COMPARISON OF RNA EXTRACTION METHODS FOR Passiflora edulis SIMS LEAVES

ABSTRACT Functional genomic analyses require intact RNA; however, Passiflora edulis leaves are rich in secondary metabolites that interfere with RNA extraction primarily by promoting oxidative processes and by precipitating with nucleic acids. This study aimed to analyse three RNA extraction methods, Concert™ Plant RNA Reagent (Invitrogen, Carlsbad, CA, USA), TRIzol® Reagent (Invitrogen) and TRIzol® Reagent (Invitrogen)/ice -commercial products specifically designed to extract RNA, and to determine which method is the most effective for extracting RNA from the leaves of passion fruit plants. In contrast to the RNA extracted using the other 2 methods, the RNA extracted using TRIzol® Reagent (Invitrogen) did not have acceptable A260/A280 and A260/A230 ratios and did not have ideal concentrations. Agarose gel electrophoresis showed a strong DNA band for all of the Concert™ method extractions but not for the TRIzol® and TRIzol®/ice methods. The TRIzol® method resulted in smears during electrophoresis. Due to its low levels of DNA contamination, ideal A260/A280 and A260/A230 ratios and superior sample integrity, RNA from the TRIzol®/ice method was used for reverse transcription-polymerase chain reaction (RT-PCR), and the resulting amplicons were highly similar. We conclude that TRIzol®/ice is the preferred method for RNA extraction for P. edulis leaves.

Intraosseous migration of tendinous calcifications: cortical erosions, subcortical migration and extensive intramedullary diffusion, a SIMS series.

Calcium hydroxyapatite crystal deposition is a common disorder, which sometimes causes acute pain as calcifications dissolve and migrate into adjacent soft tissue. Intraosseous calcium penetration has also been described. We illustrate the appearance of these lesions using a series of 35 cases compiled by members of the French Society of Musculoskeletal Imaging (Société d'Imagerie Musculo-Squelettique, SIMS). The first group in our series (7 cases) involved calcification-related cortical erosions of the humeral and femoral diaphyses, in particular at the pectoralis major and gluteus maximus insertions. A second group (28 cases) involved the presence of calcium material in subcortical areas. The most common site was the greater tubercle of the humerus, accompanying a calcifying tendinopathy of the supraspinatus. In addition, an extensive intramedullary diffusion of calcium deposits was observed in four of these cases, associated with cortical erosion in one case and subcortical lesions in three cases. Cortical erosions and intraosseous migration of calcifications associated with calcific tendinitis may be confused with neoplasm or infection. It is important to recognize atypical presentations of hydroxyapatite deposition to avoid unnecessary investigation or surgery.

Histopathological features of infections caused by Fusarium oxysporum and F. solani in purple passionfruit plants (Passiflora edulis Sims)

The purple passionfruit plant, Passifloraedulis Sims, ranks second in fruit exportation in Colombia, and its main destination is the European market. However, its production is affected by several diseases, including fusariosis. This paper presents the histopathological features of different tissues affected by the pathogens Fusarium oxysporum and Fusarium solani. Both microorganisms produce similar responses on the plant: colonization of xylem vessels by hyphae and microconidia, hypertrophy and hyperplasia of the cambium, xylem and phloem; destruction of xylem fibers and amyloplasts in parenchymatous cells; and production of gels by the plant. However, there are differences in the colonization mechanism, F. solani penetrates and is concentrated especially at the collar zone, while F. oxysporum penetrates the roots and moves through the vascular system to colonize the plant.

Virtuaalilähiöstä draaman näyttämöksi. The Sims visuaalisena ja kerronnallisena työkaluna

Tutkimuksen tarkoituksena oli tarkastella ilmiötä, jossa digitaalista peliä ei enää vain pelata, vaan sitä käytetään uusilla innovatiivisilla tavoilla kirjallisten ja visuaalisten tuotosten tekemiseen. Tarkastelun pääkohteena vuonna 2004 julkaistu peli The Sims 2, mutta tutkimus on sovellettavissa myös pelisarjan muihin osiin. Tutkimuksessa tartuttiin yhteen tiettyyn luovan adaptaation muotoon, joista peliyhteisöissä käytetään nimitystä Sims-tarinat. Nämä ovat pelaajien tekemiä narratiivisia ja graafisia esityksiä, joiden toteuttamiseksi peli sovelletaan työkaluksi ja käytetään visuaalisena alustana. Ilmiö on lähtenyt yhteisön tavasta jakaa pelitilaansa kuvakaappausten kautta. Tutkimuksessa tarkasteltiin erityisesti, millaisia nämä tuotokset ovat, miksi niitä tehdään ja millaisessa yhteydessä ne ovat itse pelaamiseen. Aineistona käytettiin ensisijaisesti havainnointia ja verkkokyselyä. Tutkimuksen metodina oli laadullinen sisällönanalyysi, jonka kautta muodostettiin erilaisia tyypittelyjä ilmiöstä. Tarkasteltavana olivat pääasiallisesti sivustot Mod The Sims sekä Livejournalin Sims storytellers, joissa havainnointia toteutettiin satunnaisotannalla. Samoilla sivustoilla julkaistiin myös verkkokysely. Tarinoissa yhdistyy pelistä otetut kuvakaappaukset sekä teksti. Niiden draamallinen sisältö on vaihteleva ja voi käsitellä kaikkia yleisesti tunnettuja kerronnan genrejä. Tarinat voivat olla kiinteä osa itse pelaamista tai siitä irrallisempia. Joillekin peli on yksinomaan alusta, jolla toteuttaa tarinoita, mutta useimmille ne esiintyvät myös pelikokemusta rikastavana ja muokkaavana harrastuksena. Niissä lavastetaan usein tilanteita, jotka eivät itse pelatessa ole mahdollisia, jolloin myös peli saa uusia merkityksiä pelimaailman ulkopuolelta. Tarinat vaikuttavat peliin ja pelin kulku vaikuttaa tarinoihin. Ne ovat esimerkki emergentistä pelattavuudesta ja kuluttajien tavasta muokata kulttuurituotteet omiin tarpeisiin soveltuvaksi, mikä luo uusia pelaamisen muotoja. Sims-tarinoita kirjoittavat etupäässä aikuiset naiset, jotka ovat halunneet yhdistää mieluisat harrastukset. He arvottivat tarinoita pääosin samoin perustein kuin muitakin kirjallisia tuotoksia, mutta uusia elementtejä olivat niiden visuaalisuus ja yhteisöllisyys. Useimmiten motivaatio harrastukseen lähti peliyhteisön esimerkistä. Toinen yleinen teema oli se, kuinka pelaajat pelatessaan alkoivat automaattisesti luoda omaa narratiivisuutta ja kokivat tarvetta tuoda nämä ideat konkreettisen tarinan muotoon, jonka jakaa yhteisössä. Tutkimus osoitti, että pelit sallivat harrastajien toimia omaehtoisina kulttuurintuottajina ja synnyttää uusia innovatiivisia pelaamisen muotoja. Jatkotutkimuksessa on huomionarvoista kuinka nykypäivän mediakulutus ja -tuotanto integroituvat yhä tiiviimmin toisiinsa. Näiden uusien ilmiöiden tutkimus on siten tärkeää ja edellyttää moninäkökulmaista tutkimusotetta.

Topochemistry of physical and chemical pretreatments of biomass as investigated by FE-SEM, XPS and ToF-SIMS

Surface chemistry is of great importance in plant biomass engineering and applications. The surface chemical composition of biomass which includes lignin, carbohydrates and extractives influences its interactions with chemical agents, such as pulp processing/papermaking chemicals, or enzymes for different purposes. In this thesis, the changes in the surface chemical composition of lignocellulosic biomass after physical modification for the improvement of resulting paper properties and chemical treatment for the enhancement of enzymatic hydrolysis were investigated. Low consistency (LC) refining was used as physical treatment of bleached softwood and hardwood pulp samples, and the surface chemistry of refined samples was investigated. The refined pulp was analysed as whole pulp while the fines-free fibre samples were characterized separately. The fines produced in LCrefining contributed to an enlarged surface specific area as well as the change of surface coverage by lignin and extractives, as investigated by X-ray photoelectron spectroscopy (XPS). The surface coverage by lignin of the whole pulp decreased after refining while the surface coverage by extractives increased both for pine and eucalyptus. In the case of pine, the removal of fines resulted in reduction of the surface coverage by extractives, while the surface coverage by lignin increased on fibre sample (without fines). In the case of eucalyptus, the surface coverage by lignin of fibre samples decreased after the removal of fines. In addition, the surface distribution of carbohydrates, lignin and extractives of pine and eucalyptus samples was determined by Time-of-Flight Secondary Ion Mass Spectrometry (ToF-SIMS). LC-refining increased the amounts of pentose, hexose and extractives on the surface of pine samples. ToF-SIMS also gave clear evidence about xylan deposition and reduction of surface lignin distribution on the fibre of eucalyptus. However, the changes in the surface chemical composition during the physical treatment has led to an increase in the adsorption of fluorescent whitening agents (FWAs) on fibres due to a combination of electro-static forces, specific surface area of fibres and hydrophobic interactions. Various physicochemical pretreatments were conducted on wood and non-wood biomass for enhancing enzymatic hydrolysis of polysaccharides, and the surface chemistry of the pretreated and enzymatically hydrolysed samples was investigated by field emission scanning electron microscopy (FE-SEM), XPS and ToF-SIMS. A hydrotrope was used as a relatively novel pretreatment technology both in the case of wood and non-wood biomass. For comparison, ionic liquid and hydrothermal pretreatments were applied on softwood and hardwood as well. Thus, XPS analysis showed that the surface lignin was more efficiently removed by hydrotropic pretreatment compared to ionic liquid or hydrothermal pretreatments. SEM analysis also found that already at room temperature the ionic liquid pretreatments were more effective in swelling the fibres compared with hydrotropic pretreatment at elevated temperatures. The enzymatic hydrolysis yield of hardwood was enhanced due to the decrease in surface coverage of lignin, which was induced by hydrotropic treatment. However, hydrotropic pretreatment was not appropriate for softwood because of the predominance of guaiacyl lignin structure in this material. In addition, the reduction of surface lignin and xylan during pretreatment and subsequent increase in cellulose hydrolysis by enzyme could be observed from ToF-SIMS results. The characterisation of the non-wood biomass (e.g. sugarcane bagasse and common reed) treated by hydrotropic method, alkaline and alkaline hydrogen peroxide pretreatments were carried out by XPS and ToF-SIMS. According to the results, the action for the removal of the surface lignin of non-wood biomass by hydrotropic pretreatment was more significant compared to alkaline and alkaline hydrogen peroxide pretreatments, although a higher total amount of lignin could be removed by alkaline and alkaline hydrogen peroxide pretreatment. Furthermore, xylan could be remarkably more efficiently removed by hydrotropic method. Therefore, the glucan yield achieved from hydrotropic treated sample was higher than that from samples treated with alkaline or alkaline hydrogen peroxide. Through the use of ToF-SIMS, the distribution and localization of lignin and carbohydrates on the surface of ignocelluloses during pretreatment and enzymatic hydrolysis could be detected, and xylan degradation during enzymatic hydrolysis could also be assessed. Thus, based on the results from XPS and ToF-SIMS, the mechanism of the hydrotropic pretreatment in improving the accessibility of enzymes to fibre and further ameliorating of the enzymatic saccharification could be better elucidated.

Uso da cor da casca como indicador de qualidade do maracujá amarelo (Passiflora edulis Sims. f. flavicarpa Deg.) destinado à industrialização

As características físico-químicas do maracujá amarelo em três estádios de cor de casca (1/3 amarelo, 2/3 amarelo e inteiro amarelo), em quatro épocas da safra/99 foram avaliadas, visando estabelecer o ponto de colheita de melhor qualidade da fruta para a industrialização. Foram determinados o tamanho dos frutos, a cor da casca e da polpa, o rendimento de extração da polpa, o teor de sólidos solúveis totais, o pH, a acidez total, o ratio e o teor de vitamina C. Foi empregado o delineamento experimental inteiramente casualizado, com 8 repetições, 5 frutos por parcela e 3 tratamentos. Os resultados mostraram que o rendimento de polpa não apresentou diferença significativa entre os tratamentos e entre as colheitas. Com relação à cor da polpa, houve diferença significativa entre os três estádios apenas na 1ª colheita. Os frutos de casca 1/3 amarela apresentaram teor de sólidos solúveis significativamente inferior aos demais apenas na 3ª e 4ª colheitas, mas o maior valor médio ocorreu na 4ª colheita. O teor de acidez total dos frutos de casca inteira amarela foi significativamente inferior àqueles das colheitas 2, 3 e 4. Os valores mais elevados de vitamina C foram obtidos na 1ª colheita e, os frutos de casca 1/3 amarela apresentaram teores significativamente inferiores nas colheitas 1 e 2. De maneira geral, os resultados indicaram que, embora tenham ocorrido algumas diferenças nas características físico-químicas dos frutos nos diferentes estádios de cor de casca, frutos colhidos em todos os estádios de cor de casca estudados apresentaram-se adequados à industrialização.

Conservação de sementes de maracujá-amarelo (Passiflora edulis Sims f. flavicarpa Deg.)

Empregando variações no grau de umidade das sementes e na temperatura do ambiente, a pesquisa objetivou estudar o comportamento das sementes de maracujá-amarelo (Passiflora edulis Sims f. flavicarpa Deg.) durante o armazenamento. Para tanto, as sementes, uma vez extraídas dos frutos, receberam turbilhonamento hídrico e foram atritadas sobre peneira de arame trançado, durante a lavagem em água corrente, para a retirada parcial da mucilagem. Após a eliminação da água aderida externamente às sementes, através de secagem à sombra, foi obtido o tratamento com 26% de H2O; as sementes restantes foram submetidas à secagem em estufa para a obtenção dos demais tratamentos (20, 14, 10 e 8% de H2O). As sementes, embaladas em sacos de polietileno, foram armazenadas em ambiente com temperatura controlada de 10 e 20°C; no início do armazenamento, e após 35, 70, 105, 140, 175, 210, 245, 280 e 315 dias, as sementes foram submetidas às avaliações da qualidade. A conservação das sementes, considerando os intervalos estudados, é favorecida pela combinação do grau de umidade de 10% com a temperatura de 20°C. O desempenho fisiológico, por outro lado, apresenta variações temporais sugestivas de efeitos de dormência. Entretanto, a recorrência e a aleatoriedade observadas, sem interferências ambientais evidentes, não encontram respaldo na fenomenologia atribuída à dormência pela literatura especializada; dessa forma, o tema demanda investigações específicas.

Secagem e armazenamento de sementes de maracujá amarelo (Passiflora edulis Sims f. flavicarpa Degener)

Este trabalho foi desenvolvido com os objetivos de avaliar os efeitos imediato e latente da temperatura de secagem e de dois tipos de embalagem sobre a qualidade fisiológica de sementes de maracujá-amarelo após três e seis meses de armazenamento. Empregaram-se quatro níveis de temperatura do ar de secagem (ambiente à sombra, 30, 35 e 40 ºC), um nível de fluxo de ar, 48 m³ min-1 m-2, e teores de água inicial e final de cerca de 35 e 10 % b.u., respectivamente. Os tratamentos de secagem foram realizados em protótipo de secador de leito fixo, em camada delgada, com fluxo de ar ascendente. Avaliou-se a qualidade fisiológica por meio de testes de germinação e vigor. Depois da secagem, as sementes foram embaladas e armazenadas em câmara do tipo B.O.D. a 15±1 ºC. As temperaturas utilizadas na secagem das sementes não influenciam a germinação. O armazenamento por três e seis meses não afetou a qualidade fisiológica das sementes. A secagem das sementes de mamão pode ser realizada à sombra ou em temperaturas de 30 a 40 ºC sem afetar a qualidade fisiológica das mesmas.

Videojuegos y Redes Sociales : el proceso de identidad en Los Sims 3.

Monográfico con el título: 'Sobre vídeojuegos y aprendizaje'. Resumen basado en el de la publicación

Eyes, amygdala, and other models of face processing: Questions for the SIMS model

This commentary raises general questions about the parsimony and generalizability of the SIMS model, before interrogating the specific roles that the amygdala and eye contact play in it. Additionally, this situates the SIMS model alongside another model of facial expression processing, with a view to incorporating individual differences in emotion perception.

SEM, EPR and ToF-SIMS analyses applied to unravel the technology employed for pottery-making by pre-colonial Indian tribes from Pantanal, Brazil

The present work aims at elucidating the technology applied in the fabrication of ceramic objects by the ancient ceramists that inhabited the western border of Pantanal, Mato-Grosso do Sul, with the help of a multidisciplinary approach making use of chemical and physical methods of analysis. The potshards under study show the presence of different types of additives, as determined by scanning electron microscopy (SEM) and time of flight secondary ion mass spectrometry (ToF-SIMS). The dispersion of the additives within the ceramic matrix was also addressed by SEM, which shed light on the mounting technique used by the potters to assemble the ceramic vessels. Moreover, the tensile strength conferred to the pottery by the use of a specific type of additive was evaluated by applying a mechanical test. These results were correlated with the firing temperature of the potshards, determined by means of electron paramagnetic resonance (EPR). (C) 2010 Elsevier Ltd. All rights reserved.

Investigation of the drying process of linseed oil using FTIR and ToF-SIMS

The drying process of linseed oil, oxidized at 80 oC, has been investigated with rheology measurements, Fourier transformation infrared spectroscopy (FTIR), and time of flight secondary ion mass spectrometry (ToF-SIMS). The drying process can be divided into three main steps: initiation, propagation and termination. ToF-SIMS spectra show that the oxidation is initiated at the linolenic (three double bonds) and linoleic fatty acids (two double bonds). ToF-SIMS spectra reveal peaks that can be assigned to ketones, alcohols and hydroperoxides. In this article it is shown that FTIR in combination with ToF-SIMS are well suited tools for investigations of various fatty acid components and reaction products of linseed oil.

Propriedades antioxidantes de extratos de Passiflora alata Dryander e de Passiflora edulis Sims

O presente estudo investigou as atividades antioxidantes in vitro, ex vivo e in vivo dos extratos de folhas Passiflora alata e Passiflora edulis, plantas usadas na medicina popular e ricas em polifenóis, compostos com reconhecida atividade antioxidante. No modelo experimental in vitro, ambos os extratos demonstraram atividade antioxidante e proteção contra dano protéico induzido por glicose. Fatias de fígado de ratos foram utilizadas como modelo ex vivo. Tanto o extrato de P. alata quanto o extrato de P. edulis protegeram de forma significativa o dano protéico e a morte celular induzidos por FeSO4. Como a P. alata é uma droga oficial da Farmacopéia Brasileira, os efeitos antioxidantes deste extrato foram investigados in vivo. Ratos machos Wistar receberam tratamento intragástrico de extrato de folhas de P. alata (1 e 5 mg/kg), trolox (0,18 mg/kg) ou água (controle) durante 30 dias, seguido de uma dose de CCl4 (3 ml/kg, i.p.) no 30º dia. O dano hepático e os efeitos antioxidantes do pré-tratamento com extrato de P. alata foram avaliados em vários órgãos. Quando comparados ao grupo controle, os ratos pré-tratados com o extrato demonstraram dano hepático menor, evidenciado por um grau menor de necrose, níveis menores de lipoperoxidação e maior atividade das enzimas superóxido dismutase e catalase. Adicionalmente, níveis menores de lipoperoxidação cardíaca foram observados com o pré-tratamento de P. alata (5mg/kg). Os resultados obtidos neste estudo indicam que os extratos de Passiflora são fontes potenciais de antioxidantes naturais. Estudos adicionais investigando o papel de compostos isolados destes extratos em patologias humanas onde o estresse oxidativo está envolvido são necessários.

Uso da cor da casca como indicador de qualidade do maracujá amarelo (Passiflora edulis Sims. f. flavicarpa Deg.) destinado à industrialização

As características físico-químicas do maracujá amarelo em três estádios de cor de casca (1/3 amarelo, 2/3 amarelo e inteiro amarelo), em quatro épocas da safra/99 foram avaliadas, visando estabelecer o ponto de colheita de melhor qualidade da fruta para a industrialização. Foram determinados o tamanho dos frutos, a cor da casca e da polpa, o rendimento de extração da polpa, o teor de sólidos solúveis totais, o pH, a acidez total, o ratio e o teor de vitamina C. Foi empregado o delineamento experimental inteiramente casualizado, com 8 repetições, 5 frutos por parcela e 3 tratamentos. Os resultados mostraram que o rendimento de polpa não apresentou diferença significativa entre os tratamentos e entre as colheitas. Com relação à cor da polpa, houve diferença significativa entre os três estádios apenas na 1ª colheita. Os frutos de casca 1/3 amarela apresentaram teor de sólidos solúveis significativamente inferior aos demais apenas na 3ª e 4ª colheitas, mas o maior valor médio ocorreu na 4ª colheita. O teor de acidez total dos frutos de casca inteira amarela foi significativamente inferior àqueles das colheitas 2, 3 e 4. Os valores mais elevados de vitamina C foram obtidos na 1ª colheita e, os frutos de casca 1/3 amarela apresentaram teores significativamente inferiores nas colheitas 1 e 2. de maneira geral, os resultados indicaram que, embora tenham ocorrido algumas diferenças nas características físico-químicas dos frutos nos diferentes estádios de cor de casca, frutos colhidos em todos os estádios de cor de casca estudados apresentaram-se adequados à industrialização.