Overall Picture Of Expressed Heat Shock Factors In Glycine Max, Lotus Japonicusand Medicago Truncatula

Heat shock (HS) leads to the activation of molecular mechanisms, known as HS-response, that prevent damage and enhance survival under stress. Plants have a flexible and specialized network of Heat Shock Factors (HSFs), which are transcription factors that induce the expression of heat shock proteins. The present work aimed to identify and characterize the Glycine maxHSF repertory in the Soybean Genome Project (GENOSOJA platform), comparing them with other legumes (Medicago truncatulaand Lotus japonicus) in view of current knowledge of Arabidopsis thaliana. The HSF characterization in leguminous plants led to the identification of 25, 19 and 21 candidate ESTs in soybean, Lotusand Medicago, respectively. A search in the SuperSAGE libraries revealed 68 tags distributed in seven HSF gene types. From the total number of obtained tags, more than 70% were related to root tissues (water deficit stress libraries vs.controls), indicating their role in abiotic stress responses, since the root is the first tissue to sense and respond to abiotic stress. Moreover, as heat stress is related to the pressure of dryness, a higher HSF expression was expected at the water deficit libraries. On the other hand, expressive HSF candidates were obtained from the library inoculated with Asian Soybean Rust, inferring crosstalk among genes associated with abiotic and biotic stresses. Evolutionary relationships among sequences were consistent with different HSF classes and subclasses. Expression profiling indicated that regulation of specific genes is associated with the stage of plant development and also with stimuli from other abiotic stresses pointing to the maintenance of HSF expression at a basal level in soybean, favoring its activation under heat-stress conditions. © 2012, Sociedade Brasileira de Genética.35SUPPL.1247259Altschul, S.F., Gish, W., Miller, W., Myers, E.W., Lipman, D.J., Basic local alignment search tool (1990) J Mol Biol, 215, pp. 403-410Baniwal, S.K., Chan, K.Y., Scharf, K.-D., Nover, L., Role of heat stress transcription factor HsfA5 as specific repressor of HsfA4* (2007) J Biol Chem, 282, pp. 3605-3613Bharti, K., Schimidt, E., Lyck, R., Bublak, D., Scharf, K.-D., Isolation and characterization of HsfA3, a new heat stress transcription factor of Lycopersicon peruvianum (2000) Plant J, 22, pp. 355-365Bharti, K., von Koskull-Döring, P., Bharti, S., Kumar, P., Tintschl-Körbitzer, A., Treuter, E., Nover, L., Tomato heat stress transcription factor HsfB1 represents a novel type of general transcription coactivator with a histone-like motif interacting with HAC1/CBP (2004) Plant Cell, 16, pp. 1521-1535Efeoglu, B., Heat shock proteins and heat shock response in plants (2009) G U J Sci, 22, pp. 67-75Eisen, M.B., Spellman, P.T., Brown, P.O., Botstein, D., Cluster analysis and display of genome-wide expression patterns (1998) Proc Natl Acad Sci USA, 95, pp. 14863-14868Fehr, W.R., Caviness, C.E., Burmood, D.T., Pennington, I.S., Stage of development descriptions for soybeans, Glycine max (L.) Merrill (1971) Crop Sci, 11, pp. 929-931Fehr, W.R., Caviness, C.E., (1977) Stage of Soybean Development, p. 12. , Special Report n. 80. Ames, Iowa State University of Science and Technology, IowaGlombitza, S., Dubuis, P.-H., Thulke, O., Welzl, G., Bovet, L., Götz, M., Affenzeller, M., Asnaghi, C., Crosstalk and differential response to abiotic and biotic stressors reflected at the transcriptional level of effector genes from secondary metabolism (2004) Plant Mol Biol, 54, pp. 817-835Heerklotz, D., Doring, P., Bonzelius, F., Winkelhaus, S., Nover, L., The balance of nuclear import and export determines the intracellular distribution and function of tomato heat stress transcription factor HsfA2 (2001) Mol Cell Biol, 21, pp. 1759-1768Hoagland, D., Arnon, D.I., The water culture method for growing plants without soil (1950) Calif Agric Exp Stn Circ, 347, pp. 1-32Hsu, S.-F., Lai, H.-C., Jinn, T.-L., Cytosol-localized heat shock factor-binding protein, AtHSBP, functions as a negative regulator of heat shock response by translocation to the nucleus and is required for seed development in Arabidopsis (2010) Plant Physiol, 153, pp. 773-784Hu, W., Hu, G., Han, B., Genome-wide survey and expression profiling of heat shock proteins and heat shock factors revealed overlapped and stress specific response under abiotic stresses in rice (2009) Plant Sci, 176, pp. 583-590Kido, E.A., Barbosa, P.K., Ferreira Neto, J.C.R., Pandolfi, V., Houllou-Kido, L.M., Crovella, S., Benko-Iseppon, A.M., Identification of plant protein kinases in response to abiotic and biotic stresses using SuperSAGE (2011) Curr Prot Pept Sci, 12, pp. 643-656Kotak, S., Port, M., Ganguli, A., Bicker, F., von Koskull-Doring, P., Characterization of C-terminal domains of Arabidopsis heat stress transcription factors (Hsfs) and identification of a new signature combination of plant class a Hsfs with AHA and NES motifs essential for activator function and intracellular localization (2004) Plant J, 39, pp. 98-112Kotak, S., Larkindale, J., Lee, U., von Koskull-Doring, P., Vierling, E., Scharf, K.D., Complexity of the heat stress response in plants (2007) Curr Opin Plant Biol, 10, pp. 310-316Li, H.-Y., Chang, C.-S., Lu, L.-S., Liu, C.-A., Chan, M.-T., Charng, Y.-Y., Over-expression of Arabidopsis thaliana heat shock factor gene (AtHsfA1b) enhances chilling tolerance in transgenic tomato (2004) Bot Bull Acad Sin, 44, pp. 129-140Li, M., Berendzen, K.W., Schoffl, F., Promoter specificity and interactions between early and late Arabidopsis heat shock factors (2010) Plant Mol Biol, 73, pp. 559-567McClean, P.E., Mamidi, S., McConnell, M., Chikara, S., Lee, R., Synteny mapping between common bean and soybean reveals extensive blocks of shared loci (2010) BMC Genomics, 11, pp. e184Miller, G., Mittler, R., Could heat shock transcription factors function as hydrogen peroxide sensors in plant? (2006) Ann Bot, 98, pp. 279-288Mittal, D., Chakrabarti, S., Sarkar, A., Singh, A., Grover, A., Heat shock factor gene family in rice: Genomic organization and transcript expression profiling in response to high temperature, low temperature and oxidative stresses (2009) Plant Physiol Biochem, 47, pp. 785-795Mochida, K., Yoshida, T., Sakurai, T., Yamaguchi-Shinozaki, K., Shinozaki, K., Tran, L.-S.P., In silico analysis of transcription factor repertoire and prediction of stress responsive transcription factors in soybean (2009) DNA Res, 16, pp. 353-369Mochida, K., Yoshida, T., Sakurai, T., Yamaguchi-Shinozaki, K., Shinozaki, K., Tran, L.-S.P., LegumeTFDB: An in-tegrative database of Glycine max, Lotus japonicus and Medicago truncatula transcription factors (2009) Bioinformatics, 26, pp. 290-291Nascimento, L.C., Costa, G.G.L., Binneck, E., Pereira, G.A.G., Caraz-Zolle, M.F., A web-based bioinformatics interface applied to Genosoja Project: Databases and pipelines (2012) Genet Mol Biol, 35 (SUPPL. 1), pp. 203-211Nover, L., Bharti, K., Doring, P., Mishra, S.K., Ganguli, A., Scharf, K.-D., Arabidopsis and the heat stress transcription factor world: How many heat stress transcription factors do we need? (2001) Cell Stress Chap, 6, pp. 177-189Pirkkala, L., Nykanen, I., Sistonen, L., Roles of the heat shock transcription factors in regulation of the heat shock response and beyond (2001) FASEB J, 15, pp. 1118-1131Ruelland, E., Zachowski, A., How plants sense temperature (2010) Environ Exp Bot, 69, pp. 225-232Sato, Y., Yokoya, S., Enhanced tolerance to drought stress in transgenic rice plants overexpressing a small heat-shock protein, sHSP17.7 (2008) Plant Cell Rep, 27, pp. 329-334Scharf, K.-D., Rose, S., Thierfelder, J., Nover, L., Two cDNAs for tomato heat stress transcription factors (1993) Plant Physiol, 102, pp. 1355-1356Scharf, K.-D., Rose, S., Zott, W., Schoffl, F., Nover, L., Three tomato genes code for heat stress transcription factors with a regionofremarkable homology to the DNA-binding domain of the yeast HSF (1990) EMBO J, 9, pp. 4495-4501Schöff, F., Prändl, R., Reindl, A., Regulation of the heat-shock response (1998) Plant Physiol, 117, pp. 1135-1141Sung, D.-Y., Kaplan, F., Lee, K.-J., Guy, C.L., Acquired tolerance to temperature extremes (2003) Trends Plant Sci, 8, pp. 179-187Swindell, W.R., Huebner, M., Weber, A.P., Transcriptional profiling of Arabidopsis heat shock proteins and transcription factors reveals extensive overlap between heat and non-heat stress response pathways (2007) BMC Genomics, 8, pp. e125Tamura, K., Peterson, D., Peterson, N., Stecher, G., Nei, M., Kumar, S., MEGA5: Molecular Evolutionary Genetics Analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods (2011) Mol Biol Evol, 28, pp. 2731-2739Treshow, M., (1970) Environment and Plant Response, p. 421. , McGraw-Hill Company, New YorkTreuter, E., Nover, L., Ohme, K., Scharf, K.-D., Promoter specificity and deletion analysis of three tomato heat stress transcription factors (1993) Mol Gen Genet, 240, pp. 113-125Yamada, K., Fukao, Y., Hayashi, M., Fukazawa, M., Suzuki, I., Nishimura, M., Cytosolic HSP90 regulated the heat shock response that is responsible for heat acclimation in Arabidopsis thaliana (2007) J Biol Chem, 282, pp. 37794-3780

Antiproliferative effects of lectins from Canavalia ensiformis and Canavalia brasiliensis in human leukemia cell lines

AbstractThe antiproliferative activity of lectins Canavalia ensiformis (ConA) and Canavalia brasiliensis (ConBr) were studied using human leukemia MOLT-4 and HL-60 cell lines. It was revealed that both ConA and ConBr were markedly cytotoxic to cells using MTT and NAC assays. The IC50 values were approximately 3 and 20μg/mL for ConA and ConBr, respectively, for both MOLT-4 and HL-60 cells. However, in normal human peripheral blood lymphocytes, the lectins were not cytotoxic, even when tested at concentrations as high as 200μg/ml. Using comet assay, the lectins produced a rate of DNA damage exceeding 80% in MOLT-4 and HL-60 cells. Fluorescence analysis revealed the morphology characteristic of apoptosis, with low concentrations of apoptotic bodies and fragmented DNA (5μg/ml). Flow cytometric analysis demonstrated an accumulation of cells in the sub-G1 cell cycle that is characteristic of DNA fragmentation, and a decrease in membrane integrity at high concentrations. Lastly, we evaluated the alterations in mitochondrial potential that reduced after treatment with lectins. Our results indicate that ConA and ConBr inhibited cell proliferation selectively in tumor cells and that apoptosis was the main death mechanism. Therefore, lectins can be considered a class of molecules with a high antitumor activity potential

An Overall Evaluation Of The Resistance (r) And Pathogenesis-related (pr) Super Families In Soybean, As Compared With Medicago And Arabidopsis

Plants have the ability to recognize and respond to a multitude of pathogens, resulting in a massive reprogramming of the plant to activate defense responses including Resistance (R) and Pathogenesis-Related (PR) genes. Abiotic stresses can also activate PR genes and enhance pathogen resistance, representing valuable genes for breeding purposes. The present work offers an overview of soybean Rand PR genes present in the GENOSOJA (Brazilian Soybean Genome Consortium) platform, regarding their structure, abundance, evolution and role in the plant-pathogen metabolic pathway, as compared with Medicago and Arabidopsis. Searches revealed 3,065 R candidates (756 in Soybean, 1,142 in Medicago and 1,167 in Arabidopsis), and PR candidates matching to 1,261 sequences (310, 585 and 366 for the three species, respectively). The identified transcripts were also evaluated regarding their expression pattern in 65 libraries, showing prevalence in seeds and developing tissues. Upon consulting the Super SAGE libraries, 1,072 Rand 481 PR tags were identified in association with the different libraries. Multiple alignments were generated forXa21andPR-2genes, allowing inferences about their evolution. The results revealed interesting insights regarding the variability and complexity of defense genes in soybean, as compared with Medicago and Arabidopsis. © 2012, Sociedade Brasileira de Genética.35SUPPL.1260271Alberts, B., Johnson, A., Lewis, J., Raff, M., Roberts, K., Walter, P., (2002) Molecular Biology of the Cell, p. 1616. , 4th edition. 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'avenae' (1994) Theor Appl Genet, 89, pp. 831-837Salmeron, J.M., Oldroyd, G.E.D., Romens, C.M.T., Scofield, S.R., Kim, H.S., Lavelle, D.T., Dahlbeck, D., Staskawicz, B.J., Tomato Prf is a member of the leucine rich repeats class of plant disease resistance genes and lies embedded within the Pto kinase gene cluster (1996) Cell, 86, pp. 123-133Shoemaker, R.C., Schlueter, J., Doyle, J.J., Paleopolyploidy and gene duplication in soybean and other legumes (2006) Curr Opin Plant Biol, 9, pp. 104-109Song, W.Y., Pi, L.Y., Wang, G.L., Gardner, J., Holsten, T., Ronald, P.C., Evolution of the rice Xa21 disease resistance genes family (1997) Plant Cell, 9, pp. 1279-1287Song, W.Y., Wang, G.L., Kim, H.S., Pi, L.Y., Gardner, J., Wang, B., Holsten, T., Fauquet, C., A receptor kinase-like protein encoded by the rice disease resistance gene Xa21 (1995) Science, 270, pp. 1804-1806Sparla, F., Rotino, L., Valgimigli, M.C., Pupillo, P., Trost, P., Systemic resistance induced by benzothisdizole in pear inoculated with the agent of fire blight (2004) Sci Hortic, 101, pp. 269-279Tamura, K., Dudley, J., Nei, M., Kumar, S., MEGA4: Molecular Evolutionary Genetics Analysis (MEGA) software ver. 4.0 (2007) Mol Biol Evol, 24, pp. 1596-1599Tang, X., Xie, M., Kim, Y.J., Zhou, J., Klessing, D.F., Martin, G.B., Overexpression of Pto activates defense responses and confers broad resistance (1999) Plant Cell, 11, pp. 15-29Thiel, T., Graner, A., Waugh, R., Grosse, I., Close, T.J., Stein, N., Evidence and evolutionary of ancient whole-genome duplication in barley predating the divergence from rice (2009) BMC Evol Biol, 9, pp. 209-227Tornero, P., Gadea, J., Conejero, V., Vera, P., Two PR-1 genes from tomato are differentially regulated and reveal a novel mode of expression for a pathogenesis-related gene during the hypersensitive response and development (1997) Plant Microbe Interact, 10, pp. 624-634Van-Loon, L.C., Geraats, B.P.J., Linthorst, H.J.M., Ethylene as a modulator of disease resistance in plants (2006) Trends Plant Sci, 11, pp. 184-191Van-Loon, L.C., Pierpoint, W.S., Boller, T., Conejero, V., Recommendations for naming plant pathogenesis-related proteins (1999) Plant Mol Biol Rep, 12, pp. 245-264Velazhahan, R., Muthukrishnan, S., Transgenic tobacco plants constitutively overexpressing a rice thaumatin-like protein (PR-5) show enhanced resistance to Alternaria alternata (2003) Plant Biol, 47, pp. 347-354Vergne, E., Grand, X., Ballini, R., Chalvon, V., Saindrenan, P., Tharreau, D., Nottéghem, J.-L., Morel, J.-B., Preformed expression of defense is a hallmark of partial resistance to rice blast fungal pathogen Magnaporthe oryzae (2010) BMC Plant Biol, 10, pp. e206Wanderley-Nogueira, A.C., Mota, N., Lima-Morais, D., Silva, L.C.B., Silva, A.B., Benko-Iseppon, A.M., Abundance and diversity of resistance (R) genes in the sugarcane trans-criptome (2007) Genet Mol Res, 6, pp. 866-889Wang, G.L., Holsten, T.E., Song, W.Y., Wang, H.P., Ronald, P.C., Construction of a rice bacterial artificial chromosome library and identification of clones linked to the Xa21 disease resistance locus (1995) Plant J, 7, pp. 525-533Wendell, J., Genome evolution in polyploids (2000) Plant Mol Biol, 42, pp. 225-249Weng, J.K., Banks, J.A., Chapple, C., Parallels in lignin biosynthesis: A study in Selaginella moellendorffii reveals convergence across 400 million years of evolution (2008) Comm Int Biol, 1, pp. 20-22Wilkstrom, N., Savolainen, V., Chase, M.W., Evolution of the angiosperms: Calibrating the family tree (2001) Proc Soc Biol Sci, 268, pp. 2211-2220Zeier, J., Pink, B., Mueller, M.J., Berger, S., Light conditions influence specific defense responses in incompatible plant-pathogen interactions: Uncoupling systemic resistance from salicylic acid and PR-1 accumulation (2004) Planta, 219, pp. 673-68

Characteristic Time and Maximum Mixedness: Single Mode Gaussian States in Dissipative Channels

We derive an upper limit for the mixedness of single bosonic mode gaussian states propagating in dissipative channels. It is a function of the initial squeezing and temperature of the channel only. Moreover the time at which von Neumann's entropy reaches its maximum value coincides with that of complete loss of coherence, thus defining a quantum-classical transition.Comment: Accepted versio

Experimental implementation of a NMR entanglement witness

Entanglement witnesses (EW) allow the detection of entanglement in a quantum system, from the measurement of some few observables. They do not require the complete determination of the quantum state, which is regarded as a main advantage. On this paper it is experimentally analyzed an entanglement witness recently proposed in the context of Nuclear Magnetic Resonance (NMR) experiments to test it in some Bell-diagonal states. We also propose some optimal entanglement witness for Bell-diagonal states. The efficiency of the two types of EW's are compared to a measure of entanglement with tomographic cost, the generalized robustness of entanglement. It is used a GRAPE algorithm to produce an entangled state which is out of the detection region of the EW for Bell-diagonal states. Upon relaxation, the results show that there is a region in which both EW fails, whereas the generalized robustness still shows entanglement, but with the entanglement witness proposed here with a better performance

Evaluation of MTHFR C677T gene polymorphism and homocysteine level in coronary atherosclerotic disease

OBJECTIVE: The aim of this study is to determine the prevalence of C677T methylenetetrahydrofolate reductase (MTHFR) polymorphism and correlate it with plasma homocysteine levels in coronary artery disease (CAD). METHODS: Ninety-three patients with documented CAD from Hospital Universitário Oswaldo Cruz (Recife, PE, Brazil) and 108 healthy controls were evaluated. Homocysteine and folate levels were determined by HPLC and chemoluminescence, respectively, and lipid profile was considered. Genotyping was done by RFLP/PCR. RESULTS: The groups were homogeneous for the C677T polymorphisms. The homocysteine level in cases (11.7 µmol/L) was statistically different from that observed in controls (8.84 µmol/L, p< 0.05). It was also observed that 72% of the patients had homocysteine values above12 µmol/L while the control group presented only 32% in this range. There was no relationship between homozigosity for the C677T polymorphism and the homocysteine level (p= 0.634). We noticed statistical differences between folate levels from patients and controls (6.22 and 7.69 ng/dL, p< 0.05, respectively). However, there was no correlation between homocysteine and folate concentrations in the entire group (r= -0.202). Comparing cases and controls, the odds ratio (OR) when homocysteine is high and folate is low was OR= 11.9; CI 95%= 4.16-34.42, p< 0.01. CONCLUSION: A lack of correlation between C677T mutation and homocysteine level suggests that environmental factors and others genetic factors seem to exert more influence on homocysteine level in this population.OBJETIVO: O objetivo deste trabalho é determinar a prevalência do polimorfismo C677T do gene metilenotetraidrofolato redutase (MTHFR) e associá-la com a concentração plasmática de homocisteína plasmática na doença arterial coronariana (DAC). MÉTODOS: Foram avaliados 93 pacientes com DAC documentada, atendidos no Hospital Universitário Oswaldo Cruz (Recife, PE, Brasil), e 108 controles sem a doença. Foram determinados os perfis lipídicos de pacientes e controles. As concentrações plasmáticas de homocisteína e folato foram determinadas por HPLC e quimioluminescência, respectivamente. A genotipagem foi realizada por RFLP/PCR. RESULTADOS: Os grupos de pacientes e controles foram homogêneos quanto aos perfis genéticos do polimorfismo investigado. Nos pacientes, as concentrações plasmáticas médias de homocisteina (11,7 ± 4,4 µmol/L) e de folato (6,22 ± 3,0 ng/dL) foram estatisticamente diferentes daquelas observadas nos controles (8,84 ± 3,2 µmol/L e 7,69 ± 3,1 ng/dL, respectivamente), ao nível de significância de 0,05. Entretanto, não houve correlação entre concentração plasmática de homocisteína e folato nos pacientes (r= -0,202). Não foi observada associação entre a homozigosidade 677TT para MTHFR e a concentração plasmática de homocisteína sérica (p= 0,634). A comparação dos casos e controles que apresentaram simultaneamente alta concentração plasmática de homocisteína e baixa concentração de folato, resultou numa razão de chance superior à de cada variável analisada independentemente (RC= 11,9; IC 95%= 4,16-34,42, p< 0,01). CONCLUSÕES: A mutação C677T não parece ser um fator genético importante capaz de explicar a hiperhomocisteinemia moderada observada nos pacientes com DAC. Outros fatores, ambientais e genéticos, devem ser investigados.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Universidade Federal de São Paulo (UNIFESP) Departamento de Pediatria Laboratório de Erros Inatos de MetabolismoUniversidade de Pernambuco ICB Departamento de Ciências FisiológicasUniversidade de São Paulo Faculdade de Medicina de Ribeirão Preto Hospital das ClínicasUNIFESP, Depto. de Pediatria Laboratório de Erros Inatos de MetabolismoSciEL

Cultura e desenvolvimento humano sustentável

A cultura é cada vez mais o centro das políticas locais, pela importância que vem revelando no contexto de um paradigma de desenvolvimento humano integral. Para além da atenção que lhe é dirigida graças à sua intima conexão a fatores de índole económica, social e urbana; a sua relevância deve-se essencialmente às relações intrínsecas que mantém com as questões da identidade, da memória, da criatividade, da ciência e do pensamento e conhecimento crítico. Optámos por dividir este livro em três capítulos de modo a permitir uma sequência de leitura, desde as temáticas gerais de grande escala – da União Europeia e da Globalização, até ao cerne do problema que em nosso entendimento urge solucionar nas cidades: a efetiva e definitiva integração da dimensão cultural nas estratégias territoriais de desenvolvimento humano sustentável.N/