnológica de snacks obtidos por extrusão de grão integral de amaranto e de farinha de amaranto desengordurada, e suas misturas com 25 e 50% de fubá de milho. Verificou-se que o teor de lipídeos presentes no grão de amaranto (8%) prejudicou a expansão e a textura dos extrudados. Mesmo com a adição de fubá de milho, os snacks extrudados à base de grão integral de amaranto foram rejeitados sensorialmente (aceitação global entre 3 e 4, numa escala hedônica estruturada de nove pontos), pois apresentaram baixa expansão, textura dura, cor escura e forte sabor residual. Já os snacks extrudados obtidos com farinha de amaranto desengordurada isoladamente e em combinação com fubá de milho apresentaram maior expansão, textura crocante e cor mais clara, e, por isso, maior aceitabilidade (aceitação global > 5). Conclui-se, nas condições do experimento, que não foi possível obter snacks extrudados à base de grãos de amaranto de boa aceitação e que a produção de snacks extrudados à base de farinha de amaranto desengordurada proporciona produtos com melhor aceitação por parte do consumidor.

Cephalometric effects of the use of 10-hour Force Theory for Class II treatment

OBJECTIVE: This study aimed to evaluate the cephalometric effects promoted by the orthodontic treatment of Class II malocclusion patients with the use of the 10-Hour Force Theory, that consists in the use of fixed appliances with 8 hours a day using a cervical headgear appliance and 16 hours a day using Class II elastics, 8 hours on the first mandibular molar and 8 hours in the second mandibular molar. METHODS: Sample comprised 31 patients with mean initial age of 14.90 years, final mean age of 17.25 years and mean treatment time of 2.35 years. The lateral cephalograms in pre-treatment and post-treatment stages were evaluated. Evaluation of cephalometric changes between initial and final treatment phases was performed by paired t test. RESULTS: The cases treated with the 10-Hour Force Theory presented a slight restriction of anterior displacement of the maxilla, increase in the effective length of the mandible, significant improvement of the maxillomandibular relationship, significant increase in anterior lower face height, distal tipping of the maxillary premolar crowns, extrusion and distal tipping of the roots of maxillary molars, significant proclination and protrusion of mandibular incisors, significant extrusion and mesialization of mandibular molars, besides a significant correction of the molar relationship, overjet and overbite. CONCLUSION: The use of the 10-Hour Force Theory in treatment of Class II malocclusion provided satisfactory results.

Neoproterozoic evolution of the basement of the South-American platform

Neoproterozoic geologic and geotectonic processes were of utmost importance in forming and structuring the basement framework of the South-American platform. Two large domains with distinct evolutionary histories are identified with respect to the Neoproterozoic era: the northwest-west (Amazonian craton and surroundings) and the central-southeast (the extra-Amazonian domain). In the first domain, Neoproterozoic events occurred only locally and were of secondary significance, and the geologic events, processes, and structures of the pre-Neoproterozoic (and syn-Brasiliano) cratonic block were much more influential. In the second, the extra-Amazonian domain, the final evolution, structures and forms are assigned to events related to the development of a complex net of Neoproterozoic mobile belts. These in turn resulted in strong reworking of the older pre-Neoproterozoic basement. In this domain, four distinct structural provinces circumscribe or are separated by relatively small pre- Neoproterozoic cratonic nuclei, namely the Pampean, Tocantins, Borborema and Mantiqueira provinces. These extra-Amazonian provinces were formed by a complex framework of orogenic branching systems following a diversified post-Mesoproterozoic paleogeographic scenario. This scenario included many types of basement inliers as well as a diversified organization of accretionary and collisional orogens. The basement inliers date from the Archean toMesoproterozoic periods and are different in nature. The escape tectonics that operated during the final consolidation stages of the provinces were important to and responsible for the final forms currently observed. These latest events, which occurred from the Late Ediacaran to the Early Ordovician, present serious obstacles to paleogeographic reconstructions. Two groups of orogenic collage systems are identified. The older system from the Tonian (>850 Ma) period is of restricted occurrence and is not fully understood due to strong reworking subsequent to Tonian times. The second group of orogenies is more extensive and more important. Its development began with diachronic taphrogenic processes in the Early Cryogenian period (ca. 850e750 Ma) and preceded a complex scenario of continental, transitional and oceanic basins. Subsequent orogenies (post 800 Ma) were also created by diachronic processes that ended in the Early Ordovician. More than one orogeny (plate interaction) can be identified either in space or in time in every province. The orogenic processes were not necessarily synchronous in different parts of the orogenic system, even within the same province. This particular group of orogenic collage events is known as the “Brasiliano”. All of the structural provinces of the extra-Amazonian domain exhibit final events that are marked by extrusion processes, are represented by long lineaments, and are fundamental to unraveling the structural history of the Phanerozoic sedimentary basins.

Analisi sperimentale degli effetti di temperature elevate sul comportamento strutturale dei rinforzi in FRP

In questa tesi si è voluta porre l’attenzione sulla suscettibilità alle alte temperature delle resine che li compongono. Lo studio del comportamento alle alte temperature delle resine utilizzate per l’applicazione dei materiali compositi è risultato un campo di studio ancora non completamente sviluppato, nel quale c’è ancora necessità di ricerche per meglio chiarire alcuni aspetti del comportamento. L’analisi di questi materiali si sviluppa partendo dal contesto storico, e procedendo successivamente ad una accurata classificazione delle varie tipologie di materiali compositi soffermandosi sull’ utilizzo nel campo civile degli FRP (Fiber Reinforced Polymer) e mettendone in risalto le proprietà meccaniche. Considerata l’influenza che il comportamento delle resine riveste nel comportamento alle alte temperature dei materiali compositi si è, per questi elementi, eseguita una classificazione in base alle loro proprietà fisico-chimiche e ne sono state esaminate le principali proprietà meccaniche e termiche quali il modulo elastico, la tensione di rottura, la temperatura di transizione vetrosa e il fenomeno del creep. Sono state successivamente eseguite delle prove sperimentali, effettuate presso il Laboratorio Resistenza Materiali e presso il Laboratorio del Dipartimento di Chimica Applicata e Scienza dei Materiali, su dei provini confezionati con otto differenti resine epossidiche. Per valutarne il comportamento alle alte temperature, le indagini sperimentali hanno valutato dapprima le temperature di transizione vetrosa delle resine in questione e, in seguito, le loro caratteristiche meccaniche. Dalla correlazione dei dati rilevati si sono cercati possibili legami tra le caratteristiche meccaniche e le proprietà termiche delle resine. Si sono infine valutati gli aspetti dell’applicazione degli FRP che possano influire sul comportamento del materiale composito soggetto alle alte temperature valutando delle possibili precauzioni che possano essere considerate in fase progettuale.

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.