Effect of storage time and conditions on the seed coat colour of Australian adzuki beans

Two varieties of adzuki beans (Vigna angularis), Bloodwood and Erimo, were stored at temperatures of 10, 20 or 30degreesC, and relative humidities (RH) 40 or 65%, and samples were analysed at 0, 1.5, 3 and 6 months. Storage at 30degreesC for > 1.5 months caused a significant decrease in the a(star) and b(star) colour values and darkening of the seed coat. Beans stored at 65% RH had lower L-star but higher a(star) and b(star) colour values than those stored at 40% RH. Bloodwood and Erimo samples showed similar trends in colour during storage. The best storage conditions for the preservation of the adzuki colour were 10degreesC and 65% RH. The Australian beans had lower L-star, a(star) and b(star) colour values than Japanese Erimo-shouzu beans and storage increased the difference.

Isolation and characterization of a cone snail protease with homology to CRISP proteins of the pathogenesis-related protein superfamily

The pathogenesis-related (PR) protein superfamily is widely distributed in the animal, plant, and fungal kingdoms and is implicated in human brain tumor growth and plant pathogenesis. The precise biological activity of PR proteins, however, has remained elusive. Here we report the characterization, cloning and structural homology modeling of Tex31 from the venom duct of Conus textile. Tex31 was isolated to >95% purity by activity-guided fractionation using a para-nitroanilide substrate based on the putative cleavage site residues found in the propeptide precursor of conotoxin TxVIA. Tex31 requires four residues including a leucine N-terminal of the cleavage site for efficient substrate processing. The sequence of Tex31 was determined using two degenerate PCR primers designed from N-terminal and tryptic digest Edman sequences. A BLAST search revealed that Tex31 was a member of the PR protein superfamily and most closely related to the CRISP family of mammalian proteins that have a cysteine-rich C-terminal tail. A homology model constructed from two PR proteins revealed that the likely catalytic residues in Tex31 fall within a structurally conserved domain found in PR proteins. Thus, it is possible that other PR proteins may also be substrate-specific proteases.

Structural characterization of respiratory syncytial virus fusion inhibitor escape mutants: homology model of the F protein and a syncytium formation assay

Respiratory syncytial virus (RSV) is a ubiquitous human pathogen and the leading cause of lower respiratory tract infections in infants. Infection of cells and subsequent formation of syncytia occur through membrane fusion mediated by the RSV fusion protein (RSV-F). A novel in vitro assay of recombinant RSV-F function has been devised and used to characterize a number of escape mutants for three known inhibitors of RSV-F that have been isolated. Homology modeling of the RSV-F structure has been carried out on the basis of a chimera derived from the crystal structures of the RSV-F core and a fragment from the orthologous fusion protein from Newcastle disease virus (NDV). The structure correlates well with the appearance of RSV-F in electron micrographs, and the residues identified as contributing to specific binding sites for several monoclonal antibodies are arranged in appropriate solvent-accessible clusters. The positions of the characterized resistance mutants in the model structure identify two promising regions for the design of fusion inhibitors. (C) 2003 Elsevier Science (USA). All rights reserved.

Generation of aldehyde-derived protein modifications in ethanol-exposed heart

Background: Although excessive ethanol consumption is known to lead to a variety of adverse effects in the heart, the molecular mechanisms of such effects have remained poorly defined. We hypothesized that posttranslational covalent binding of reactive molecular species to proteins occurs in the heart in response to acute ethanol exposure. Methods: The generation of protein adducts with several aldehydic species was examined by using monospecific antibodies against adducts with malondialdehyde (MDA), acetaldehyde (AA), MDA-AA hybrids, and hydroxyethyl radicals. Specimens of heart tissue were obtained from rats after intraperitoneal injections with alcohol (75 mmol/kg body weight) with or without pretreatment with cyanamide (0.05 mmol/kg body weight), an aldehyde dehydrogenase inhibitor. Results: The amounts of MDA and unreduced AA adducts were found to be significantly increased in the heart of the rats treated with ethanol, cyanamide, or both, whereas no other adducts were detected in statistically significant quantities. Immunohistochemical studies for characterization of adduct distribution revealed sarcolemmal adducts of both MDA and AA in the rats treated with ethanol and cyanamide in addition to intracellular adducts, which were also present in the group treated with ethanol alone. Conclusions: These findings support the role of enhanced lipid peroxidation and the generation of protein-aldehyde condensates in vivo as a result of excessive ethanol intake. These findings may have implications in the molecular mechanisms of cardiac dysfunction in alcoholics.

Protein film voltammetry of Rhodobacter capsulatus xanthine dehydrogenase

Xanthine dehydrogenase (XDH) from the bacterium Rhodobacter capsulatus catalyzes the hydroxylation of xanthine to uric acid with NAD(+) as the electron acceptor. R. capsulatus XDH forms an (alphabeta)(2) heterotetramer and is highly homologous to homodimeric eukaryotic XDHs. The crystal structures of bovine XDH and R. capsulatus XDH showed that the two proteins have highly similar folds; however, R. capsulatus XDH is at least 5 times more active than bovine XDH and, unlike mammalian XDH, does not undergo the conversion to the oxidase form. Here we demonstrate electrocatalytic activity of the recombinant enzyme, expressed in Escherichia coli, while immobilized on an edge plane pyrolytic graphite working electrode. Furthermore, we have determined all redox potentials of the four cofactors (Mo-VI/V, Mo-V/IV, FAD/FADH, FADH/FADH(2) and two distinct [2Fe-2S](2+/+) clusters) using a combination of potentiometric and voltammetric methods. A novel feature identified in catalytic voltammetry of XDH concerns the potential for the onset of catalysis (ca. 400 mV), which is at least 600 mV more positive than that of the highest potential cofactor. This unusual observation is explained on the basis of a pterin-associated oxidative switch during voltammetry that precedes catalysis.

A protective effect of early pregnancy factor on experimental autoimmune encephalomyelitis induced in Lewis rats by inoculation with myelin basic protein

Experimental antoimmune encephalomyelitis (EAE) is an organ-specific autoimmune disease characterised by inflammation and demyelination of the central nervous system and is the best available animal model of multiple sclerosis (MS). Since previous studies have shown that EAE is less severe or is delayed in onset during pregnancy and that administration of the pregnancy hormone early pregnancy factor (EPF) down-regulates EAE, experiments in the present study were designed to explore further the role of EPF in EAE. By using the rosette inhibition test, the standard bioassay for EPF and, by semi-quantitative RT-PCR techniques, we have now shown that inflammatory cells from the spinal cord of rats with EAE can produce and secrete EPF, with production being greatest during recovery from disease. Administration of EPF to rats with EAE resulted in a significant increase in the expression of IL-4 and IL-10 mRNA and a significant decrease in IFN-gamma mRNA expression in spinal cord inflammatory cells. Encephalitogenic MBP-specific T cell lines were prepared from popliteal lymph nodes of rats with EAE. Proliferation assays using these cells demonstrated the ability of exogenous EPF to down-regulate the responses of T lymphocytes to MBP. (C) 2003 Elsevier B.V. All rights reserved.

Cultivo in vitro de Crambe abyssinica Hochst. : germinação, micropropagação, estabilidade genética e anatomia foliar

Crambe (Crambe abyssinica) pertence à família Brassicaceae, originário da Etiópia e principalmente destinado à produção de forragem (30 a 32% de proteína bruta). Atualmente, tem sido bastante cultivado visando à extração de óleo vegetal. Com os atuais incentivos à busca de fontes de energias renováveis, o cultivo de crambe vem ganhando papel de destaque na produção de biodiesel por suas diversas vantagens, como: (a) rápido ciclo de vida (colhida em torno de 90 dias); (b) alta produção de biomassa; (c) alta produtividade de sementes (1000 e 1500 kg ha-1); (d) menor custo de produção em relação a outras fontes oleaginosas, como, canola, girassol e soja; (e) um percentual de óleo total na semente entre 32 e 38%, superando, por exemplo, a soja; (f) potencial de fitorremediação, eficiente na descontaminação de arsênio, cromo e outros metais pesados; e (g) elevado percentual de ácido erúcico (50 a 60%) sendo útil na indústria de plástico e lubrificante. Devido aos poucos trabalhos realizados com crambe, abre-se um vasto campo de investigações científicas que tenham como objetivo desenvolver as potencialidades dessa cultura e, consequentemente, melhorar os aspectos agronômicos e tecnológicos para seu emprego na indústria de biodiesel. Nesse contexto, as técnicas de cultivo in vitro foram importantes tanto para a propagação massal, quanto como ferramenta para uma possível aplicação de outras técnicas biotecnológicas, contribuindo para uma produção homogênea, fiel e em larga escala. Portanto, este trabalho teve como objetivo geral avaliar as condições mais favoráveis à germinação, estabelecimento in vitro e micropropagação de Crambe abyssinica Hochst., além de verificar possíveis alterações genéticas e anatômicas, possibilitando a regeneração e produção de plântulas viáveis. Para a germinação e estabecimento in vitro de crambe, as condições mais favoráveis foram em meio B5 ou WPM, na presença ou ausência de pericarpo e na presença de luz. Na micropropagação dessa espécie, uma frequência satisfatória de regeneração de brotos foi obtida a partir de segmentos apicais utilizados como explante em meio contendo 5 μM de BAP (6- benzilaminopurina), e o alongamento foi satisfatório com 1 μM de GA3 (ácido giberélico). Os marcadores moleculares ISSR (Inter-Simples Sequence Repeats) utilizados para a análise da estabilidade genética indicaram que o segmento apical de crambe é um explante confiável para a micropropagação de plantas geneticamente verdadeiras (true-to-tipe), ou seja, mantém a estabilidade genética. As diversas fontes de citocininas e concentrações utilizadas neste trabalho não promoveram mudanças, no sentido de alterar a organização e/ou a espessura em relação ao controle, e as alterações observadas na estrutura e espessura das folhas dos tratamentos de aclimatização prejudicaram o processo de estabelecimento da plântula ex vitro. Contudo, existe a necessidade de um enraizamento e aclimatização eficiente para completa propagação in vitro de crambe. Portanto, este protocolo de regeneração de plantas in vitro de crambe pode ser útil no processo de criação e desenvolvimento de novas cultivares em um tempo mais curto e no melhoramento genético usando explantes apicais.

The influence of light spectra, UV-A, and growth regulators on the in vitro seed germination of Senecio cineraria DC.

This study was carried out to investigate the effects of light spectra, additional UV-A, and different growth regulators on the in vitro germination of Senecio cineraria DC. Seeds were surface-sterilized and inoculated in MS medium to evaluate the following light spectra: white, white plus UV-A, blue, green, red or darkness. The maximum germinability was obtained using MS0 medium under white light (30%) and MS + 0.3 mg L-1 GA3 in the absence of light (30.5%). S. cineraria seeds were indifferent to light. Blue and green lights inhibited germination. Different concentrations of gibberellic acid (GA3) (0.1; 0.4; 0.6; 0.8; 1.0 and 2.0 mg L-1) and indole-3-acetic acid IAA (0.1; 0.3 and 1.0 mg L-1) were evaluated under white light and darkness. No concentration of GA3 enhanced seed germination percentage under white light. However, when the seeds were maintained in darkness, GA3 improved germination responses in all tested concentrations, except at 1.0 mg L-1. Under white light, these concentrations also increased the germination time and reduced germination rate. Germination rate, under light or darkness, was lower using IAA compared with GA3.

An image processing application for quantification of protein aggregates in Caenorhabditis elegans

Protein aggregation became a widely accepted marker of many polyQ disorders, including Machado-Joseph disease (MJD), and is often used as readout for disease progression and development of therapeutic strategies. The lack of good platforms to rapidly quantify protein aggregates in a wide range of disease animal models prompted us to generate a novel image processing application that automatically identifies and quantifies the aggregates in a standardized and operator-independent manner. We propose here a novel image processing tool to quantify the protein aggregates in a Caenorhabditis elegans (C. elegans) model of MJD. Confocal mi-croscopy images were obtained from animals of different genetic conditions. The image processing application was developed using MeVisLab as a platform to pro-cess, analyse and visualize the images obtained from those animals. All segmenta-tion algorithms were based on intensity pixel levels.The quantification of area or numbers of aggregates per total body area, as well as the number of aggregates per animal were shown to be reliable and reproducible measures of protein aggrega-tion in C. elegans. The results obtained were consistent with the levels of aggrega-tion observed in the images. In conclusion, this novel imaging processing applica-tion allows the non-biased, reliable and high throughput quantification of protein aggregates in a C. elegans model of MJD, which may contribute to a significant improvement on the prognosis of treatment effectiveness for this group of disor-ders

Automatic segmentation and 3D feature extraction of protein aggregates in Caenorhabditis Elegans

In the last years, it has become increasingly clear that neurodegenerative diseases involve protein aggregation, a process often used as disease progression readout and to develop therapeutic strategies. This work presents an image processing tool to automatic segment, classify and quantify these aggregates and the whole 3D body of the nematode Caenorhabditis Elegans. A total of 150 data set images, containing different slices, were captured with a confocal microscope from animals of distinct genetic conditions. Because of the animals’ transparency, most of the slices pixels appeared dark, hampering their body volume direct reconstruction. Therefore, for each data set, all slices were stacked in one single 2D image in order to determine a volume approximation. The gradient of this image was input to an anisotropic diffusion algorithm that uses the Tukey’s biweight as edge-stopping function. The image histogram median of this outcome was used to dynamically determine a thresholding level, which allows the determination of a smoothed exterior contour of the worm and the medial axis of the worm body from thinning its skeleton. Based on this exterior contour diameter and the medial animal axis, random 3D points were then calculated to produce a volume mesh approximation. The protein aggregations were subsequently segmented based on an iso-value and blended with the resulting volume mesh. The results obtained were consistent with qualitative observations in literature, allowing non-biased, reliable and high throughput protein aggregates quantification. This may lead to a significant improvement on neurodegenerative diseases treatment planning and interventions prevention

Production of basic potato seed minitubers in substrate and different nitrogen rates

The optimal dose of nitrogen (N) in potato crop depends on the production system. The objective of this study was to determine the optimal dose of N for the production of basic potato seed minitubers and evaluate the effect of N rates on physiological and nitrogen indices in the youngest fully developed leaf (fourth leaf) and in the oldest leaf of the plants at 60 days after planting. The experiment was conducted in a greenhouse at the Departamento de Fitotecnia da Universidade Federal de Viçosa. The treatments consisted of five N rates (0, 45, 90, 180 and 360 mg dm-3), with 10% of each dose applied at planting and the remainder through irrigation water, daily, for 30 days. The nitrogen rates positively influenced the physiological indices (length, width, leaf area, number of leaves, fresh mass and dry mass) and nitrogen (level and content of N and N-NO³ in the dry mass and SPAD) both in the fourth leaf and in the oldest leaf. Likewise, the N rates positively influenced the number and mass of harvested tubers. The largest number (5.44 tubers/plant) and the maximum mass of tubers (243.5 g/plant) were obtained with 360.0 and 332.9 mg N dm-3, respectively. Therefore, the mass and number of tubers were not optimized by the same N rate. The critical SPAD index was 38.8 in the fourth leaf, which was more sensitive to the effect of N rates than the oldest leaf.

Chilling requirement for seed germination and phenological observations on peach cultivars

In subtropical climate areas, the models and methods proposed to evaluate the chilling requirement of temperate fruit crops often do not provide satisfactory results, thus calling for the development of alternative techniques. The aim of this study was to evaluate the correlations between some phonological traits and chilling requirement for seed germination of 18 peach cultivars and one nectarine cultivar. Two experiments were installed separately for the correlation studies. In experiment 1, the phenological traits were observed in the field, while in experiment 2, the chilling requirement for 50 and 100% seed germination of each cultivar was assessed. The number of days for beginning of bloom (r = 0.70**, 0.61**) and full bloom (r = 0.72**, 0.76**) were both significantly correlated with the number of chilling units for 50% and 100% germination of seeds. The number of days for beginning of budding and dormancy break were both significantly correlated with the number of chilling units for 50% and 100% germination (r = 0.48*, 0.50*, respectively). However, the same significant effect for these phenological traits was not found between chilling units and 50% germination of seeds, as well as between chilling units and harvest dates.

Lignification of the plant and seed quality of RR soybeans sprayed with herbicide glyphosate

Differences in levels of lignin in the plant between conventional and transgenic cultivars RR has been reported by several authors, however, there are few studies evaluating the influence of spraying of glyphosate on the lignin in the plant and RR soybean seeds. The aim of this study was to evaluate the physiological quality of RR transgenic soybean seeds and the lignin contents of plants sprayed with the herbicide glyphosate. The assays were conducted both in greenhouse and field in the municipality of Lavras, MG, in the agricultural year 2007/08. The experiment was arranged in a splitplot design with four replicates, considering the treatments hand weeding and herbicide glyphosate as plots, and five RR soybean cultivars (BRS 245 RR, BRS 247 RR, Valiosa RR, Silvânia RR and Baliza RR) as splitplots. In the greenhouse, the cultivars tested were BRS 245 RR and Valiosa RR in a randomized block design with four replicates. The sprayings were carried out at stages V3, V7 and early R5 (3L/ha). The 1000 seed weight, mechanical injury, germination and germination velocity index, emergence velocity index, accelerated aging, electrical conductivity and water soaking seed test, lignin content in the seed coat, in the stem and legumes were determined. The spraying of glyphosate herbicide, in greenhouse and field, did not alter the physiological quality of seeds and the lignin contents in the plant.

A study on the encryption algorithm for RFID tag (SEED : 8 Rounds × 64 bits block)

Although it is always weak between RFID Tag and Terminal in focus of the security, there are no security skills in RFID Tag. Recently there are a lot of studying in order to protect it, but because it has some physical limitation of RFID, that is it should be low electric power and high speed, it is impossible to protect with the skills. At present, the methods of RFID security are using a security server, a security policy and security. One of them the most famous skill is the security module, then they has an authentication skill and an encryption skill. In this paper, we designed and implemented after modification original SEED into 8 Round and 64 bits for Tag.

TBCCD1, a new centrosomal protein, is required for centrosome and Golgi apparatus positioning

In animal cells the centrosome is positioned at the cell centre in close association with the nucleus. The mechanisms responsible for this are not completely understood. Here, we report the first characterization of human TBCC-domain containing 1 (TBCCD1), a protein related to tubulin cofactor C. TBCCD1 localizes at the centrosome and at the spindle midzone, midbody and basal bodies of primary and motile cilia. Knockdown of TBCCD1 in RPE-1 cells caused the dissociation of the centrosome from the nucleus and disorganization of the Golgi apparatus. TBCCD1-depleted cells are larger, less efficient in primary cilia assembly and their migration is slower in wound-healing assays. However, the major microtubule-nucleating activity of the centrosome is not affected by TBCCD1 silencing. We propose that TBCCD1 is a key regulator of centrosome positioning and consequently of internal cell organization.