Different routes to pure food allergens: extraction, recombinant techniques and total chemical synthesis for obtaining the plant-food pan-allergen LTP

La richiesta di allergeni puri è in continuo aumento per scopi diagnostici, come standard per metodi di rilevamento e di quantificazione, per l'immunoterapia e per lo studio a livello molecolare dei meccanismi delle reazioni allergiche, al fine di facilitare lo sviluppo di possibili cure. In questa tesi di dottorato sono descritte diverse strategie per l’ottenimento di forme pure di non-specific Lipid Transfer Proteins (nsLTPs), le quali sono state riconosciute essere rilevanti allergeni alimentari in molti frutti e verdure comunemente consumati e sono state definite come modello di veri allergeni alimentari. Una LTP potenzialmente allergenica, non nota in precedenza, è stata isolata dalle mandorle, mentre una LTP dall’allergenicità nota contenuta nelle noci è stata prodotta mediante tecniche di DNA ricombinante. Oltre a questi approcci classici, metodi per la sintesi chimica totale di proteine sono stati applicati per la prima volta alla produzione di un allergene, utilizzando Pru p 3, la LTP prototipica e principale allergene della pesca nell'area mediterranea, come modello. La sintesi chimica totale di proteinepermette di controllarne completamente la sequenza e di studiare la loro funzione a livello atomico. La sua applicazione alla produzione di allergeni costituisce perciò un importante passo avanti nel campo della ricerca sulle allergie alimentari. La proteina Pru p 3 è stata prodotta nella sua intera lunghezza e sono necessari solo due passaggi finali di deprotezione per ottenere il target nella sua forma nativa. Le condizioni sperimentali per tali deprotezioni sono state messe a punto durante la produzione dei peptidi sPru p 3 (1-37) e sPru p 3 (38-91), componenti insieme l'intera proteina. Tecniche avanzate di spettrometria di massa sono state usate per caratterizzare tutti i composti ottenuti, mentre la loro allergenicità è stata studiata attraverso test immunologici o approcci in silico.

Lifestyle Behaviors and Self-Rated Health: The Living for Health Program

Background. Lack of adherence to dietary and physical activity guidelines has been linked to an increase in chronic diseases in the United States (US). The aim of this study was to assess the association of lifestyle behaviors with self-rated health (SRH). Methods. This cross-sectional study used self-reported data from Living for Health Program ( 1,701) which was conducted from 2008 to 2012 in 190 health fair events in South Florida, US. Results. Significantly higher percent of females as compared to males were classified as obese (35.4% versus 27.0%), reported poor/fair SRH (23.4% versus 15.0%), and were less physically active (33.9% versus 25.4%). Adjusted logistic regression models indicated that both females and males were more likely to report poor/fair SRH if they consumed 2 servings of fruits and vegetables per day (, 95% CI 1.30–3.54; , 95% CI 1.12–7.35, resp.) and consumed mostly high fat foods (, 95% CI 1.03–2.43; , 95% CI 1.67–2.43, resp.). The association of SRH with less physical activity was only significant in females (, 95% CI 1.17–2.35). Conclusion. Gender differences in health behaviors should be considered in designing and monitoring lifestyle interventions to prevent cardiovascular diseases.

Design and development of a diagonal-airflow batch dryer for spatial drying homogeneity

Das Verfahren der Lebensmitteltrocknung wird häufig angewendet, um ein Produkt für längere Zeit haltbar zu machen. Obst und Gemüse sind aufgrund ihres hohen Wassergehalts leicht verderblich durch biochemische Vorgänge innerhalb des Produktes, nicht sachgemäße Lagerung und unzureichende Transportmöglichkeiten. Um solche Verluste zu vermeiden wird die direkte Trocknung eingesetzt, welche die älteste Methode zum langfristigen haltbarmachen ist. Diese Methode ist jedoch veraltet und kann den heutigen Herausforderungen nicht gerecht werden. In der vorliegenden Arbeit wurde ein neuer Chargentrockner, mit diagonalem Luftstömungskanal entlang der Länge des Trocknungsraumes und ohne Leitbleche entwickelt. Neben dem unbestreitbaren Nutzen der Verwendung von Leitblechen, erhöhen diese jedoch die Konstruktionskosten und führen auch zu einer Erhöhung des Druckverlustes. Dadurch wird im Trocknungsprozess mehr Energie verbraucht. Um eine räumlich gleichmäßige Trocknung ohne Leitbleche zu erreichen, wurden die Lebensmittelbehälter diagonal entlang der Länge des Trockners platziert. Das vorrangige Ziel des diagonalen Kanals war, die einströmende, warme Luft gleichmäßig auf das gesamte Produkt auszurichten. Die Simulation des Luftstroms wurde mit ANSYS-Fluent in der ANSYS Workbench Plattform durchgeführt. Zwei verschiedene Geometrien der Trocknungskammer, diagonal und nicht diagonal, wurden modelliert und die Ergebnisse für eine gleichmäßige Luftverteilung aus dem diagonalen Luftströmungsdesign erhalten. Es wurde eine Reihe von Experimenten durchgeführt, um das Design zu bewerten. Kartoffelscheiben dienten als Trocknungsgut. Die statistischen Ergebnisse zeigen einen guten Korrelationskoeffizienten für die Luftstromverteilung (87,09%) zwischen dem durchschnittlich vorhergesagten und der durchschnittlichen gemessenen Strömungsgeschwindigkeit. Um den Effekt der gleichmäßigen Luftverteilung auf die Veränderung der Qualität zu bewerten, wurde die Farbe des Produktes, entlang der gesamten Länge der Trocknungskammer kontaktfrei im on-line-Verfahren bestimmt. Zu diesem Zweck wurde eine Imaging-Box, bestehend aus Kamera und Beleuchtung entwickelt. Räumliche Unterschiede dieses Qualitätsparameters wurden als Kriterium gewählt, um die gleichmäßige Trocknungsqualität in der Trocknungskammer zu bewerten. Entscheidend beim Lebensmittel-Chargentrockner ist sein Energieverbrauch. Dafür wurden thermodynamische Analysen des Trockners durchgeführt. Die Energieeffizienz des Systems wurde unter den gewählten Trocknungsbedingungen mit 50,16% kalkuliert. Die durchschnittlich genutzten Energie in Form von Elektrizität zur Herstellung von 1kg getrockneter Kartoffeln wurde mit weniger als 16,24 MJ/kg und weniger als 4,78 MJ/kg Wasser zum verdampfen bei einer sehr hohen Temperatur von jeweils 65°C und Scheibendicken von 5mm kalkuliert. Die Energie- und Exergieanalysen für diagonale Chargentrockner wurden zudem mit denen anderer Chargentrockner verglichen. Die Auswahl von Trocknungstemperatur, Massenflussrate der Trocknungsluft, Trocknerkapazität und Heiztyp sind die wichtigen Parameter zur Bewertung der genutzten Energie von Chargentrocknern. Die Entwicklung des diagonalen Chargentrockners ist eine nützliche und effektive Möglichkeit um dei Trocknungshomogenität zu erhöhen. Das Design erlaubt es, das gesamte Produkt in der Trocknungskammer gleichmäßigen Luftverhältnissen auszusetzen, statt die Luft von einer Horde zur nächsten zu leiten.

Analysis of the Market Potential of Syrian Organic Fruit and Vegetables for Exports to Germany

Syria has been a major producer and exporter of fresh fruit and vegetables (FFV) in the Arabic region. Prior to 2011, Syrian FFV were mainly exported to the neighbouring countries, the Gulf States and Northern Africa as well as to Eastern European countries. Although the EU is potentially one of the most profitable markets of high quality FFV (such as organic ones) in the world, Syrian exports of FFV to Western European countries like Germany have been small. It could be a lucrative opportunity for Syrian growers and exporters of FFV to export organic products to markets such as Germany, where national production is limited to a few months due to climatic conditions. Yet, the organic sector in Syria is comparatively young and only a very small area of FFV is certified according to EU organic regulations. Up to the author’s knowledge, little was known about Syrian farmers’ attitudes towards organic FFV production. There was also no study so far that explored and analysed the determining factors for organic FFV adoption among Syrian farmers as well as the exports of these products to the EU markets. The overarching aim of the present dissertation focused on exploring and identifying the market potential of Syrian exports of organic FFV to Germany. The dissertation was therefore concerned with three main objectives: (i) to explore if German importers and wholesalers of organic FFV see market opportunities for Syrian organic products and what requirements in terms of quality and quantity they have, (ii) to determine the obstacles Syrian producers and exporters face when exporting agricultural products to Germany, and (iii) to investigate whether Syrian farmers of FFV can imagine converting their farms to organic production as well as the underlying reasons why they do so or not. A twofold methodological approach with expert interviews and a farmer survey were used in this dissertation to address the abovementioned objectives. While expert interviews were conducted with German and Syrian wholesalers of (organic) FFV in 2011 (9 interviews each), the farmer survey was administrated with 266 Syrian farmers of FFV in the main region for the production of FFV (i.e. the coastal region) from November 2012 till May 2013. For modelling farmers’ decisions to adopt organic farming, the Theory of Planned Behaviour as theoretical framework and Partial Least Squares Structural Equation Modelling as the main method for data analysis were used in this study. The findings of this dissertation yield implications for the different stakeholders (governmental institutions and NGOs, farmers, exporters, wholesalers, etc.) who are interested in prompting the Syrian export of organic products. Based on the empirical results and a literature review, an action plan to promote Syrian production and export of organic products was developed which can help in the post-war period in Syria at improving the organic sector.

Gardening for Health: Patterns of Gardening and Fruit and Vegetable Consumption on the Navajo Nation

Thesis (Master's)--University of Washington, 2016-08

Enhancing the total phenolic content and antioxidants of lemon pomace aqueous extracts by applying UV-C irradiation on the dried powder

Several studies have shown that UV-C irradiation promotes the bioactive compounds and antioxidants of fresh fruits and vegetables. The aim of this study was to apply UV irradiation in lemon pomace dried powder for enhancing its phenolic content and antioxidant properties, thus more bioactive compounds should be available for extraction and utilization. Lemon pomace dried powder was placed above the UV lamp and treated with dosages of 4, 19, 80 and 185 kJ m-2, while untreated powder was used as a control. UV-C irradiation significantly affected the total phenolic content, total flavonoid content, proanthocyanidins and antioxidant capacity measured by CUPRAC and FRAP of the lemon pomace dried powder, while it did not affect the vitamin C content. UV-C irradiation of 19 kJ m-2 resulted in 19% higher total phenolic content than the control, while UV-C irradiation of 180 kJ m-2 resulted in 28% higher total flavonoid content than the control. The antioxidant capacity was reduced when UV-C irradiation more than 4 kJ m-2 was applied. The results of this study indicate that UV-C treatment has the potential to increase the extraction of bioactive compounds of lemon dried pomace at relatively high dosages.

Effect of proteolytic enzyme and fiber of papaya fruit on human digestive health

The World Health Organization (WHO, 2005) recommends consumption of fruits and vegetables as part of a healthy diet with daily recommendation of 5 servings or at least 400 g per day. Fruits and vegetables are good sources of vitamins, minerals, antioxidants, and fiber. Papaya fruit is known for his high nutrient and fiber content, and with few exceptions, it is generally consumed ripe due to its characteristic flavor and aroma. Digestion improvement has been attributed to consumption of papaya; this we speculate is attributed to the fiber content and proteolytic enzymes associated with this highly nutritious fruit. However, research is lacking that evaluates the impact of papaya fruit on human digestion. Papain is a proteolytic enzyme generally extracted from the latex of unripe papaya. Previous research has focused on evaluating papain activity from the latex of different parts of the plant; however there are no reports about papain activity in papaya pulp through fruit maturation. The activity of papain through different stages of ripeness of papaya and its capacity of dislodging meat bolus in an in vitro model was addressed. The objective of this study was to investigate whether papain activity and fiber content are responsible for the digestive properties attributed to papaya and to find a processing method that preserves papaya health properties with minimal impact on flavor. Our results indicated that papain was active at all maturation stages of the fruit. Ripe papaya pulp displayed the highest enzyme activity and also presented the largest meat bolus displacement. The in vitro digestion study indicated that ripe papaya displayed the highest protein digestibility; this is associated with proteolytic enzymes still active at the acidity of the stomach. Results from the in vitro fermentation study indicated that ripe papaya produced the highest amount of Short Chain Fatty Acids SCFA of the three papaya substrates (unripe, ripe, and processed). SCFA are the most important product of fermentation and are used as indicators of the amount of substrate fermented by microorganisms in the colon. The combination of proteolytic enzymes and fiber content found in papaya make of this fruit not only a potential digestive aid, but also a good source of SCFA and their associated potential health benefits. Irradiation processing had minimal impact on flavor compounds of papaya nectar. However, processed papaya experienced the lowest protein digestibility and SCFA production among the papaya substrates. Future research needs to explore new processing methods for papaya that minimize the detrimental impact on enzyme activity and SCFA production.

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.

Myths and legends of flowers, trees, fruits, and plants in all ages and in all climes,

"Second edition."

Inequities in energy-balance related behaviours and family environmental determinants in European children: baseline results of the prospective EPHE evaluation study

BACKGROUND:Tackling inequalities in overweight, obesity and related determinants has become a top priority for the European research and policy agendas. Although it has been established that such inequalities accumulate from early childhood onward, they have not been studied extensively in children. The current article discusses the results of an explorative analysis for the identification of inequalities in behaviours and their determinants between groups with high and low socio-economic status. METHODS: This study is part of the Epode for the Promotion of Health Equity (EPHE) evaluation study, the overall aim of which is to assess the impact and sustainability of EPODE methodology to diminish inequalities in childhood obesity and overweight. Seven community-based programmes from different European countries (Belgium, Bulgaria, France, Greece, Portugal, Romania, The Netherlands) participate in the EPHE study. In each of the communities, children aged 6-8 years participated, resulting in a total sample of 1266 children and their families. A parental self-administrated questionnaire was disseminated in order to assess the socio-economic status of the household, selected energy balance-related behaviours (1. fruit and vegetable consumption; 2. soft drink/ fruit juices and water consumption; 3. screen time and 4. sleep duration) of the children and associated family environmental determinants. The Mann-Whitney U test and Pearson's chi-square test were used to test differences between the low and high education groups. The country-specific median was chosen as the cut-off point to determine the educational level, given the different average educational level in every country. RESULTS: Children with mothers of relatively high educational level consumed fruits and vegetables more frequently than their peers of low socio-economic status. The latter group of children had a higher intake of fruit juices and/or soft drinks and had higher screen time. Parental rules and home availability were consistently different between the two socio-economic groups in our study in all countries. However we did not find a common pattern for all behaviours and the variability across the countries was large. CONCLUSIONS: Our findings are indicative of socio-economic inequalities in our samples, although the variability across the countries was large. The effectiveness of interventions aimed at chancing parental rules and behaviour on health inequalities should be studied.

Neotropical Copestylum Macquart (Diptera: Syrphidae) Breeding in Fruits and Flowers, Including 7 New Species

Ten species of Copestylum (Diptera: Syrphidae) were reared from fruits and flowers in Costa Rica, Ecuador and Trinidad. Seven were new and in this paper, we describe them, their development sites and the third stage larva and/or the puparium of all ten species. One new synonym is proposed, Copestylum pinkusi (Curran) [= Copestylum cinctiventre (Curran)]. Similarities and differences between these new and other Copestylum species, suggest they separate into two groups, referred to as the Vagum and Cinctiventre species groups. Features characterising these groups for both adult and early stages are assessed. Each species was also distinguished using adult and early stage characters. Within the Vagum group, adults were more disparate morphologically than the larval stage; this was reversed in the Cinctiventre group. Adult colour patterns are probably cryptic in function and for disguise. Vagum species have disruptive marks, while the Cinctiventre species have reflective colours. Biologically, the groups are almost distinguished by larval development sites. Vagum species use predominantly fruits and have a larval stage that is relatively generalised in form and habit. Cinctiventre species are confined to developing in flowers and the larva is more specialised. A key to both adult and early stages of all ten species is provided.

Bioactive Phenols in Small Fruits and Berries

Bioactive compounds are extra nutritional constituents occurring naturally in plant foods in small amounts, however in quantities enough to produce bioactive effects. Among bioactive compounds the phenolic compounds are a very large set of molecules, which include several groups such as for example flavonoids, phenolic acids or tannins. Small fruits and berries include a wide diversity of fruits, like grapes, strawberries, blackberries, blueberries, raspberries, cherries, hardi kiwi, gooseberries, cranberries, currants (black, white, red), physalis, crowberries, açaí, elderberries, dates or goji berries, and these frequently have been reported as having particularly high concentrations of phenolic compounds with antioxidant activity. Hence, the objective of this chapter is to review the literature about the type and contents of different phenolic compounds present in small fruits and berries, as well as their bioactive properties, including antioxidant capacity. All the fruits and berries investigated in this chapter were particularly rich in bioactive compounds, including phenolic compounds that provide the fruits with high antioxidant properties. The most relevant health promoting effects include anti-cancer, anti-inflamatory, neuro protective, cardio protective or anti-diabetes, thus indicating that these foods are a valuable resource to prevent and treat diseases.

Oral food perception and polyphenol-rich foods acceptance - the importance of knowing individuals saliva characteristics for promoting consumption

Polyphenols are widely present in fruits, vegetables, cereals and beverages. Their study gained scientific interest because of their beneficial effects on health. Although there is currently no official dietary recommendation for polyphenol intake, health professionals recommend the consumption of 5-8 daily portions of fruits and vegetables. This is not always achieved and, despite possible causes associated to practical schedule difficulties, the aversive bitter and astringent sensations associated to polyphenols may also lead to avoidance. As such, a better understanding on mechanisms responsible for differences among people, in polyphenol oral perception, is needed for promoting healthier choices. Saliva has been linked to polyphenol consumption. We have previously observed, in animal models, changes in salivary proteome induced by tannin-enriched diets. Moreover, differences in astringency perception were attributed to differences in salivary protein composition. In a recent experiment, we observed differences among individuals with dissimilar tannic-acid perception: people with high sensitivity for the oral sensations elicited by tannins have higher amounts of salivary cystatins and lower capacity to maintain their levels after tannic-acid ingestion. Additionally, and similarly to previous studies, salivary amylase was observed to be involved in tannin perception. In this presentation, oral cavity characteristics influencing the perception of polyphenol-containing foods will be discussed.

Exercise alone is not enough : weight loss also needs a healthy (Mediterranean) diet?

Objective: In the majority of exercise intervention studies, the aggregate reported weight loss is often small. The efficacy of exercise as a weight loss tool remains in question. The aim of the present study was to investigate the variability in appetite and body weight when participants engaged in a supervised and monitored exercise programme. ---------- Design: Fifty-eight obese men and women (BMI = 31·8 ± 4·5 kg/m2) were prescribed exercise to expend approximately 2092 kJ (500 kcal) per session, five times a week at an intensity of 70 % maximum heart rate for 12 weeks under supervised conditions in the research unit. Body weight and composition, total daily energy intake and various health markers were measured at weeks 0, 4, 8 and 12. ---------- Results: Mean reduction in body weight (3·2 ± 1·98 kg) was significant (P < 0·001); however, there was large individual variability (−14·7 to +2·7 kg). This large variability could be largely attributed to the differences in energy intake over the 12-week intervention. Those participants who failed to lose meaningful weight increased their food intake and reduced intake of fruits and vegetables. ---------- Conclusion: These data have demonstrated that even when exercise energy expenditure is high, a healthy diet is still required for weight loss to occur in many people.