Effets préventifs et thérapeutiques des polyphénols dans un modèle in vitro et in vivo de maladie inflammatoire de l’intestin : caractérisation des polyphénols de la pelure de pomme et de la canneberge par spectrométrie de masse

La muqueuse intestinale est exposée à des agents oxydants provenant de l’ingestion d’aliments modifiés, de cellules immuno-inflammatoires et de la flore intestinale. Une diète élevée en fruits et légumes peut diminuer le stress oxydant (SOx) ainsi que l’inflammation via plusieurs mécanismes. Ces effets bénéfiques peuvent être attribuables à leur contenu élevé en polyphénols. La première étude de mon doctorat consistait à tester l’hypothèse que les polyphénols extraits de pelures de pomme (DAPP) pouvaient diminuer le stress oxydant et l'inflammation impliqués dans les maladies inflammatoires de l'intestin (MII). Nous avons caractérisé les polyphénols des DAPP par spectrométrie de masse (LC-MS) et examiné leur potentiel antioxydant et anti-inflammatoire au niveau des cellules intestinales. L’identification des structures chimiques des polyphénols a été effectuée par LC-MS. Le SOx a été induit par l’ajout du complexe fer/ascorbate (Fe/Asc, 200 µM/2 mM) et l’inflammation par la lipopolysaccharide (LPS, 200µg/mL) à des cellules intestinales Caco-2/15 pré-incubées avec les DAPP (250 µg/mL). L’effet du SOx est déterminé par le dosage du malondialdéhyde (MDA), de la composition des acides gras polyinsaturés et de l’activité des enzymes antioxydantes endogènes (SOD et GPx). L’impact des DAPP sur l’inflammation a été testé par l’analyse de l’expression des marqueurs inflammatoires: cyclooxygénase-2 (COX-2), le facteur de nécrose tumorale alpha (TNF-a et l’interleukine-6 (IL-6) et les facteurs de transcription NF-KB, Nrf-2 et PGC1α par immunobuvardage. Nos données ont montré que les flavonols et les flavan-3-ols constituent les composés polyphénoliques majoritaires des DAPP. L’ajout de Fer2+/Asc a provoqué une augmentation de la peroxidation lipidique comparativement aux cellules contrôles, un appauvrissement des acides gras polyinsaturés n-3 et n-6, et une modulation des enzymes antioxydantes, se traduisant par une augmentation de l’activité de la SOD et une diminution de la GPx. En contrepartie, les DAPP ont exhibé leur potentiel à corriger la plupart des perturbations, y compris l’expression protéique anormalement élevée du COX-2 et la production de la prostaglandine E2 (PGE2), ainsi que l’inflammation telle que réflétée par les facteurs NF-κB, TNF-α et IL-6. Par ailleurs, les mécanismes sous-jacents à ces changements bénéfiques des DAPP ont fait intervenir les facteurs de transcription antioxydants (Nrf-2, PGC1α). Vraisemblablement, cette première étude a permis de démontrer la capacité des DAPP à amoindrir le SOx et à réduire l’inflammation, deux processus étroitement impliqués dans les MII. Dans la deuxième étape de mon doctorat, nous avons voulu comparer les résultats de DAPP à ceux des polyphénols dérivant de la canneberge qui est considérée par la communauté scientifique comme le fruit ayant le plus fort potentiel antioxydant. À cette fin, nous avons caractérisé l’effet des composés polyphénoliques de la canneberge (CPC) sur le SOx, la défense antioxydante et l’inflammation au niveau intestinal tout en définissant leur métabolisme intraluminal. Les différents CPC ont été séparés selon leur poids moléculaire par chromatographie et leurs structures chimiques ont été identifiées par LC-MS. Suite à une pré-incubation des cellules Caco-2/15 avec les extraits CPC (250 µg/mL), le Fe/Asc et la LPS ont été administrés comme inducteurs du SOx et de l’inflammation, respectivement. La caractérisation globale des CPC a révélé que les acides phénoliques composaient majoritairement l’extrait de canneberge de petit poids moléculaire (LC) alors que les flavonoïdes et les procyanidines dimériques/trimériques représentaient l’extrait de poids moléculaire moyen (MC) tout en laissant les procyanidines oligo et polymériques à l’extrait de haut poids moléculaire (HC). Les CPC ont permis de restaurer la plupart des perturbations engendrées dans les Caco-2/15 par le Fe/Asc et le LPS. Les CPC exhibaient le potentiel d’abaisser les niveaux de MDA, de corriger la composition des acides gras polyinsaturés n-3 et n-6, d’augmenter l’activité des enzymes antioxydantes (SOD, GPx et CAT) et d’élever l’expression de Nrf2 et PGC1α. En outre, les CPC pouvaient aussi réduire les niveaux élevés des protéines inflammatoires COX-2, TNF-α et IL-6 ainsi que la production des PGE2 par un mécanisme impliquant le NF-κB. Au niveau mitochondrial, les procyanidines oligomériques ont réussi à corriger les dysfonctions reliées à la production d’énergie (ATP), l’apoptose (Bcl-2, Cyt C et AIF) et le statut des facteurs de transcription mitochondriaux (mtTFA, mtTFB1, mtTFB2). Dans le but de bien comprendre les mécanismes d’action des CPC, nous avons défini par LC-MS les composés polyphénoliques qui ont été transportés ou absorbés par l’entérocyte. Nos analyses soulignent le transport (i) des acides cinnamiques et benzoïques (LC); (ii) la quercétine glycosylée et conjuguée et les procyanidines dimériques de type A (MC); et (iii) l’épicatéchine et les procyanidines oligomériques (HC). Les processus de métabolisation (méthylation, glucuronidation et sulfatation) au niveau de l’entérocyte ont probablement permis le transport de ces CPC surtout sous leur forme conjuguée. Les procyanidines oligomériques ayant un degré de polymérisation supérieur à 2 (HC) ont semblé adhérer aux cellules Caco-2/15. L’épicatéchine suivi par les procyanidines dimériques de type A ont été trouvés majoritaires au niveau des mitochondries. Même si nous ignorons encore l’action biologique de chaque composé polyphénolique, nous pouvons suggérer que leurs effets combinatoires exercent des fonctions antioxydantes, anti-inflammatoires et mitochondriales dans le modèle intestinal Caco-2/15. Dans une troisième étape, nous avons procédé à l’évaluation des aspects préventifs et thérapeutique des DAPP tout en sondant les mécanismes sous-jacents dans une étude préclinique. À cette fin, nous avons exploité le modèle de souris avec colite expérimentale provoquée par le Dextran Sulfate de Sodium (DSS). L’induction de l’inflammation intestinale chez la souris C57BL6 a été effectuée par l’administration orale de DSS à 2.5% pendant 10 jours. Des doses physiologiques et supra-physiologiques de DAPP (200 et 400 mg/kg/j, respectivement) ont été administrées par gavage pendant 10 jours pré- et post-DSS. L’inflammation par le DSS a provoqué une perte de poids, un raccourcissement du côlon, le décollement dystrophique de l’épithélium, l’exulcération et les infiltrations de cellules mono et polynucléaires au niveau du côlon. De plus, le DSS a induit une augmentation de la peroxidation lipidique, une régulation à la baisse des enzymes antioxydantes, une expression protéique à la hausse de la myéloperoxidase (MPO), du COX-2 et de la production des PGE2. Par ailleurs, les DAPP ont permis de corriger ou du moins d’alléger la plupart de ces anomalies en situation préventive ou thérapeutique, en plus d’abaisser l’expression protéique de NF-κB et des cytokines inflammatoires (TNF-a et l’IL-6) tout en stimulant les facteurs de transcription antioxydants (Nrf-2, PGC1α). Conséquemment, les polyphénols des DAPP ont exhibé leur puissant pouvoir antioxydant et anti-inflammatoire au niveau intestinal dans un modèle in vivo. Leurs actions sont associées à la régulation des voies de signalisation cellulaire et des changements dans la composition du microbiote. Ces trois projets de recherche permettent d’envisager l’évaluation des effets préventifs et thérapeutiques des DAPP cliniquement chez les patients avec des désordres inflammatoires de l’intestin.

Development of procedures for the triazole fungicides determination in fruits and liquid samples using microextraction techniques and chromatographic separation

Agricultural crops can be damaged by funguses, insects, worms and other organisms that cause diseases and decrease the yield of production. The effect of these damaging agents can be reduced using pesticides. Among them, triazole compounds are effective substances against fungus; for example, Oidium. Nevertheless, it has been detected that the residues of these fungicides in foods as well as in derivate products can affect the health of the consumers. Therefore, the European Union has established several regulations fixing the maximum residue of pesticide levels in a wide range of foods trying to assure the consumer safety. Hence, it is very important to develop adequate methods to determine these pesticide compounds. In most cases, gas or liquid chromatographic (GC, LC) separations are used in the analysis of the samples. But firstly, it is necessary to use proper sample treatments in order to preconcentrate and isolate the target analytes. To reach this aim, microextraction techniques are very effective tools; because allow to do both preconcentration and extraction of the analytes in one simple step that considerably reduces the source of errors. With these objectives, two remarkable techniques have been widely used during the last years: solid phase microextraction (SPME) and liquid phase microextraction (LPME) with its different options. Both techniques that avoid the use or reduce the amount of toxic solvents are convenient coupled to chromatographic equipments providing good quantitative results in a wide number of matrices and compounds. In this work simple and reliable methods have been developed using SPME and ultrasound assisted emulsification microextraction (USAEME) coupled to GC or LC for triazole fungicides determination. The proposed methods allow confidently determine triazole concentrations of μg L‐1 order in different fruit samples. Chemometric tools have been used to accomplish successful determinations. Firstly, in the selection and optimization of the variables involved in the microextraction processes; and secondly, to overcome the problems related to the overlapping peaks. Different fractional factorial designs have been used for the screening of the experimental variables; and central composite designs have been carried out to get the best experimental conditions. Trying to solve the overlapping peak problems multivariate calibration methods have been used. Parallel Factor Analysis 2 (PARAFAC2), Multivariate Curve Resolution (MCR) and Parallel Factor Analysis with Linear Dependencies (PARALIND) have been proposed, the adequate algorithms have been used according to data characteristics, and the results have been compared. Because its occurrence in Basque Country and its relevance in the production of cider and txakoli regional wines the grape and apple samples were selected. These crops are often treated with triazole compounds trying to solve the problems caused by the funguses. The peel and pulp from grape and apple, their juices and some commercial products such as musts, juice and cider have been analysed showing the adequacy of the developed methods for the triazole determination in this kind of fruit samples.

Non-fermented and fermented jabuticaba (Myrciaria cauliflora Mart.) pomaces as valuable sources of functional ingredients

abuticaba (Myrciaria cauliflora. Mart) is a highly perishable fruit native to Brazil, which is consumed both fresh and industrially processed in the form of juices, jams, wines and distilled liqueurs. This processing generates a large amount of waste by-products, which represent approximately 50% of the fruit weight. The by-products are of interest for obtaining valuable bioactive compounds that could be used as nutraceuticals or functional ingredients. In this study, fermented and non-fermented jabuticaba pomaces were studied regarding their hydrophilic and lipophilic compounds, as well as their antioxidant properties, including: soluble sugars, organic acids and tocopherols (using high performance liquid chromatography coupled to refraction index, diode array and fluorescence detector, respectively); phenolics and anthocyanins, (using liquid chromatography coupled to diode array detection, and mass spectrometry with electrospray ionization); and fatty acids (using gas-liquid chromatography with flame ionization detection). The analytical data demonstrated that jabuticaba pomaces are a rich source of bioactive compounds such as tocopherols, polyunsaturated fatty acids and phenolic compounds (namely hydrolyzable tannins and anthocyanins) with antioxidant potential. Therefore, jabuticaba pomace may have good potential as a functional ingredient in the fabrication of human foods and animal feed.

When Love Becomes Hate(?): The Interplay Between Consumer-Brand Relationships and Crisis Situations

This study had three purposes. First, it aimed to re-conceptualize organization-public relationships (OPRs) in public relations and crisis communication. This OPR re-conceptualization helps find out when the OPR buffering effect or the OPR love-becomes-hate effect happens. Second, it aimed to examine how consumer emotions are influenced by OPRs and influence consumer behavioral intentions. Third, it aimed to address the current problematic operationalization of the concept of consumer. Three pilot studies and one main study were conducted. Apple and Whole Foods were the two brands examined. One crisis that undermined the self-defining attributes shared between the brand and its consumers and another crisis that did not were examined for each brand. Almost 500 Apple consumers and 400 Whole Foods consumers provided usable questionnaires. This study had several major findings. First, non-identifying relationship and identifying relationship were different constructs. Moreover, trust, satisfaction, and commitment were not conceptually separate dimensions of OPRs. Second, the non-identifying relationships offered buffering effects by increasing positive attitudes and tempering anger and disappointment. The identifying relationships primarily offered the love-becomes-hate effects by increasing anger and disappointment. Third, if the crisis was relevant to consumers’ daily lives, brand response strategies were less effective at mitigating consumer negative reactions. Moreover, apology-compensation-reminder strategy was more effective compared to no-comment strategy. However, the apology-compensation-reminder strategy was no more effective than other strategies as long as brands compensate to the victims. Identifying relationships increased the effectiveness of response strategies. If the crisis did not undermine the self-defining attributes shared between consumers and brands, the response strategies worked even better. This study contributes to crisis communication research in multiple ways. First, it advances the OPR conceptualization by demonstrating that non-identifying relationship and identifying relationship are different concepts. More importantly, it advances the theory building of OPRs’ influences on crises by finding out when the buffering effect and the love-becomes-hate effect happen. Second, it adds to emotion research by demonstrating that strong OPRs can lead to negative emotions and positive emotions can have negative behavioral consequences on organizations. Third, the precise operationalization of the concept of consumer gives more insights about consumer reactions to crises.

Pesticide bioconcentration factor estimate in apples.

This work objective was to estimate the bioconcentration factor (BCF) of thirty six pesticides used in the Brazilian integrated apple production systems (IAP), in order to select priority pesticides to be monitored in apples. A hypothetical apple orchard was assumed and the model applied was according to Paraíba (2007) [Pesticide bioconcentration modeling for fruit trees. Chemosphere (66:1468-1475)]. The model relates BCF with plant and pesticide characteristics. The octanol-water partition coefficients of pesticides and their degradation rates in the soil were used. The following plant variables were considered: growth rate, total dry biomass, daily water transpiration rate, and total volume of water necessary to produce one kg of fresh fruit per plant. The pesticide stem-water partition coefficient and the transpiration stream concentration factor were calculated using equations that relate each coefficient with the octanol-water partition coefficient. The pesticide BCF in fruits is an important indicator of the pesticide affinity to fruits, and helps to improve the integrated production systems.

On the quality of acoustical measures when evaluating fruits quality

International audience

The impact of gender differences on consumer-brand relationships

Dissertação de Mestrado, Marketing, Faculdade de Economia, Universidade do Algarve, 2016

Integrated pest management and the (un)sustainable use of pesticides

Doutoramento em Engenharia Agronómica - Instituto Superior de Agronomia - UL

Valorização integrada de resíduos e subprodutos da extração do azeite: extração e caraterização de compostos bioativos do bagaço de azeitona

Dissertação de Mestrado, Tecnologia de Alimentos, Instituto Superior de Engenharia, Universidade do Algarve, 2016

eMed.pt - Poupe na Receita

A consulta dos preços dos medicamentos e a comparação com outros fármacos disponíveis, com preços mais baixos, é a principal funcionalidade da aplicação eMed.pt - Poupe na Receita.

Reliability of graphite furnace atomic absorption spectrometry as alternative method for trace analysis of arsenic in natural medicinal products

Purpose: To evaluate the comparative efficiency of graphite furnace atomic absorption spectrometry (GFAAS) and hydride generation atomic absorption spectrometry (HGAAS) for trace analysis of arsenic (As) in natural herbal products (NHPs). Method: Arsenic analysis in natural herbal products and standard reference material was conducted using atomic absorption spectrometry (AAS), namely, hydride generation AAS (HGAAS) and graphite furnace (GFAAS). The samples were digested with HNO3–H2O2 in a ratio of 4:1 using microwaveassisted acid digestion. The methods were validated with the aid of the standard reference material 1515 Apple Leaves (SRM) from NIST Results: Mean recovery of three different samples of NHPs, using HGAAS and GFAAS, ranged from 89.3 - 91.4 %, and 91.7 - 93.0 %, respectively. The difference between the two methods was insignificant. A (P= 0.5), B (P=0.4) and C (P=0.88) Relative standard deviation (RSD) RSD, i.e., precision was 2.5 - 6.5 % and 2.3 - 6.7 % using HGAAS and GFAAS techniques, respectively. Recovery of arsenic in SRM was 98 and 102 % by GFAAS and HGAAS, respectively. Conclusion: GFAAS demonstrates acceptable levels of precision and accuracy. Both techniques possess comparable accuracy and repeatability. Thus, the two methods are recommended as an alternative approach for trace analysis of arsenic in natural herbal products.

Natural parasitism of lepidopteran eggs by Trichogramma species (Hymenoptera: Trichogrammatidae) in agricultural crops in Minas Gerais, Brazil.

The genus Trichogramma Westwood (Hymenoptera: Trichogrammatidae) includes insect egg parasitoids that are widely used throughout the world as control agents of pest insects. The aim of this study was to identify the species of Trichogramma naturally associated with the eggs of lepidopteran pests of the following agricultural and horticultural crops: collards, Brassica oleracea L. (Brassicales: Brassicaceae); papaya, Carica papaya L. (Capparales: Caricaceae); tomato, Lycopersicon esculentum Mill. (Solanales: Solanaceae); cassava, Manihot esculenta Crantz (Malpighiales: Euphorbiaceae); banana, Musa sp. L. (Zingiberales: Musaceae); passion fruit, Passiflora sp. Degener (Malpighiales: Passifloraceae); sugarcane, Saccharum sp. L. (Poales: Poaceae); and corn (maize), Zea mays L. (Poales: Poaceae); and an invasive species (Sodom?s apple milkweed, Calotropis procera Aiton; Gentianales: Apocynaceae) in the semiarid region of Minas Gerais, Brazil. We report natural parasitism by Trichogramma in eggs of Agraulis vanillae vanillae (L.) (Lepidoptera: Nymphalidae), Antichloris eriphia F. (Lepidoptera: Arctiidae), Danaus sp. (L.) (Lepidoptera: Nymphalidae), Diatraea saccharalis F. (Lepidoptera: Crambidae), Erinnyis ello L. (Lepidopera: Sphingidae), and Protambulyx strigilis L. (Lepidopera: Sphingidae). In total, 2,242 specimens of Trichogramma were obtained, belonging to the species T. pretiosum Riley, T. manicobai Brun, Moraes & Soares, T. marandobai Brun, Moraes & Soares, and T. galloi Zucchi. These species of Trichogramma may be candidates for biological control programs of lepidopteran pests in the semiarid region of Minas Gerais and in other semiarid regions.

The empty child: the reproduction of culture through schooling

This research provides a needed glimpse at the ‘underside’ (Giroux, 1981) of Australian pedagogy.By charting the lives of six teachers and six students from a range of Victorian schools, the research examines our assumptions about how identities are produced and reproduced through schooling. It provides a detailed analysis through a combination of photo elicitation and narrative inquiry methods. The study examines the resistance to sameness and why some students and teachers submit and accept the sameness, while others push back.Building initially from the work of Michael Apple and his positioning of ideology, curriculum and power relations, this study explores what are the values and assumptions that shape pedagogical practice and in what ways do these practices inculcate Victorian students into ideologically ridden‘common sense’ practices that are not articulated but understood through the consumption of cultural capital?This research seeks access to the powerful undercurrents that shape the identities of our teachers and our students as ‘foot soldiers in the long front of modernity’ (Willis, 2003, p. 390). The research is concerned with schooling and the process that happens to an individual within a school. It examines power and education and asks how do both students and teachers internalize and express the pedagogies of contemporary schooling and in what ways do the everyday aspects of teaching and learning reinforce the notion of schools as mechanisms of cultural capital distribution?

Do physical activity and dietary smartphone applications incorporate evidence-based behaviour change techniques?

BACKGROUND: There has been a recent proliferation in the development of smartphone applications (apps) aimed at modifying various health behaviours. While interventions that incorporate behaviour change techniques (BCTs) have been associated with greater effectiveness, it is not clear to what extent smartphone apps incorporate such techniques. The purpose of this study was to investigate the presence of BCTs in physical activity and dietary apps and determine how reliably the taxonomy checklist can be used to identify BCTs in smartphone apps.

METHODS: The top-20 paid and top-20 free physical activity and/or dietary behaviour apps from the New Zealand Apple App Store Health & Fitness category were downloaded to an iPhone. Four independent raters user-tested and coded each app for the presence/absence of BCTs using the taxonomy of behaviour change techniques (26 BCTs in total). The number of BCTs included in the 40 apps was calculated. Krippendorff's alpha was used to evaluate interrater reliability for each of the 26 BCTs.

RESULTS: Apps included an average of 8.1 (range 2-18) techniques, the number being slightly higher for paid (M = 9.7, range 2-18) than free apps (M = 6.6, range 3-14). The most frequently included BCTs were "provide instruction" (83% of the apps), "set graded tasks" (70%), and "prompt self-monitoring" (60%). Techniques such as "teach to use prompts/cues", "agree on behavioural contract", "relapse prevention" and "time management" were not present in the apps reviewed. Interrater reliability coefficients ranged from 0.1 to 0.9 (Mean 0.6, SD = 0.2).

CONCLUSIONS: Presence of BCTs varied by app type and price; however, BCTs associated with increased intervention effectiveness were in general more common in paid apps. The taxonomy checklist can be used by independent raters to reliably identify BCTs in physical activity and dietary behaviour smartphone apps.

Building a local design and entrepreneurship ecosystem

Design and creativity are becoming greatly sought out skills in leading industries around the world, big businesses are developing the “Chief Design Officer” to engage with strategic and company shaping discussions. Design as an economic driver is now abundantly clear with companies such as Nike and Apple leading this way of thinking, but how do we as Australian industry capture this and how do we instil “creativity” into our secondary school and university level education to drive the next level of innovation and development. The local region where Deakin University is situated has undergone significant changes in the last 10 years, what was once an economy dominated by oil, automotive and metal production industries has been wound down to a local economy dominated by health, services and education. However, manufacturing and design being the front end of manufacturing is still a key economic driver this study is looking at the embryonic initiatives undertaken to build an ecosystem of design and entrepreneurship in a regional area. Several aspects will be looked at, high school and university student engagement in the process, established SME's and start-up culture. With the establishment of an ecosystem it is believed that success will breed success. With student engagement showing that being creative and playing can yield tangible results, it also gets students comfortable with the element of risk. The efforts of Deakin University is about providing the framework and scaffolding for students to pursue a start-up idea and test it validity. The final part of the ecosystem is for SME's and recent start-ups to share their success stories and acting as mentors as future start-ups emerge. By creating an ecosystem that is driven by design, manufacturing and entrepreneurship key economic outcomes will be generated; a regional area will be more resilient to economic uncertainty and ultimately a cohort of innovative thinkers that will generate value for their community.