Resposta do Feijoeiro Comum à Inoculação com Rizóbio e Suplementação com Nitrogênio Mineral em Dois Biomas Brasileiros

O feijoeiro comum (Phaseolus vulgaris L.) pode se beneficiar da fixação biológica de N2, mas respostas inconsistentes da cultura à inoculação com rizóbio indicam a necessidade de aplicação de N mineral complementar. Este trabalho teve por objetivo avaliar a resposta do feijoeiro à inoculação com rizóbio, associada à suplementação com N mineral, nos biomas Cerrado e Mata Atlântica. Foram conduzidos quatro experimentos de campo, dois em Santo Antônio de Goiás, GO, um em Valença, RJ e um em Macaé, RJ, onde a inoculação com estirpes comerciais de rizóbio foi comparada à inoculação com a estirpe BR 923 de Sinorhizobium sp., à adubação com N mineral e à suplementação com N na semeadura e em cobertura. A avaliação da população nativa de rizóbio indicou 105 células g-1 no solo na área experimental em Goiás, anteriormente cultivada com feijão, e 102células g-1 em Valença, anteriormente mantida com pastagem. Nos dois experimentos em Goiás, o rendimento de grãos, da ordem de 2.100 kg ha-1, não diferiu entre os tratamentos testemunha absoluta, inoculação com rizóbio ou aplicação de 120 kg ha-1 de N. Em Valença, a inoculação com estirpes comerciais forneceu rendimentos da cultivar Ouro Negro superiores à testemunha absoluta, na ausência de adubação de cobertura; na presença de 40 kg ha-1 de N em cobertura, a inoculação com rizóbio proporcionou rendimento de 3.420 kg ha-1, superior aos demais tratamentos. Na média das diferentes fontes de N na semeadura, a adubação de cobertura aumentou a produção de grãos de 2.367 para 2.542 kg ha-1. Em Macaé, em solo com alto teor de matéria orgânica, os maiores rendimentos foram obtidos com inoculação das estirpes comerciais associada a 40 kg ha-1 de N em cobertura, com efeito deletério da adubação de 80 kg ha-1 de N no plantio. Concluiu-se que em áreas sem cultivo prévio de feijão, a inoculação com estirpes comerciais de rizóbio aumentou o rendimento de grãos, em particular quando associada à adubação de cobertura com N.

Expression, purification, and structural insights for the human uric acid transporter, GLUT9, using the Xenopus laevis oocytes system.

The urate transporter, GLUT9, is responsible for the basolateral transport of urate in the proximal tubule of human kidneys and in the placenta, playing a central role in uric acid homeostasis. GLUT9 shares the least homology with other members of the glucose transporter family, especially with the glucose transporting members GLUT1-4 and is the only member of the GLUT family to transport urate. The recently published high-resolution structure of XylE, a bacterial D-xylose transporting homologue, yields new insights into the structural foundation of this GLUT family of proteins. While this represents a huge milestone, it is unclear if human GLUT9 can benefit from this advancement through subsequent structural based targeting and mutagenesis. Little progress has been made toward understanding the mechanism of GLUT9 since its discovery in 2000. Before work can begin on resolving the mechanisms of urate transport we must determine methods to express, purify and analyze hGLUT9 using a model system adept in expressing human membrane proteins. Here, we describe the surface expression, purification and isolation of monomeric protein, and functional analysis of recombinant hGLUT9 using the Xenopus laevis oocyte system. In addition, we generated a new homology-based high-resolution model of hGLUT9 from the XylE crystal structure and utilized our purified protein to generate a low-resolution single particle reconstruction. Interestingly, we demonstrate that the functional protein extracted from the Xenopus system fits well with the homology-based model allowing us to generate the predicted urate-binding pocket and pave a path for subsequent mutagenesis and structure-function studies.

Severely presenting strokes : profile & outcome

Etat des connaissances: Les Accidents vasculaires cérébraux (AVC) sévères sont une cause importante de mortalité et de handicap en Suisse. Les buts de cette étude sont de déterminer les caractéristiques des patients avec un AVC à présentation initiale sévère "severely presenting", ainsi que d'identifier les facteurs prédicteurs en phase aigüe et subaigüe d'un devenir favorable chez ces patients. Methodologie: En utilisant le registre des AVC "Acute STroke Registry and Analysis of Lausanne (ASTRAL), nous avons comparé tous les patients avec un AVC "à présentation sévère", defini comme un score National Institute of Health Stroke Scale (NIHSS) > 20 à l'admission aux urgences, avec tous les autres patients du registre. Dans une analyse statistique multivariée, les associations avec les caractéristiques démographiques, cliniques, pathophysiologiques, métaboliques et radiologiques des patients on été déterminées. Dans un deuxième temps, nous avons analysé les facteurs prédicteurs d'un devenir favorable à 3 mois (modified Rankin scale (mRS) <3) dans ce groupe d'AVC à présentation sévère. Resultats: Parmi les 1 '915 patients consécutifs, 243 (12.7%) présentaient un AVC rejoignant la définition de sévère. Ceux-cis étaient associés de manière significative avec un méchanisme ischémique cardio-embolique (OR=1.74 / 95% CI 1.19 - 2.54), un début inconnu de la Symptomatologie (OR=2.35 / 95% CI 1.14 - 4.83), avaient plus de trouvailles d'origine ischémique aigüe lors de l'imagerie dJentrée (la majorité sont des CT-scan, OR=2.65 / 95% CI 1.79 - 3.92), plus d'occlusions dans l'imagerie vasculaire d'admission (OR=27.01 / 95% CI 11.5 - 62.9), moins d'anciens infarctus cérébraux sur l'imagerie (OR=0.43 / 95% 0.26¬0.72), un taux d'haemoglobine plus bas en g/1 (OR=0.97 / 95% CI 0.96 - 0.99), un taux de leucocytes plus élevé par 1000 cells/1 (OR=1.05 / 95%CI 1.00 - 1.11). parmi les 68 (28%) patients avec un devenir favorable malgré un AVC initialement sévère, leur évolution favorable à été associée avec un âge plus jeune (OR=0.94 / 95% CI 0.92 - 0.97), la présence d'évenements cérébrovasculaires antécédants (OR=3.00 / 95% CI 1.01 - 8.97), un traitement hypolipémiant déjà présent (OR= 3.82 / 95% CI 1.34 - 10.90), une température corporelle d'admission plus basse (0R=0.43 1 95% CI 0.23 - 0.78), une concentration subaigüe de glucose plus basse (OR=0.74 1 95% CI 0.56 - 0.97), et une recanalisation spontanée ou par thrombolyse à 24h (OR= 4.51 1 95%CI 1.96- 10.41). Conclusion: les AVC à présentation initiale severe sont associés à des facteurs prédicteurs cliniques, radiologiques, et métaboliques multiples, dont certains sont modifiables. Les facteurs prédicteurs des 28% de patients avec un devenir favorable en dépit d'un AVC intitialement sévère sont un pré-traitement par hypolipémiants, une temperature corporelle plus basse à l'admission, une glycémie plus basse à 24heures et la recanalisation artérielle.

Analysis of the mechanisms controlling the activity of flp1, a chromosomal passenger protein in the fission Schizosaccharomyces pombe

ABSTRACT In S. cerevisiae, the protein phosphatase Cdc14pwt is essential far mitotic exit through its contribution to reducing mitotic CDK activity. But Cdc14pwt also acts as a mare general temporal coordinator of mid and late mitotic events by controlling the partitioning of DNA, microtubule stability and cytokinesis. Cdc14pwt orthologs are well conserved from yeasts to humans, and sequence comparison revealed the presence of three domains, A, B and C, of which A and B form the catalytic domain. Cdc14pwt orthologs are regulated (in part) through cell cycle dependent changes in their localization. Some of them are thought to be kept inactive by sequestration in the nucleolus during interphase. This is the case for flp1pwt, the single identified Cdc14pwt ortholog in the fission yeast S. pombe. In early mitosis, flp1pwt leaves the nucleolus and localizes to the kinetochores, the contractile ring and the mitotic spindle, suggesting that it has multiple substrates and regulates many mitotic processes. flp1D cells show a high chromosome loss rate and septation defects, suggesting a role for flp1wt in the fidelity of chromosome transmission and cytokinesis. The aim of this study is to characterize the mechanisms underlying flp1pwt functions and the control of its activity. A structure-function analysis has revealed that the presence of both A and B domains is required for biological function and for proper flp1pwt mitotic localization. In contrast, the C domain of flp1pwt is responsible for its proper nucleolar localization in G2/interphase. My data suggest that dephosphorylation of substrates by flp1pwt is not necessary for any changes in localization of flp1pwt except that at the medial ring. In that particular case, the catalytic activity of flp1pwt is required for efficient localization, therefore revealing an additional level of regulation. All the functions of flp1pwt assayed to date require its catalytic activity, emphasizing the importance of further identification of its substrates. As described for other orthologs, the capability of selfinteraction and phosphorylation status might help to control flp1pwt activity. My data suggest that flp1pwt forms oligomers in vivo and that phosphorylation is not essential far localization changes of the protein. In addition, the hypophosphorylated form of flp1pwt might be specifically involved in the promotion of cytokinesis. The results of this study suggest that multiple modes of regulation including localization, selfassociation and phosphorylation allow a fine-tuning regulation of flp1pwt phosphatase activity, and more generally that of Cdc14pwt family of phosphatases. RESUME Chez la levure S. cerevisiae, la protéine phosphatase Cdc14pwt est essentielle pour la sortie de mitose du fait de sa contribution dans la réduction d'activité des CDK mitotiques. Comme elle contrôle également le partage de l'ADN, la stabilité des microtubules et la cytokinèse, Cdc14pwt est en fait considérée comme un coordinateur temporel général des évènements de milieu et de fin de mitose. Les orthologues de Cdc14pwt sont bien conservés, des levures jusqu'à l'espèce humaine. Des comparaisons de séquence ont révélé la présence de trois domaines A, B et C, les deux premiers constituant le domaine catalytique. Ils sont régulés (en partie) via des changements dans leur localisation, eux-mêmes dépendants du cycle cellulaire. Plusieurs de ces orthologues sont supposés inactivés par séquestration dans le nucléole en interphase, ce qui est le cas de flp1pwt le seul orthologue de Cdc14pwt identifié chez la levure fissipare S, pombe. En début de mitose, flp1pwt quitte le nucléole et localise au niveau des kinetochores, de l'anneau contractile d'actine et du fuseau mitotique, ce qui laisse supposer de multiples substrats et fonctions. Comme les cellules délétées pour le gène flp1wt présentent un taux élevé de perte de chromosome et des défauts de septation, flp1pwt semble jouer un rôle dans la fidélité de la transmission du matériel génétique et la cytokinèse. Le but de cette étude est de caractériser les mécanismes impliqués dans les fonctions assurées par flp1pwt d'une part, et dans le contrôle de son activité d'autre part. Une analyse structure-fonction a révélé que la présence simultanée des deux domaines A et B est requise pour la fonction biologique de flp1pwt et sa localisation correcte pendant la mitose. Par contre, le domaine C de flp1pwt confère une localisation nucléolaire adéquate en G2/interphase. Mes données suggèrent que la déphosphorylation de substrats par flp1pwt est dispensable pour sa localisation correcte excepté celle à l'anneau médian, qui requiert dans ce cas, l'activité catalytique de flp1pwt, révélant ainsi un niveau de régulation supplémentaire. Toutes les fonctions de flp1 pwt testées jusqu'à présent nécessitent également son activité catalytique, ce qui accentue l'importance de l'identification future de ses substrats. Comme cela a déjà été décrit pour d'autres orthologues, la capacité d'auto-intéraction et le niveau de phosphorylation pourraient contrôler l'activité de flp1pwt. En effet, mes données suggèrent que flp1pwt forme des oligomères in vivo et que la phosphorylation n'est pas essentielle pour les changements de localisation observés pour la protéine. De plus, la forme hypophosphorylée de flp1pwt pourrait être spécifiquement impliquée dans la promotion de la cytokinèse. De multiples modes de régulation incluant la localisation, l'auto-association et la phosphorylation semblent permettre un contrôle fin et subtil de l'activité de la phosphatase flp1pwt, et plus généralement celle des protéines de la famille de Cdc14pwt.

A general approach to the in situ energy butget of Eudendrium racemosum (Cnidaria, Hydrozoa) in the Western Mediterranean

An in situ energy budget of the hydropolyp Eudendrium racemosum (Cavolini, 1785) is presented. Ingestion and respiration rates and ammonium excretion were studied over two 24 h cycles, with two-hour sample intervals. The species ingested as much as 25.9% of its own biomass per day (minimum rate). Respiration was 1.62 ml O2 g-1 d w h-1 while excretion was 13.6 mM NH4 g-1dw h-1. We estimated that the species increased its biomass at a rate of 9.6% per day (Growth + Reproduction). This value is higher than those previously reported for other cnidarians. We can assume that the capacity of E. racemosum to survive - albeit for a limited period of the year - in the highly-competitive shallow-water communities is based on its high growth rate.

La main de velours / par Mlle Gabrielle d'Éthampes

Collection : Collection D. S. G.

Effects of nonhypotensive endotoxemia in conscious rats: role of prostaglandins.

A nonhypotensive dose of endotoxin was administered to normal conscious rats to evaluate the vascular and humoral effects of endotoxemia per se. Mean blood pressure and heart rate remained stable during the 45 min infusion of Escherichia coli endotoxin (0.01 mg/min). However, a marked increase in plasma renin activity (4.2 +/- 0.48 vs. 30.2 +/- 6 ng.ml-1.h-1, mean +/- SE, P less than 0.01), plasma epinephrine (0.112 +/- 0.04 vs. 1.71 +/- 0.5 ng/ml, P less than 0.01), and plasma norepinephrine (0.269 +/- 0.028 vs. 1.3 +/- 0.2 ng/ml, P less than 0.001) was observed during infusion in endotoxin-treated rats when compared with the vehicle-treated animals. In addition, the blood pressure response to exogenous norepinephrine was significantly reduced during nonhypotensive endotoxemia. Significant changes in regional blood flow distribution, as assessed by radiolabeled microspheres, were observed in endotoxemic rats; in particular a decrease in renal blood flow (7.39 +/- 0.43 vs. 5.97 +/- 0.4 ml.min-1.g-1, P less than 0.05) and an increase in coronary blood flow (5.01 +/- 0.38 vs. 6.44 +/- 0.33 ml.min-1.g-1, P less than 0.01) were found. The role of prostaglandins in the vascular and humoral alterations induced by nonhypotensive endotoxemia was also examined. Pretreatment with indomethacin (5 mg) prevented the increase in plasma renin activity as well as plasma catecholamine levels. On the contrary, the decreased vascular reactivity and the reduction in renal blood flow observed during endotoxemia were not affected by prostaglandin synthesis inhibition. Thus significant vascular and humoral changes have been found during endotoxemia even in absence of hypotension.(ABSTRACT TRUNCATED AT 250 WORDS)

Linker insertion mutagenesis based on IS21 transposition: isolation of an AMP-insensitive variant of catabolic ornithine carbamoyltransferase from Pseudomonas aeruginosa.

The bacterial insertion sequence IS21 when repeated in tandem efficiently promotes non-replicative cointegrate formation in Escherichia coli. An IS21-IS21 junction region which had been engineered to contain unique SalI and BglII sites close to the IS21 termini was not affected in the ability to form cointegrates with target plasmids. Based on this finding, a novel procedure of random linker insertion mutagenesis was devised. Suicide plasmids containing the engineered junction region (pME5 and pME6) formed cointegrates with target plasmids in an E.coli host strain expressing the IS21 transposition proteins in trans. Cointegrates were resolved in vitro by restriction with SalI or BglII and ligation; thus, insertions of four or 11 codons, respectively, were created in the target DNA, practically at random. The cloned Pseudomonas aeruginosa arcB gene encoding catabolic ornithine carbamoyltransferase was used as a target. Of 20 different four-codon insertions in arcB, 11 inactivated the enzyme. Among the remaining nine insertion mutants which retained enzyme activity, three enzyme variants had reduced affinity for the substrate ornithine and one had lost recognition of the allosteric activator AMP. The linker insertions obtained illustrate the usefulness of the method in the analysis of structure-function relationships of proteins.

Parâmetros microbiológicos indicadores do efeito do diuron sobre a microflora do solo

A biomassa microbiana (Cmic), a respiração básica (RB), as razões Cmic:Corg e o qCO2 foram usados para avaliar o efeito do herbicida diuron, nas dosagens de 0, 2 e 20 mig g-1 , na microflora do solo. O estudo foi conduzido em dois solos com texturas diferentes, e as avaliações, realizadas aos 0, 3, 10, 20 e 33 dias após a aplicação do pesticida. No solo mais argiloso, o Cmic e a razão Cmic:Corg forneceram resultados semelhantes quanto ao efeito prejudicial do diuron. A RB não foi afetada pelo diuron. O qCO2 foi o único parâmetro capaz de demonstrar uma diferenciação no efeito prejudicial entre doses. No solo mais arenoso não se detectou efeito do diuron.

Crescimento e resistência à seca de leucena em solo de cerrado

Plantas envasadas de Leucaena leucocephala Lam. (De Witt) foram cultivadas em casa de vegetação sob 30% de sombreamento artificial, em solo de cerrado, com adição de 0, 750, 1.500 e 3.000 kg ha-1 de NPK (4-14-8). Aos 180 dias após a emergência (DAE), as plantas crescidas em solo sem adubo químico apresentaram os menores valores de taxa de crescimento relativo (TCR = 0,028 g g-1 dia-1), biomassa total (5,2 g), taxa assimilatória líquida (TAL= 4,2.10-4 g cm-2 dia-1), e não produziram nem flores nem vagens. Entre as plantas cultivadas em solo fertilizado, as que receberam maiores quantidades de adubo químico apresentaram maiores valores de biomassa total e área foliar, a partir de 90 DAE. Porém, os maiores valores de TCR (43,92.10-4 g g-1 dia-1) e TAL (15,01 g cm-2 dia-1) foram obtidos nas plantas crescidas em solo com 750 kg ha-1 de NPK. A presença de flores e vagens nas plantas que receberam 1.500 e 3.000 kg ha-1 de NPK ocorreu a partir dos 120 DAE, e aos 150 DAE se iniciou o processo de abscisão foliar nos vegetais crescidos em todos os tratamentos, com conseqüente redução da área foliar, TAL e TCR. Na presença de estresse hídrico simulado, as plantas com 90 DAE mostraram-se mais resistentes que as de 30 dias. Porém, nos dois casos, as plantas mais tolerantes à dessecação foram as crescidas em solo com maiores teores de NPK, e um ajuste osmótico auxiliou as plantas a sobreviverem durante o período da seca.

PERSISTENCE OF ATRAZINE, METRIBUZIN AND SIMAZINE HERBICIDES IN TWO SOILS

An oat bioassay was conducted in pots under greenhouse conditions to determine the persistence of atrazine, metribuzin and simazine herbicides in soils of the southeast of Buenos Aires Province, Argentina. Atrazine rates of 0, 0.58, 1.16 and 2.32 mug g-1 of active ingredient (a.i.), metribuzin rates of 0, 0.14, 0.28 and 0.56 mug g-1 of a.i., and simazine rates of 0, 0.72, 1.45 and 2.9 mug g-1 of a.i. dry soil weight were applied to pots containing soils from Balcarce and San Cayetano sites. Organic matter (OM) content and pH of Balcarce soil were 5.5% and 5.8%, while for San Cayetano soil were 2.9% and 6.7%, respectively. Relative dry weight (RDW) of oat shoots was calculated as percentage of control. Considering a 20% RDW reduction of oat shoots, persistences of recommended rates for the region were: atrazine (1.16 mug g-1 of a.i.), 78 and 130 days after treatment (DAT) for Balcarce and San Cayetano soil, respectively; metribuzin (0.28 mug-1 of a.i.), 63 and 77 DAT for Balcarce and San Cayetano soil, respectively; simazine (1.45 mug g-1 of a.i.), 81 and 156 DAT for Balcarce and San Cayetano soil, respectively. Results show that persistence of atrazine, metribuzin and simazine in soil increased with high rates, low OM content and high pH.

Evolvimento de dióxido de carbono e mineralização de nitrogênio em Latossolo Vermelho-Escuro com diferentes manejos

Este trabalho foi elaborado com o objetivo de verificar o efeito das palhas de milho e lab-lab, dos tipos de manejos do solo e da presença de uma fonte externa de N, no evolvimento de CO2 e na mineralização de N. Usou-se um Latossolo Vermelho-Escuro distrófico textura argilosa fase cerrado, da região de Sete Lagoas (MG), com histórico de uso envolvendo cinco anos sob plantio direto e plantio convencional com arado de disco e arado de aiveca. Amostras de solo (0-20 cm) foram incubadas a 25ºC durante 55 dias, na presença e na ausência de palhada residual e de fonte externa de N aplicada no início e 25 dias após. De modo geral, a palhada de milho apresentou maior taxa de CO2 evolvido do que o lab-lab (935 e 764 mig CO2 g-1 solo, respectivamente). O tipo de palhada residual influenciou mais o evolvimento de CO2 do que os manejos de solo. A adição de N aumenta a taxa de CO2 evolvida, apenas quando N é aplicado no início do período de incubação. A disponibilidade de N e o tempo de imobilização são alterados pela relação C/N da palhada incorporada e pelo preparo do solo.

Efeito da suplementação orgânica sobre a toxidez do fungicida clorotalonil na microbiota do solo

Avaliou-se o efeito da suplementação do solo com material orgânico (aveia nas proporções de 0, 7,5 e 15 g kg-1), sobre a toxidez do fungicida clorotalonil (0, 12 e 48 µg de i.a. g-1 de solo), na microbiota do solo. As avaliações ocorridas aos 11 e 26 dias após esta aplicação demonstraram que, embora tenha ocorrido uma recuperação do carbono da biomassa microbiana, as doses de aveia utilizadas ainda não foram suficientes para inibir o efeito adverso do fungicida. Os resultados de comprimento de hifas vivas demonstraram que não foi possível recuperar a população fúngica do solo na maior dose do fungicida.

Adsorption of glyphosate in chilean soils and its relationship with unoccupied phosphate binding sites

The objective of this work was to investigate glyphosate adsorption by soils and its relationship with unoccupied binding sites for phosphate adsorption. Soil samples of three Chilean soils series - Valdivia (Andisol), Clarillo (Inceptisol) and Chicureo (Vertisol) - were incubated with different herbicide concentrations. Glyphosate remaining in solution was determined by adjusting a HPLC method with a UV detector. Experimental maximum adsorption capacity were 15,000, 14,300 and 4,700 mg g¹ for Valdivia, Clarillo, and Chicureo soils, respectively. Linear, Freundlich, and Langmuir models were used to describe glyphosate adsorption. Isotherms describing glyphosate adsorption differed among soils. Maximum adjusted adsorption capacity with the Langmuir model was 231,884, 17,874 and 5,670 mg g-1 for Valdivia, Clarillo, and Chicureo soils, respectively. Glyphosate adsorption on the Valdivia soil showed a linear behavior at the range of concentrations used and none of the adjusted models became asymptotic. The high glyphosate adsorption capacity of the Valdivia soil was probably a result of its high exchangeable Al, extractable Fe, and alophan and imogolite clay type. Adsorption was very much related to phosphate dynamics in the Valdivia soil, which showed the larger unoccupied phosphate binding sites. However relationship between unoccupied phosphate binding sites and glyphosate adsorption in the other two soils (Clarillo and Chicureo) was not clear.

Basil conservation affected by cropping season, harvest time and storage period

Fresh basil (Ocimum basilicum L.) is used in food, phytotherapic industry, and in traditional therapeutic, due to its essential oil content and composition. Nevertheless basil can not be kept for long periods after harvest and its quality can be reduced. This work aimed to assess the influence of the season and harvest time in the postharvest conservation of basil stored for different periods. Basil was harvested at 8 am and 4 pm both in August/1999 and January/2000. Cuttings were conditioned in PVC packages and stored for 3, 6, and 9 days. During storage, chlorophyll content, essential oil content and composition were determined as well as microbiological analyses were carried out. Harvest season and the days of storage influenced the final content of essential oil. There was a linear decrease in the content of essential oil, in the chlorophyll content and in the number of mold and yeast colonies during storage. There was no effect of cropping season or harvest hour on essential oil composition, but the eugenol and linalool content increased during storage. Coliforms were under 0.3 MPN g-1 and the number of Staphylococcus aureus was under 1.0x10² UFC g-1.