The gene for albicidin detoxification from Pantoea dispersa encodes an esterase and attenuates pathogenicity of Xanthomonas albilineans to sugarcane

Albicidin phytotoxins are pathogenicity factors in a devastating disease of sugarcane known as leaf scald, caused by Xanthomonas albilineans. A gene (albD) from Pantoea dispersa has been cloned and sequenced and been shown to code for a peptide of 235 amino acids that detoxifies albicidin. The gene shows no significant homology at the DNA or protein level to any known sequence, but the gene product contains a GxSxG motif that is conserved in serine hydrolases. The AlbD protein, purified to homogeneity by means of a glutathione S-transferase gene fusion system, showed strong esterase activity on p-nitrophenyl butyrate and released hydrophilic products during detoxification of albicidins. AlbD hydrolysis of p-nitrophenyl butyrate and detoxification of albicidins required no complex cofactors. Both processes were strongly inhibited by phenylmethylsulfonyl fluoride, a serine enzyme inhibitor. These data strongly suggest that AlbD is an albicidin hydrolase. The enzyme detoxifies albicidins efficiently over a pH range from 5.8 to 8.0, with a broad temperature optimum from 15 to 35°C. Expression of albD in transformed X. albilineans strains abolished the capacity to release albicidin toxins and to incite disease symptoms in sugarcane. The gene is a promising candidate for transfer into sugarcane to confer a form of disease resistance.

Rubisco activase constrains the photosynthetic potential of leaves at high temperature and CO2

Net photosynthesis (Pn) is inhibited by moderate heat stress. To elucidate the mechanism of inhibition, we examined the effects of temperature on gas exchange and ribulose 1,5-bisphosphate carboxylase/oxygenase (Rubisco) activation in cotton and tobacco leaves and compared the responses to those of the isolated enzymes. Depending on the CO2 concentration, Pn decreased when temperatures exceeded 35–40°C. This response was inconsistent with the response predicted from the properties of fully activated Rubisco. Rubisco deactivated in leaves when temperature was increased and also in response to high CO2 or low O2. The decrease in Rubisco activation occurred when leaf temperatures exceeded 35°C, whereas the activities of isolated activase and Rubisco were highest at 42°C and >50°C, respectively. In the absence of activase, isolated Rubisco deactivated under catalytic conditions and the rate of deactivation increased with temperature but not with CO2. The ability of activase to maintain or promote Rubisco activation in vitro also decreased with temperature but was not affected by CO2. Increasing the activase/Rubisco ratio reduced Rubisco deactivation at higher temperatures. The results indicate that, as temperature increases, the rate of Rubisco deactivation exceeds the capacity of activase to promote activation. The decrease in Rubisco activation that occurred in leaves at high CO2 was not caused by a faster rate of deactivation, but by reduced activase activity possibly in response to unfavorable ATP/ADP ratios. When adjustments were made for changes in activation state, the kinetic properties of Rubisco predicted the response of Pn at high temperature and CO2.

Hyperphosphorylation induces self-assembly of τ into tangles of paired helical filaments/straight filaments

The microtubule-associated protein τ is a family of six isoforms that becomes abnormally hyperphosphorylated and accumulates in the form of paired helical filaments (PHF) in the brains of patients with Alzheimer's disease (AD) and patients with several other tauopathies. Here, we show that the abnormally hyperphosphorylated τ from AD brain cytosol (AD P-τ) self-aggregates into PHF-like structures on incubation at pH 6.9 under reducing conditions at 35°C during 90 min. In vitro dephosphorylation, but not deglycosylation, of AD P-τ inhibits its self-association into PHF. Furthermore, hyperphosphorylation induces self-assembly of each of the six τ isoforms into tangles of PHF and straight filaments, and the microtubule binding domains/repeats region in the absence of the rest of the molecule can also self-assemble into PHF. Thus, it appears that τ self-assembles by association of the microtubule binding domains/repeats and that the abnormal hyperphosphorylation promotes the self-assembly of τ into tangles of PHF and straight filaments by neutralizing the inhibitory basic charges of the flanking regions.

Potentiation of capsaicin receptor activity by metabotropic ATP receptors as a possible mechanism for ATP-evoked pain and hyperalgesia

The capsaicin (vanilloid) receptor, VR1, is a sensory neuron-specific ion channel that serves as a polymodal detector of pain-producing chemical and physical stimuli. It has been proposed that ATP, released from different cell types, initiates the sensation of pain by acting predominantly on nociceptive ionotropic purinoceptors located on sensory nerve terminals. In this study, we examined the effects of extracellular ATP on VR1. In cells expressing VR1, ATP increased the currents evoked by capsaicin or protons through activation of metabotropic P2Y1 receptors in a protein kinase C-dependent pathway. The involvement of Gq/11-coupled metabotropic receptors in the potentiation of VR1 response was confirmed in cells expressing both VR1 and M1 muscarinic acetylcholine receptors. In the presence of ATP, the temperature threshold for VR1 activation was reduced from 42°C to 35°C, such that normally nonpainful thermal stimuli (i.e., normal body temperature) were capable of activating VR1. This represents a novel mechanism through which the large amounts of ATP released from damaged cells in response to tissue trauma might trigger the sensation of pain.

Nuclear-Gene Mutations Suppress a Defect in the Expression of the Chloroplast-Encoded Large Subunit of Ribulose-1,5-Bisphosphate Carboxylase/Oxygenase1

The green alga Chlamydomonas reinhardtii mutant 76–5EN lacks photosynthesis because of a nuclear-gene mutation that specifically inhibits expression of the chloroplast gene encoding the large subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco; EC 4.1.1.39). Photosynthesis-competent revertants were selected from mutant 76–5EN to explore the possibility of increasing Rubisco expression. Genetic analysis of 10 revertants revealed that most arose from suppressor mutations in nuclear genes distinct from the original 76–5EN mutant gene. The revertant strains have regained various levels of Rubisco holoenzyme, but none of the suppressor mutations increased Rubisco expression above the wild-type level in either the presence or absence of the 76–5EN mutation. One suppressor mutation, S107–4B, caused a temperature-conditional, photosynthesis-deficient phenotype in the absence of the original 76–5EN mutation. The S107–4B strain was unable to grow photosynthetically at 35°C, but it expressed a substantial level of Rubisco holoenzyme. Whereas the 76–5EN gene encodes a nuclear factor that appears to be required for the transcription of the Rubisco large-subunit gene, the S107–4B nuclear gene may be required for the expression of other chloroplast genes.

Ras Regulates the Polarity of the Yeast Actin Cytoskeleton through the Stress Response Pathway

Polarized growth in yeast requires cooperation between the polarized actin cytoskeleton and delivery of post-Golgi secretory vesicles. We have previously reported that loss of the major tropomyosin isoform, Tpm1p, results in cells sensitive to perturbations in cell polarity. To identify components that bridge these processes, we sought mutations with both a conditional defect in secretion and a partial defect in polarity. Thus, we set up a genetic screen for mutations that conferred a conditional growth defect, showed synthetic lethality with tpm1Δ, and simultaneously became denser at the restrictive temperature, a hallmark of secretion-defective cells. Of the 10 complementation groups recovered, the group with the largest number of independent isolates was functionally null alleles of RAS2. Consistent with this, ras2Δ and tpm1Δ are synthetically lethal at 35°C. We show that ras2Δ confers temperature-sensitive growth and temperature-dependent depolarization of the actin cytoskeleton. Furthermore, we show that at elevated temperatures ras2Δ cells are partially defective in endocytosis and show a delocalization of two key polarity markers, Myo2p and Cdc42p. However, the conditional enhanced density phenotype of ras2Δ cells is not a defect in secretion. All the phenotypes of ras2Δ cells can be fully suppressed by expression of yeast RAS1 or RAS2 genes, human Ha-ras, or the double disruption of the stress response genes msn2Δmsn4Δ. Although the best characterized pathway of Ras function in yeast involves activation of the cAMP-dependent protein kinase A pathway, activation of the protein kinase A pathway does not fully suppress the actin polarity defects, suggesting that there is an additional pathway from Ras2p to Msn2/4p. Thus, Ras2p regulates cytoskeletal polarity in yeast under conditions of mild temperature stress through the stress response pathway.

Cortisol fecal em ovinos: curva de excreção e estabilidade

O presente estudo foi dividido em dois experimentos, tendo como objetivo determinar a curva de excreção do cortisol fecal e sua estabilidade nas fezes perante exposição à diferentes períodos de tempo e temperatura entre as colheitas e análises, correlacionando os níveis de cortisol fecal com o pico de cortisol sanguíneo. No experimento 1, seis fêmeas mestiças (Dorper x Santa Inês) tiveram suas fezes totais colhidas durante 24 horas após a aplicação do hormônio adrenocorticotrófico (ACTH), além de colheitas de sangue realizadas antes da aplicação do ACTH, 60, 120 e 300 minutos depois; durante as quais foram atribuídos escores de reatividade para cada animal. Logo após as análises foi iniciado o experimento 2, no qual 9 cordeiros mestiços (Dorper x Santa Inês) foram submetidos a uma situação de estresse térmico durante os horários das 11 às 15 horas da tarde, tendo suas fezes colhidas às 23 horas do mesmo dia. Após a colheita, as fezes foram agrupadas e homogeneizadas em três grupos distintos, de onde retiraram-se alíquotas referentes aos tratamentos propostos: três temperaturas (15°, 25° e 35°) e quatro tempos (1, 3, 6 e 12 horas). Os dados da curva de excreção foram analisados por ANOVA, bem como pela correlação entre os valores de cortisol sanguíneo, fecal e reatividade. Para análise da estabilidade foi utilizada ANOVA multifatorial com dois fatores (temperatura e intervalo de tempo). Para avaliação das variáveis comportamentais foi realizada a transformação de escala dos dados para \"arco-seno raiz de porcentagem\", procedendo-se à análise de variância. O modelo estatístico contemplou os efeitos de dia (1, 2 e 3) com análise individual por animal. Os parâmetros de cortisol sanguíneo, frequência respiratória e temperatura retal foram analisados pelo teste t e correlação de Pearson. Todas as comparações de médias foram realizadas por teste F e teste t (PDIFF). A reatividade durante a colheita não exerceu efeito significativo sobre os valores de cortisol sanguíneo, os quais demonstraram médias maiores 60 minutos após a aplicação do ACTH e, após 300 minutos as ovelhas apresentaram níveis de cortisol considerados normais para ovinos sem estresse. Por outro lado, o pico de cortisol nas fezes foi verificado aproximadamente 10 a 12 horas após o pico de cortisol no sangue, não sendo verificadas diminuições significativas nas concentrações que indicassem o retorno aos níveis basais durante o período de 24 horas (P>0,05). Não foram observadas diferenças significativas entre os tempos e temperaturas aos quais as amostras de fezes foram submetidas (P>0,05), verificando-se uma tendência a manutenção da concentração do cortisol fecal em ovinos durante o período de 12 horas após a colheita.

Comportamento de Salmonella em ovo em pó em função da atividade de água (Aa) do binômio tempo x temperatura de armazenamento

Salmonella sp. é um dos principais microrganismos causadores de surtos de enfermidades transmitidas por alimentos associados ao consumo de ovos e de alimentos formulados com este ingrediente. Ovos desidratados são largamente utilizados pelas indústrias de alimentos, por oferecer maior praticidade e maior padronização em relação ao produto \"in natura\". Apesar do processo tecnológico de desidratação do ovo incluir uma etapa de pasteurização, existe um risco de haver microrganismos sobreviventes, já que a pasteurização é feita em temperatura branda. Além disso, a pasteurização pode destruir os fatores intrínsecos antimicrobianos presentes na clara, possibilitando a multiplicação de microrganismos que sobreviveram ao processo de pasteurização ou que contaminaram o produto após a pasteurização. O controle da Aa do produto desidratado e o tempo de armazenamento são, portanto, fatores fundamentais para o controle da multiplicação de microrganismos indesejáveis. Nesse estudo, avaliou-se a cinética de multiplicação de Salmonella experimentalmente adicionada a ovo em pó Aa ajustada para 0,4, 0,6, 0,8 e 0,9, durante o armazenamento em quatro temperaturas: 8°C, 15°C, 25°C e 35°C. Os resultados indicaram que S. Enteritidis é capaz de sobreviver por longo tempo (pelo menos 56 dias) em ovo em pó com Aa próximo de 0,4 quando armazenado a 8°C, 15° e a 25°C. Essa sobrevivência é menor (até 28 dias) quando o armazenamento é feito a 35°C. No ovo em pó com Aa em tomo de 0,6 ou 0,8, S. Enteritidis sobrevive por menos tempo do que no produto com Aa de cerca de 0,4, independentemente da temperatura de armazenamento. No produto com Aa de cerca de 0,9, há grande multiplicação de S. Enteritidis quando o armazenamento é feito a 15°C, 25°C ou 35°C. Nesse produto, o armazenamento a 8°C impede a multiplicação do patógeno. Verificou-se também que Salmonella Radar, resistente a diversos antibióticos, apresentou o mesmo comportamento que S. Enteritidis nas amostras de ovo estudadas.

Estudo cinético da lixiviação de metais de placas de circuito impresso obsoletas.

O processo tradicional de recuperação de metais de resíduos de equipamentos eletroeletrônicos (REEE) geralmente envolve processamento pirometalúrgico. Entretanto, o uso desta tecnologia para processar placas de circuito impresso (PCI) obsoletas pode levar à liberação de dioxinas e furanos, devido à decomposição térmica de retardantes de chama e resinas poliméricas presentes no substrato das placas. Portanto, este trabalho propõe uma rota hidrometalúrgica para recuperação de metais. O comportamento dos metais, com destaque para cobre, zinco e níquel, durante a lixiviação ácida, foi estudado em três temperaturas diferentes (35ºC, 65ºC e 75ºC), com e sem adição de um agente oxidante (peróxido de hidrogênio H2O2). A cinética de dissolução ácida desses metais foi estudada baseada na análise química por ICP-OES (Espectrometria de emissão ótica por plasma acoplado indutivamente) e EDX (Espectroscopia de fluorescência de raios-X por energia dispersiva). O balanço de massa e a análise química indicaram que a etapa de lixiviação sem adição de oxidante é pouco eficaz na extração dos metais, sendo responsável pela dissolução de menos do que 6% do total extraído. A 65ºC e H2SO4 1 mol/L, com adição de 5 mL de H2O2 (30%) a cada quinze minutos e densidade de polpa de 1 g / 10 mL, 98,1% do cobre, 99,9% do zinco e 99,0% do níquel foram extraídos após 4 horas. A cinética de dissolução desses metais é controlada pela etapa da reação química, seguindo, dependendo da temperatura, a equação 1 (1 XB)1/3 = k1.t ou a equação ln (1 XB) = k4.t.

Chlorophyll catabolism and gene expression in the peel of ripening banana fruits

The biochemical and molecular basis of chlorophyll (Chl) catabolism in bananas was investigated during ripening at 20°C and at an elevated temperature (35°C) where degreening is inhibited. Biochemical analysis showed that Chl breakdown products could be isolated from fruit ripened at both temperatures. The coloured breakdown products, chlorophyllide and pheophorbide, were not detected at any stage of ripening in the two treatments; however, a non-fluorescent Chl catabolite accumulated to a higher concentration at 20 than at 35°C. To investigate the ripening-related gene expression associated with these changes, a cDNA library was generated from the peel of fruit ripened at 20°C. Differential screening of this library produced 20 non-redundant families of clones including those encoding enzymes involved in ethylene biosynthesis, respiration, starch metabolism, cell wall degradation and other metabolic events. The expression of these genes was followed by northern analysis in fruit ripened at 20 and 35°C.

Preliminary evaluation of the common bean core collection at CIAT

Common bean (Phaseolus vulgaris L.), an annual diploid (2n=2x=22) species (Maréchal 1970; Delgado Salinas 1985), is adapted to mild temperatures (18°C to 35°C) and grown worldwide in a broad range of environments and in diverse production systems. Common bean is grown for its green leaves, green pods, and green and dry seeds. Dry leaves, threshed pods and stalks are fed to animals and used as fuel for cooking, especially in the developing countries of Africa and Asia (Singh 1991).

Pseudoperiodic "Living" and/or controlled cationic ring-opening copolymerization of oxetane with tetrahydropyran:microstructure of polymers vs kinetics of chain growth

The "living" and/or controlled cationic ring-opening bulk copolymerization of oxetane (Ox) with tetrahydropyran (THP) (cyclic ether with no homopolymerizability) at 35°C was examined using ethoxymethyl-1 -oxoniacyclohexane hexafluoroantimonate (EMOA) and (BF3 · CH3OH)THP as fast and slow initiator, respectively, yielding living and nonliving polymers with pseudoperiodic sequences (i.e., each pentamethylene oxide fragment inserted into the polymer is flanked by two trimethylene oxide fragments). Good control over number-average molecular weight (Mn up to 150000 g mol-1) with molecular weight distribution (MWD ∼ 1.4-1, 5) broader than predicted by the Poison distribution (MWDs > 1 +1/DPn) was attained using EMOA as initiating system, i.e., C 2H5OCH2Cl with 1.1 equiv of AgSbF6 as a stable catalyst and 1.1 equiv of 2,6-di-tert-butylpyridine used as a non-nucleophilic proton trap. With (BF3 · CH 3OH)THP, a drift of the linear dependence M n(GPC) vs Mn(theory) to lower molecular weight was observed together with the production of cyclic oligomers, ∼3-5% of the Ox consumed in THP against ∼30% in dichloromethane. Structural and kinetics studies highlighted a mechanism of chains growth where the rate of mutual conversion between "strain ACE species" (chain terminated by a tertiary 1-oxoniacyclobutane ion, Al) and "strain-free ACE species" (chain terminated by a tertiary 1-oxoniacyclohexane ion, Tl) depends on the rate at which Ox converts the stable species T1 (kind of "dormant" species) into a living "propagating" center A1 (i.e., k aapp[Ox]). The role of the THP solvent associated with the suspension of irreversible and reversible transfer reactions to polymer, when the polymerization is initiated with EMOA, was predicted by our kinetic considerations. The activation -deactivation pseudoequilibrium coefficient (Qt) was then calculated in a pure theoretical basis. From the measured apparent rate constant of Ox (kOxapp) and THP (kTHPapp = ka(endo)app) consumption, Qt and reactivity ratio (kp/kd, k a(endo)/ka(exo), and ks/ka(endo) were calculated, which then allow the determination of the transition rate constant of elementary step reactions that governs the increase of Mu with conversion. © 2009 American Chemical Society.

Geochemical composition of nodules and crusts from the Indian Ocean recovered from R/V Marion Dufresne expeditions from 1977 until 1981

Following the launch of the "Marion Dufresne 1", first supply ship of the Terres Australes and Antarctiques Françaises and part time oceanographic vessel in the Indian Ocean, a new marine geology program was developped at the Laboratoire de Géologie, MNHN. The first oceanographic cruise of the "Marion Dufresne 1" started in 1973 in the Southwestern Indian Ocean (OSIRIS I cruise). Forty piston-cores recovered nearly 200 m of sediments consisting in the first of the 450 cores of the Indian Ocean collection now deposited at the Museum. L. Leclaire being Director from 1980 to 1991, a multidisciplinary team (including sedimentologists and micropaleontologists) was involved in many oceanographic cruises in the Indian Ocean. Marine sedimentology was developped during annual cruises programs in collaboration with geophysicists, geochemists, and biologists. In 1995, the "Marion Dufresne 2" replaced the initial "Marion Dufresne 1".

Foraminíferos, ostracodes e microfauna associada da plataforma continental equatorial norte-rio-grandense, NE Brasil: área Porto do Mangue a Galinhos

This study was conducted in the adjacent Brazilian equatorial inner shelf to Rio Grande do Norte, between the region of Porto do Mangue and Galinhos. The main objective is the characterization of biogenic sediments, especially foraminifera and ostracod collected on the surface of the seafloor. The methodology involved standard procedures including literature, surveys, processing of samples in the laboratory and identification of foraminifera and ostracods by genera or species under stereo microscopy and scanning electron microscopy (SEM). Multivariate statistical analyzes and study of ecological indexes were also applied to the study of foraminifera. Three transects, from inner shelf to slope were sampled: profile 01 (east, near Galos), profile 02 (center, near the city of Macau) and profile 03 (west, near Ponta do Mel). Results indicated the predominance of benthic foraminifera and little plankton occurrence. Benthic foraminifera genera observed in abundance were Quinqueloculina, Textularia, Globigerina and Pyrgo, Quinqueloculina, Textularia, Pyrgo, Ammonia, Elphidium, Pseudononion, Peneroplis, Bolivina and Poroeponides, occurred more frequently. Less frequently been described Amphistegina, Archaias, Bigenerina, Cibicides, Cassidulina, Amphicorina, Cornuspira, Paterina, Hopkunsina, Oolina, Uvigerina, Fusenkoina, Nonionella, Amphisorus, Wiesrella, Reussella, Reophax, Nodosaria, Marginulina and Cyclogyra. Six genera of ostracods were also identified: Puriana variabilis / P. convoluted?, Loxoconcha sp, Bairdiidae, Xestoleberis sp, Hemicytheridae and Ruggiericythere sp. Groups of organisms found in the studied shelf presented chemical composition of Ca, C, O, Na, Cl, Al, Mg, and Si. The proportions of chemical elements may vary according to the type of biogenic sediment, with the highest values identified as Ca, C, Cl, Na and O. The absolute dating by carbon 14 method indicated sediments of different colors (light and dark), correspond to a single age from 3000 to 6000 years BP, related to the Quaternary. These data intend to complement information about biogenic sediments in the Brazilian continental shelf, especially in the Northeast, where there is a lack of such studies.

Foraminíferos, ostracodes e microfauna associada da plataforma continental equatorial norte-rio-grandense, NE Brasil: área Porto do Mangue a Galinhos

This study was conducted in the adjacent Brazilian equatorial inner shelf to Rio Grande do Norte, between the region of Porto do Mangue and Galinhos. The main objective is the characterization of biogenic sediments, especially foraminifera and ostracod collected on the surface of the seafloor. The methodology involved standard procedures including literature, surveys, processing of samples in the laboratory and identification of foraminifera and ostracods by genera or species under stereo microscopy and scanning electron microscopy (SEM). Multivariate statistical analyzes and study of ecological indexes were also applied to the study of foraminifera. Three transects, from inner shelf to slope were sampled: profile 01 (east, near Galos), profile 02 (center, near the city of Macau) and profile 03 (west, near Ponta do Mel). Results indicated the predominance of benthic foraminifera and little plankton occurrence. Benthic foraminifera genera observed in abundance were Quinqueloculina, Textularia, Globigerina and Pyrgo, Quinqueloculina, Textularia, Pyrgo, Ammonia, Elphidium, Pseudononion, Peneroplis, Bolivina and Poroeponides, occurred more frequently. Less frequently been described Amphistegina, Archaias, Bigenerina, Cibicides, Cassidulina, Amphicorina, Cornuspira, Paterina, Hopkunsina, Oolina, Uvigerina, Fusenkoina, Nonionella, Amphisorus, Wiesrella, Reussella, Reophax, Nodosaria, Marginulina and Cyclogyra. Six genera of ostracods were also identified: Puriana variabilis / P. convoluted?, Loxoconcha sp, Bairdiidae, Xestoleberis sp, Hemicytheridae and Ruggiericythere sp. Groups of organisms found in the studied shelf presented chemical composition of Ca, C, O, Na, Cl, Al, Mg, and Si. The proportions of chemical elements may vary according to the type of biogenic sediment, with the highest values identified as Ca, C, Cl, Na and O. The absolute dating by carbon 14 method indicated sediments of different colors (light and dark), correspond to a single age from 3000 to 6000 years BP, related to the Quaternary. These data intend to complement information about biogenic sediments in the Brazilian continental shelf, especially in the Northeast, where there is a lack of such studies.