Proteomic analysis reveals suppression of bark chitinases and proteinase inhibitors in citrus plants affected by the citrus sudden death disease

Citrus sudden death (CSD) is a disease of unknown etiology that greatly affects sweet oranges grafted on Rangpur lime rootstock, the most important rootstock in Brazilian citriculture. We performed a proteomic analysis to generate information related to this plant pathogen interaction. Protein profiles from healthy, CSD-affected and CSD-tolerant stem barks, were generated using two-dimensional gel electrophoresis. The protein spots were well distributed over a pI range of 3.26 to 9.97 and a molecular weight (MW) range from 7.1 to 120 kDa. The patterns of expressed proteins on 2-DE gels made it possible to distinguish healthy barks from CSD-affected barks. Protein spots with MW around 30 kDa and pI values ranging from 4.5 to 5.2 were down-regulated in the CSD-affected rootstock bark. This set of protein spots was identified as chitinases. Another set of proteins, ranging in pI from 6.1 to 9.6 with an MW of about 20 kDa, were also suppressed in CSD-affected rootstock bark; these were identified as miraculin-like proteins, potential trypsin inhibitors. Downregulation of chitinases and proteinase inhibitors in CSD-affected plants is relevant since chitinases are well-known pathogenesis-related protein, and their activity against plant pathogens is largely accepted.

Nuclear magnetic resonance characterization of metabolite disorder in orange trees caused by citrus sudden death disease

Citrus sudden death (CSD) is a new disease of sweet orange and mandarin trees grafted on Rangpur lime and Citrus volkameriana rootstocks. It was first seen in Brazil in 1999, and has since been detected in more than four million trees. The CSD causal agent is unknown and the current hypothesis involves a virus similar to Citrus tristeza virus or a new virus named Citrus sudden death-associated virus. CSD symptoms include generalized foliar discoloration, defoliation and root death, and, in most cases, it can cause tree death. One of the unique characteristics of CSD disease is the presence of a yellow stain in the rootstock bark near the bud union. This region also undergoes profound anatomical changes. In this study, we analyse the metabolic disorder caused by CSD in the bark of sweet orange grafted on Rangpur lime by nuclear magnetic resonance (NMR) spectroscopy and imaging. The imaging results show the presence of a large amount of non-functional phloem in the rootstock bark of affected plants. The spectroscopic analysis shows a high content of triacylglyceride and sucrose, which may be related to phloem blockage close to the bud union. We also propose that, without knowing the causal CSD agent, the determination of oil content in rootstock bark by low-resolution NMR can be used as a complementary method for CSD diagnosis, screening about 300 samples per hour.

Konstruktion och design av två glassförpackningar i kartongkvaliteten Frövi Light

This degree project was made in cooperation with AssiDomän Frövi and Charlotte Andersson and the main objectivehas been to create to ice cream pacbges from 250 gsm Frövi Light board. The paekages are intended to be easilyresealable while decreasing in size together with the ice cream. The project also describes the food packaging labelsand symbols currently present in the European common market.Both capsules utilize a separate lid for resealing. One of the paekages (Capsuie A) is indended to be cut along withthe ice cream and thereby decrease in size. The other one (Capsuie B) uses a series of flaps for contracting and expandingwhich enab1es adjusting of the size without cutting the package up.The design for Capsuie A is created both as a series of flavours with a super-hero theme intended for children andwith a colfee flavour for a maturer audience. For Capsule B, a design was created for a sorbet.

Hur retusch påverkar skönhetsannonsers attraktion : En studie om målgruppens åsikter och köplust

Tidigare studier som gjorts inom området visade att kvinnor anser att kvinnors skönhet inte är värderad enligt rimliga normer i vårt samhälle. Hård retuschering motarbetas på många håll, bland annat genom att organisationer som ASA (Advertising Standards Authority) och politiska partier som Liberaldemokraterna i Storbritannien arbetar för att missledande retuschering ska försvinna. En studie har visat att det, för att öka annonsers attraktionskraft, är viktigt att den ökade skönheten hos en modell är relevant för produktens användningsområde, då betraktaren anstränger sig intellektuellt vid betraktandet av bilden. Det finns dock inga tidigare studier som bevisar om en retuscherad modell faktiskt ökar målgruppens köplust av skönhetsprodukter vid rådande samhällsattityder och skönhetsideal. Två enkätundersökningar gjordes därför med sammanlagt 1480 deltagare. Produkten i studien var en fiktiv hudkräm. Genom att i den ena undersökningen låta betraktaren möta modellbilden i en situation som efterliknade verkligheten, på så sätt att de inte fick veta om den var retuscherad, kunde studien svara på om retuschering ökade köplusten. Bilder med bortretuscherade skönhetsfläckar och utväxt prefererades. Att retuschera bort flyghår ökade inte köplusten. I den andra jämförande undersökningen kunde betraktaren jämföra den retuscherade med den oretuscherade modellbilden. Resultaten visade att skönhetsfläckar och flyghår tillåts och prefereras borttagna, men inte utväxt. I intervjuer med fem kvinnor hördes målgruppens attityder om vad som ansågs var tillåtet att retuschera. Svaren tydde på att en relevant och försiktig retuschering prefererades. Att lämna några födelsemärken ökade trovärdigheten.

Ensamhet - gemenskap : Ett spänningsfält i Vilhelm Ekelunds författarskap

For the Swedish poet, essayist and aphorist Vilhelm Ekelund, ensamhet (solitude) and gemenskap (intellectual and spiritual community) were highly complex notions, with various and often contradictory meanings. In this article, I argue that both concepts have positive as well as negative connotations in Ekelund’s texts. Solitude can be sweet and delightful and the poet/writer may long for it, but it can also appear to him as a sordid and painful state. In the same way, life with other people may be just as difficult and complicated. I show that Ekelund as a young poet both embraced solitude as a positive notion and suffered from depressing isolation. The theme of solitude also appeared in his early prose as a heroic stance fitting for an extraordinary person. According to Ekelund, the fate of the truly gifted artist is loneliness, and he will find great difficulties connecting with people around him. In fact, he will find intellectual and spiritual community only when communicating with the great precursors – in Ekelund’s case that meant the prominent figures of Greek and Roman cultural heritage. “Modern” artists interested him only in so much as they openly venerated this classicist tradition. Ekelund may have despaired at the idea of an intellectual or spiritual community with his contemporaries; he was, nevertheless, optimistic regarding the ability of later generations to understand him. He was convinced that he did not write for people in his own time but, indeed, for posterity.

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Avaliação de cultivares de melancia na Depressão Central do Rio Grande do Sul

O objetivo do presente trabalho conduzido na Estação Experimental Agronômica da Universidade Federal do Rio Grande do Sul em Eldorado do Sul, RS, em um solo Podzólico Vermelho Amarelo, foi avaliar nove cultivares de melancia (Citrullus lanatus) buscando identificar cultivares de melhor adaptação à região, possibilitando o cultivo em anos sucessivos na mesma área, pois os produtores devido a problemas fitossanitários buscam a cada cultivo novas áreas para produzir. A semeadura no espaçamento 3,0m x 1,5m, foi feita em novembro, com ressemeadura em dezembro de 2000. As cultivares foram avaliadas quanto à produtividade e ocorrência de doenças. Em termos de produtividade, a cultivar Vista F1, com 10,8 t.ha-1, foi significativamente mais produtiva que a cultivar Vitória F1 (2,6 t.ha-1). Quanto a ocorrência de doenças, a cultivar Athens F1 apresentou o menor número de plantas com sintomas de antracnose (Colletotrichum orbiculare), com 5,4 plantas, diferindo estatisticamente das cultivares Vista F1, Crimson Select, Crimson Magic F1, com 9,2, 8,6, 8,5 plantas, respectivamente, e das cultivares Vitória F1, Arriba F1 e Verona F1, com 8,1 plantas com sintomas. O peso 6,4 Kg por fruto da cultivar Crimson Magic F1 foi significativamente maior que o peso de frutos das cultivares Vitória F1 (4,4 Kg) e Crimson Sweet (4,4 Kg). O peso de melancias das demais cultivares foi intermediário e não diferiu estatisticamente do peso médio da cultivar Crimson Magic F1. Na classificação dos frutos por peso, somente as cultivares Athens F1 e Vista F1 apresentaram frutos com peso acima de 10 Kg, frutos na categoria especial. As cultivares Verona F1, Crimson Magic F1 e Vista F1 apresentaram a melhor performance em relação ao número de frutos na categoria 1, isto é, frutos com peso entre 6 e 10 Kg . As cultivares Vista F1 e Crimson Magic F1 destacaram-se na produtividade e a cultivar Athens F1 destacou-se na resistência a antracnose. Estas cultivares são promissoras, mas são necessários novos ensaios para consolidar estes resultados.

Avaliação da produtividade de biomassa de capim elefante e sorgo sacarino no Estado do Ceará para uso energético

Com a crescente elevação nas possibilidades de utilização da biomassa nos mais diversos setores da indústria, o presente estudo visa identificar uma alternativa para aumentar a produção utilizando, para tanto, o capim elefante (Pennisetum purpureum) e sorgo sacarino (Sorghum bicolor L.). Os experimentos foram realizados na Fazenda Vitória (4°21'36"S 38°5'17"W) em Beberibe – CE, em uma área total de 1.944 m2. A semeadura foi realizada em 36 parcelas de 54 m2, em delineamento inteiramente casualizado, sendo 9 tratamentos (genótipos) e 4 repetições. Foram utilizados seis diferentes genótipos de sorgo sacarino, s_1, s_2, s_3, s_4, s_5 e s_6. Fazem parte do estudo, ainda, um híbrido de capim elefante e milheto (Pennisetum glaucum), comercialmente conhecido como paraíso, e dois tratamentos de capim elefante, Cameroon e Napier. Para o sorgo sacarino, foram realizadas três colheitas, a primeira avaliada semanalmente e as demais com avaliação única. Para o capim elefante foram realizadas duas colheitas, a primeira com intervalo de 186 dias e a segunda com intervalo de 92 dias. Esse intervalo possibilitou a comparação da produtividade de biomassa de cada tratamento de forma trimestral e semestral. Os resultados observados mostraram que os tratamentos de maior destaque de produtividade de biomassa (base seca) foram com capim elefante, independente do período. Com base nessas informações, concluiu-se que, analisando apenas a produtividade de biomassa, o mais indicado dos tratamentos estudados é o capim elefante Napier ou Cameroon. No entanto, constatou-se que o sorgo sacarino apresentou grande potencial de produtividade com um aditivo, seu mosto pode ser utilizado no processo de fermentação para obtenção do etanol.

Effect of the estufagem process on the chemical constituents of Madeira wines

Madeira wine is a product of well-established reputation, whose aroma and flavour is the result of unique combinations. Particularly, its maturation may include estufagem, wherein wine is usually heated at 45 °C for three months. During this period, several chemical changes may occur, so it is essential to assess its impact on the wine. In this sense, the main objective of the thesis was to evaluate the effect estufagem on the chemical constituents of Madeira wine, specifically on those molecules potentially important in the development of its typical features. Firstly, analytical methodologies capable of determining the target compounds, combining precision and reproducibility to execution effectiveness, were developed. Then various monovarietal Madeira wines were analysed during estufagem under standard and overheating conditions in order to assess its effect. The following compounds were evaluated: furans, amino acids, biogenic amines, polyphenols, organic acids and volatile compounds. In addition, the total polyphenolic composition, the antioxidant potential and the colour of these wines were also evaluated. The results show that most constituents change due to the heating process. Particularly, the heating promotes the development of 5-hydroxymethylfurfural (HMF) in sweet wines submitted to estufagem at higher temperatures. Moreover, estufagem provides the decrease of most amino acids, suggesting their involvement in the formation of the bouquet of these wines. Regarding the total polyphenol content and antioxidant potential of these wines they do not seem to be greatly affected by the heating step, however most monomeric polyphenols decrease during this process. The thermal processing of Madeira wines favours the development of the volatile composition, especially of volatiles usually reported as typical aromas of Madeira wines. Finally, it was demonstrated that the thermal degradation of sugars, especially of fructose, promotes the emergence of volatile compounds identified in baked wines.

Color measurement using a smartphone applied to Madeira wines

Computer vision is a field that uses techniques to acquire, process, analyze and understand images from the real world in order to produce numeric or symbolic information in the form of decisions [1]. This project aims to use computer vision to prepare an app to analyze a Madeira Wine and characterize it (identify its variety) by its color. Dry or sweet wines, young or old wines have a specific color. It uses techniques to compare histograms in order to analyze the images taken from a test sample inside a special container designed for this purpose. The color analysis from a wine sample using an image captured by a smartphone can be difficult. Many factors affect the captured image such as, light conditions, the background of the sample container due to the many positions the photo can be taken (different to capture facing a white wall or facing the floor for example). Using new technologies such as 3D printing it was possible to create a prototype that aims to control the effect of those external factors on the captured image. The results for this experiment are good indicators for future works. Although it’s necessary to do more tests, the first tests had a success rate of 80% to 90% of correct results. This report documents the development of this project and all the techniques and steps required to execute the tests.

Olfactory perception threshold assessment of volatile acidity in Madeira wines

Madeira wine is a fortified wine with impact in the Madeira Island’s economy. Similarly to other wines, its acidity should be well controlled in order to ensure Madeira wine quality, mostly the volatile acidity. Due to Madeira wine complex flavour, it is crucial to get a better knowledge about the volatile acidity impact in its features, namely determine the perception limit of acetic acid and ethyl acetate, as both are the main contributors for volatile acidity. Firstly, the olfactory perception threshold of volatile acidity was assessed by a trained and an untrained panel, using 5 and 10 years-old Sercial and Malvasia wines. Moreover, the current work also presents the evolution of organic acids, acetic acid and ethyl acetate during 540 days of ageing of Madeira wines (Malvasia, Bual, Verdelho and Sercial), comparing the same wines aged by both traditional ageing processes: canteiro and estufagem. Other wine samples, aged in wood casks (canteiro) for at least 5 years, were also evaluated. HS-SPME followed by GC-MS analysis was used to determine ethyl acetate concentration and IEC-HPLC-DAD was used for the organic acids determination, including acetic acid. The results indicated that acetic acid and ethyl acetate olfactory perception threshold depends essentially on wine’s age. Concerning acetic acid, the untrained panel was in average 5.45 g/L (5 years-old) and 6.22 g/L (10 years-old). Training the expert panel to recognize acetic acid odour, the values decreased for 1.44 g/L (5 years-old) and 1.87 g/L (10 years-old), but still remained higher than the established volatile acidity legal limits. Ethyl acetate threshold was similar for both panels (in average 327.97 mg/L). Both compounds tend to increase exponentially with age, being more evident in sweet wines. Organic acids in young Madeira wines depend mostly on the nature of grape varieties, but this difference is minimized with wine ageing.

Changes in volatile composition of Madeira wines during their oxidative ageing

The influence of the age in the volatile composition of Madeira wines made with Boal, Malvazia, Sercial and Verdelho varieties and aged in oak barrel during 1, 11 and 25 years old was been studied. For this purpose, the evolution of volatile compounds: higher alcohols, ethyl esters, fatty acids, furan compounds, enolic compounds, γ-lactones, dioxanes and dioxolanes, of the four most utilised varieties were determined using liquid–liquid extraction with dichloromeihane. Octan-3-ol was used as internal standard. The wines made with these varieties showed great differences in sugar content and small variations on pH and alcoholic degree. The results show that during ageing, the concentration of fatty acids ethyl esters, acetates and fatty acids decrease significantly contrarily to the great increase of ethyl esters of diprotic acids. There is a strong correlation between sotolon, 2-furfural, 5-methyl-2-furfural, 5-hydroxymethyl-2-furfural and 5-ethoxymethyl-2-furfural with wine ageing. These findings indicate that these compounds can be used as ageing wine markers. Among the molecules studied, sotolon [3-hydroxy-4,5-dimethyl-2(5H)-furanone] was one of the few molecules present in concentrations above the perception threshold in Madeira wines. 5-Eihoxymethyl-2-furfural formed from 5-hydroxymethyl-2-furfural and 2-furfural, derived from sugars, are also involved in the aroma of sweet fortified white wines aged in oxidative conditions. The sensory properties change significantly after long periods of conservation.

Analytical characterization of the aroma of Tinta Negra Mole red wine: Identification of the main odorants compounds

A method for the simultaneous determination of major and minor volatiles composition in different types (dry, medium dry, sweet and medium sweet) of a young Tinta Negra Mole (TNM) monovarietal red wine from 2003 harvest has been validated. Wine samples preparation includes a dichloromethane liquid–liquid extraction followed by concentration under a nitrogen atmosphere. The extracted fraction was analysed by gas chromatography–mass spectrometry and give quantitative information for more than 86 analytes whose concentration range from few μg l−1 to 259.1 mg l−1. The method enables high recovery of volatile compounds in wine good linearity with (r2) values higher than 0.980 and good sensitivity. The limits of detection range from 0.003 to 0.534 mg l−1 and limits of quantification from 0.009 to 1.170 mg l−1. The method allows satisfactory determination of more than 80 compounds in the TNM red wines. These wines are characterized by a high content of higher alcohols, ethyl esters, fatty acids and lactones. The levels of sulphur compounds in Tinta Negra Mole medium sweet wines are very low, but they have the highest concentration of carbonyl compounds. Quantitative analysis of the main odorants followed by the determination of aroma index allow us elucidate the aroma of these varieties. On the basis of their odour description and odour threshold, the most powerful odorants of Tinta Negra Mole wines were tentatively established.

Comprehensive two-dimensional gas chromatography with time-of-flight mass spectrometry combined with solid phase microextraction as a powerful tool for quantification of ethyl carbamate in fortified wines. The case study of Madeira wine

An analytical methodology based on headspace solid phase microextraction (HS-SPME) combined with comprehensive two-dimensional gas chromatography—time-of-flight mass spectrometry (GC × GC–ToFMS) was developed for the identification and quantification of the toxic contaminant ethyl carbamate (EC) directly in fortified wines. The method performance was assessed for dry/medium dry and sweet/medium sweet model wines, and for quantification purposes, calibration plots were performed for both matrices using the ion extraction chromatography (IEC) mode (m/z 62). Good linearity was obtained with a regression coefficient (r2) higher than 0.981. A good precision was attained (R.S.D. <20%) and low detection limits (LOD) were achieved for dry (4.31 μg/L) and sweet (2.75 μg/L) model wines. The quantification limits (LOQ) and recovery for dry wines were 14.38 μg/L and 88.6%, whereas for sweet wines were 9.16 μg/L and 99.4%, respectively. The higher performance was attainted with sweet model wine, as increasing of glucose content improves the volatile compound in headspace, and a better linearity, recovery and precision were achieved. The analytical methodology was applied to analyse 20 fortified Madeira wines including different types of wine (dry, medium dry, sweet, and medium sweet) obtained from several harvests in Madeira Island (Portugal). The EC levels ranged from 54.1 μg/L (medium dry) to 162.5 μg/L (medium sweet).