CD4 T-Cell Cytokine Response to Mite Recombinant Tropomyosin in Mite, Snail and Shrimp Allergic Patients

This work is a preliminary study on immune cell function, evaluating possible tropomyosin-triggering in a snail-mite-shrimp cross-reaction

Selected cardoon (Cynara cardunculus L.) genotypes suitable for PDO cheeses in Mediterranean regions

Cardoon flower extract is a traditional and exclusive rennet used for some PDO cheeses in several Mediterranean regions, due to its extremely high concentration in cardosins. In this preliminary study, six individual cardoon genotypes (1M – 6M) were selected because they revealed a wide and consistent diversity of total and specific cardosin concentrations in flowers. During three growing seasons, the stability of 12 biochemical characteristics of flower extracts and 26 plant morphological descriptors was confirmed. Surprisingly, the cardosin profiles of each genotype, based on four main groups A0, A1, A and B, were stable during the annual flower harvesting period and over all three years using ion-exchange chromatography and native-PAGE electrophoresis. This knowledge will allow an improvement in the quality and standardization of cardosin profiles from cardoon flowers used for cheese production and other innovative applications. The results obtained are promising for the development of a plant breeding program based on biochemical and morphological characteristics in order to obtain the most adapted plant architecture for combined purposes related to specific cardosins composition, flower and plant biomass production, and ease of harvesting.info:eu-repo/semantics/acceptedVersio

Specificity and kinetics of the milk-clotting enzyme from cardoon (Cynara cardunculus L.) toward bovine .kappa.-casein

The action of Cynara cardunculus L. protease on whole bovine K-casein, over a 3-h period at pH 6.4, was investigated. RpHPLC of the 3% trichloroacetic acid (TCA)-solublefraction of the K-casein digestion mixture showed three peptide peaks, which were identified by amino acid analysis and N-terminal analysis as the 106-169 fragment [caseinomacropeptide (CMP)]. Upon selective precipitation with 12% TCA, one glycosylated and two nonglycosylated forms of CMP were distinguished. Analysis of the whole digestion mixture showed no additional peptides. The kinetics of hydrolysis of the PhelO5- Met106 bond was studied by spectrofluorometry, using fluorescein isothiocyanate-labeled K-casein (FTC- K-casein). The values obtained for kat, k, and k were 1.04 s-l, 0.16 pM, and 6.5 pM-l s-l, respectively. The proteolytic coefficient is of the same order of magnitude as those obtained for other milk-clotting enzymes, but the k, is significantly lower, which reflects the higher affinity of Cynara protease to K-casei

Cardosins A and B, two new enzymes available for peptide synthesis

Two new aspartic proteases, Cardosins A and B, with a high specificity toward bonds between hydrophobic amino acids were isolated from the flowers of the cardoon, Cynara cardunculus L., and recently characterised [C.J. Faro, A.G.J. Moir, E. Pires, Biotech. Lett., 14 (1992) 841.]; [P. Veríssimo, C. Faro, A.J.G. Moir, Y. Lin, J. Tang, E. Pires, Eur. J. Biochem., 235 (1996) 762.]. Cardosins were shown to be stable in aqueous-organic biphasic systems [M. Barros, M.G.V. Carvalho, F.A. Garcia, E. Pires, Biotech. Lett. 14 (1992) 174.]. In this work, we have investigated peptide bond specificity of Cardosin A and Cardosin B in what concerns the amino acids in P'1 position. The results were compared with pepsin under the same conditions. Information about secondary specificity of Cardosin A and B was also investigated by tripeptide synthesis. The condensation reactions were carried out in aqueous-organic biphasic systems of n-hexane/ethyl acetate and sodium phosphate buffer. The reaction products were isolated by RP-HPLC and identified by amino acid analysis and eventually by M.S. The results in the synthesis of dipeptides showed that Cardosin A and B have similar P'1 position preference. The production of tripeptides by condensation of CBZ·Val·Phe with Phe·OMe, Met·OMe and Val·OMe reveals that the addition of Val in the P2 position modifies the Cardosins' preferences concerning the amino acid in P'1 position.http://www.sciencedirect.com/science/article/B6TGN-3V5RY4G-2B/1/1e2fd83ec2ea4e623929f202f28a6d5

Chemical treatment to unclogg dripper irrigation systems due to biological problems

Muitos produtores usam gotejadores para irrigação localizada em produção de flores no campo e em ambientes protegidos. Um problema freqüente neste tipo de irrigação é o entupimento dos gotejadores, que está diretamente relacionado à qualidade da água e à deficiência de filtração do sistema. Este trabalho teve como objetivo fazer uma investigação experimental para avaliar a eficiência da utilização de ácido nítrico e de hipoclorito de sódio na desobstrução de gotejadores entupidos, devido à utilização de águas com elevado teor de matéria de origem biológica. A avaliação foi realizada em seis estufas de 4216 m², sendo que cada estufa tem dois setores, composto de dez vãos ou linhas em cada setor, totalizando 12 setores de um sistema de irrigação por gotejamento, em propriedade produtora de rosas (Rosa sp.), sob estufa plástica, no município de Holambra, SP. Análises química, física e de contagem bacteriológica da água foram realizadas nas três fontes hídricas que abasteciam o sistema de irrigação, para verificar quais os fatores que estavam causando risco de entupimento. Foram feitas avaliações da uniformidade de distribuição de água em todos os setores antes e depois do tratamento químico com o intuito de avaliar a sua eficiência. Ocorreu melhora na uniformidade de distribuição de água e diminuição no coeficiente de variação de vazão (CV) dos gotejadores em todos os setores. Houve boa correlação entre o CV e o índice de uniformidade adotado nesta pesquisa. Portanto este é um excelente método para ser utilizado na desobstrução de gotejadores entupimentos devido a problemas de origem biológica.Many producers use drippers for trickle irrigation systems for flower production in the field and in protected environments. A frequent problem in this type of irrigation system is the clogging of drippers, which is directly related to water quality and filtering system efficiency. The objective of this study was to conduct an experimental investigation to evaluate the efficiency of using nitric acid and sodium hypochlorite to unclogg drippers due to the use of water with high algae content. The evaluation was conducted in six, 4216 m² greenhouses, each with two sectors comprised of ten spaces or lines, totaling 12 sectors of a dripper irrigation system in a rose producing property of Holambra, State of SP, Brazil. Chemical and physical analyses and the bacteriological count in water were carried out in the three water sources that supply the irrigation system to check the factors causing the clogging. Evaluations were carried out on water distribution uniformity in all sectors before and after chemical treatment in order to evaluate efficiency. The treatment improved water distribution uniformity and a lead to a reduction in the coefficient of variation (CV) for dripper flow in all sectors. There was a good correlation between CV and the water distribution uniformity index. Therefore, this is an excellent method to be used to unclogg drippers due to biological problems

Effect of gelation temperature on the properties of skim milk gels made from plant coagulants and chymosin

Reconstituted skim milk was gelled at 25-40°C with the plant-origin coagulants from Cynara cardunculus L. or Cynara humilis L. or with fermentation-produced chymosin. Gel formation and ageing were monitored by low amplitude oscillatory rheology and confocal scanning laser microscopy. Arrhenius plots for the rate of milk gelation were also determined. Plant coagulants had shorter gelation time (tg) at 25°C, 35°C and 40°C, and higher initial rate of increase in G' values at all temperatures tested. The firmest gels at long ageing times were produced by chymosin at 30°C and 32°C. At a gelation temperature of 25°C, the differences in rheological and microstructural characteristics between plant coagulants and chymosin were considerable; plant coagulants had shorter tg and higher G' values. For the lowest gelation temperatures, plant coagulants had smaller activation energy values for gelation. Most of the gelation results were similar between plant coagulants, but some differences were found in the values of tg, the rate of increase in G' and loss tangent parameter. The characteristics of gels produced with plant coagulants were influenced less by the changes in temperature compared with chymosin-produced gels, which may be an important consideration in using plant-origin coagulants in the production of cheeses with a wider range of gelation temperatures.http://www.sciencedirect.com/science/article/B6T7C-493HNG1-1/1/35f20b14e49b2922b16639bac3576d1

Molecular analysis of the interaction between cardosin A and phospholipase Dα

Cardosin A is an RGD-containing aspartic proteinase from the stigmatic papillae of Cynara cardunculus L. A putative cardosin A-binding protein has previously been isolated from pollen suggesting its potential involvement in pollen2013pistil interaction [Faro C, Ramalho-Santos M, Vieira M, Mendes A, Simões I, Andrade R, Verissimo P, Lin X, Tang J & Pires E (1999) J Biol Chem274, 28724201328729]. Here we report the identification of phospholipase D03B1 as a cardosin A-binding protein. The interaction was confirmed by coimmunoprecipitation studies and pull-down assays. To investigate the structural and molecular determinants involved in the interaction, pull-down assays with cardosin A and various glutathione S-transferase-fused phospholipase D03B1 constructs were performed. Results revealed that the C2 domain of phospholipase D03B1 contains the cardosin A-binding activity. Further assays with mutated recombinant forms of cardosin A showed that the RGD motif as well as the unprecedented KGE motif, which is structurally and charge-wise very similar to RGD, are indispensable for the interaction. Taken together our results indicate that the C2 domain of plant phospholipase D03B1 can act as a cardosin A-binding domain and suggest that plant C2 domains may have an additional role as RGD/KGE-recognition domains

Least limiting water range in the evaluation of continuous and short-duration grazing systems

A qualidade física do solo sob sistemas de pastejo contínuo e rotacionado foi avaliada pelo Intervalo Hídrico Ótimo (IHO). A amostragem foi realizada na área experimental da Embrapa Gado de Corte. Foi estudado um Latossolo Vermelho distrófico. Foram avaliados quatro piquetes, sendo dois no sistema de pastejo contínuo e dois no sistema de pastejo rotacionado. No sistema de pastejo contínuo, implementado com a espécie Brachiaria decumbens cv. Basilisk, foram retiradas 30 amostras por piquete, não tendo um dos piquetes recebido adubação de manutenção (Cs), enquanto o outro havia recebido adubação bianual de manutenção (Cc). No sistema de pastejo rotacionado, implementado com a espécie Panicum maximum cv. Tanzânia, foram retiradas 30 amostras por piquete; em um dos piquetes, o resíduo pós-pastejo era mantido entre 2,0 e 2,5 t ha-1 de matéria seca total (MST) (R1), enquanto no outro era mantido entre 3,0 e 3,5 t ha-1 MST (R2). As amostras foram submetidas a um gradiente de tensão de água e, posteriormente, utilizadas nas determinações da densidade do solo (Ds), resistência do solo à penetração (RP), umidade volumétrica (qv) e do IHO. O sistema de pastejo rotacionado apresentou piores condições físicas do solo para o crescimento vegetal, avaliadas pelo critério do IHO. Os maiores valores de Ds e menor IHO foram observados no R1, o que foi atribuído às taxas de lotação mais elevadas aplicadas neste piquete. __________________________________________________________________________________________ ABSTRACTSoil physical quality in continuous and short-duration rotational cattle grazing systems was evaluated using the Least Limiting Water Range (LLWR) approach. Soil samples were collected on an experimental site at the Embrapa – Beef Cattle Research Center (Campo Grande, MS, Brazil). The studied soil was a Typic Acrudox. Four sampling sites were selected: two under a short-duration continuous grazing system and two under intensive short-duration rotational grazing system. Thirty soil cores were collected in each site under the continuous grazing system, planted with Brachiaria decumbens cv. Basilisk grass. One of the continuous grazing sites had received biannual fertilization (Cc) while the other had not been fertilized (Cs). Thirty soil cores were also collected in each site under the intensive rotational grazing system, planted with Panicum maximum cv. Tanzania grass. In the intensive rotational grazing system the two sites differed in relation to the post-graze residue level left. In one site (R1) it varied from 2.0 to 2.5 t total dry matter (TDM) ha-1 and in the other R2 it varied from 3.0 to 3.5 t TDM ha-1. The cores were subjected to a soil water suction gradient, and then used for soil bulk density (Db), soil resistance to penetration (SR), volumetric water content (qv), and LLWR determinations. The intensive short-duration rotational grazing system presented the worse physical soil conditions for plant growth, as evaluated by the LLWR. The highest Db and the lowest LLWR were found in R1, which could be a consequence of the heavier stocking rates used on this site

Performance of non woven synthetic fabric and disc filters for fertirrigation water treatment

Um dos fatores que elevam os custos de operação e manutenção dos sistemas de irrigação localizada e, em certos casos, inviabiliza a utilização desse método, é a obstrução de emissores pela presença de partículas sólidas na água de irrigação. O presente trabalho teve por objetivo principal estimar, através de ensaios experimentais de campo, o desempenho de um filtro de disco (130 microns) com outro de manta sintética não tecida na remoção de impurezas de origens físicas, químicas e biológicas presentes na água de irrigação que promovem o entupimento dos emissores, quando se utiliza a fertirrigação. Os critérios para avaliação das impurezas presentes na água de irrigação foram baseados nos estudos realizados em sistemas de irrigação localizada. Especificamente foram analisados os seguintes parâmetros físico-químicos e biológicos da água afluente e efluente dos filtros: pH, turbidez, sólidos suspensos totais, salinidade, ferro total, sulfetos, manganês, algas e bactérias. Os fatores químicos que apresentaram médio risco de obstrução aos emissores foram pH, concentração de ferro total e sulfetos. Todos os demais parâmetros de qualidade de água analisados resultaram em valores que não apresentam risco de entupimento aos gotejadores. Procurou-se também fazer o acompanhamento da evolução da perda de carga em função do volume filtrado para os filtros, para realizar uma análise comparativa. No filtro de manta sintética não tecida, a evolução da perda de carga foi mais acentuada e mais rápida em relação ao de disco, por causa da sua maior eficiência na remoção de algas e bactérias.Obstruction of emitters caused by the presence of solid particles in the water raises the cost and maintenance of trickle irrigation systems, and might compromise their utilization. This research was performed through test fields, with the objective of evaluating the performance of a disc filter (130 microns) and a non woven synthetic fabric filter on the removal of physical, chemical and biological impurities from the irrigation water, which may cause emitter obstruction during fertirrigation processes. The evaluation criteria of the impurities present in the irrigation water, were based on studies performed on trickle irrigation systems. Specifically, physical, chemical and biological parameters analyzed in the inflow and outflow water from both filters were: pH, turbidity, total suspended solids, salinity, total iron, sulphites, manganese, algae and bacteria. Results pointed to chemical factors, which presented medium clogging risks to the emitters; those factors were: pH, total iron and sulphite concentration. All the other parameters of water quality did not present clogging risks. A comparative analysis of head loss evolution, according to the filtered volume, was also atempted Non woven synthetic fabric filter presented a higher evolution when compared to the disc filter