The rate constant of photoinhibition, measured in lincomycin-treated leaves, is directly proportional to light intensity.

Pumpkin leaves grown under high light (500-700 micromol of photons m-2.s-1) were illuminated under photon flux densities ranging from 6.5 to 1500 micromol.m-2.s-1 in the presence of lincomycin, an inhibitor of chloroplast protein synthesis. The illumination at all light intensities caused photoinhibition, measured as a decrease in the ratio of variable to maximum fluorescence. Loss of photosystem II (PSII) electron transfer activity correlated with the decrease in the fluorescence ratio. The rate constant of photoinhibition, determined from first-order fits, was directly proportional to photon flux density at all light intensities studied. The fluorescence ratio did not decrease if the leaves were illuminated in low light in the absence of lincomycin or incubated in darkness in the presence of lincomycin. The constancy of the quantum yield of photoinhibition under different photon flux densities strongly suggests that photoinhibition in vivo occurs by one dominant mechanism under all light intensities. This mechanism probably is not the acceptor side mechanism characterized in the anaerobic case in vitro. Furthermore, there was an excellent correlation between the loss of PSII activity and the loss of the D1 protein from thylakoid membranes under low light. At low light, photoinhibition occurs so slowly that inactive PSII centers with the D1 protein waiting to be degraded do not accumulate. The kinetic agreement between D1 protein degradation and the inactivation of PSII indicates that the turnover of the D1 protein depends on photoinhibition under both low and high light.

Bacteriophage lambda N protein alone can induce transcription antitermination in vitro.

Specific and processive antitermination by bacteriophage lambda N protein in vivo and in vitro requires the participation of a large number of Escherichia coli proteins (Nus factors), as well as an RNA hairpin (boxB) within the nut site of the nascent transcript. In this study we show that efficient, though nonprocessive, antitermination can be induced by large concentrations of N alone, even in the absence of a nut site. By adding back individual components of the system, we also show that N with nut+ nascent RNA is much more effective in antitermination than is N alone. This effect is abolished if N is competed away from the nut+ RNA by adding, in trans, an excess of boxB RNA. The addition of NusA makes antitermination by the N-nut+ complex yet more effective. This NusA-dependent increase in antitermination is lost when delta nut transcripts are used. These results suggest the formation of a specific boxB RNA-N-NusA complex within the transcription complex. By assuming an equilibrium model, we estimate a binding constant of 5 x 10(6) M-1 for the interaction of N alone with the transcription complex. This value can be used to estimate a characteristic dissociation time of N from the complex that is comparable to the dwell time of the complex at an average template position, thus explaining the nonprocessivity of the antitermination effect induced by N alone. On this basis, the effective dissociation rate of N should be approximately 1000-fold slower from the minimally processive (100-600 bp) N-NusA-nut+ transcription complex and approximately 10(5)-fold slower from the maximally processive (thousands of base pairs) complex containing all of the components of the in vivo N-dependent antitermination system.

Phosphinate analogs of D-, D-dipeptides: slow-binding inhibition and proteolysis protection of VanX, a D-, D-dipeptidase required for vancomycin resistance in Enterococcus faecium.

VanX is a D-Ala-D-Ala dipeptidase that is essential for vancomycin resistance in Enterococcus faecium. Contrary to most proteases and peptidases, it prefers to hydrolyze the amino substrate but not the related kinetically and thermodynamically more favorable ester substrate D-Ala-D-lactate. The enzymatic activity of VanX was previously found to be inhibited by the phosphinate analogs of the proposed tetrahedral intermediate for hydrolysis of D-Ala-D-Ala. Here we report that such phosphinates are slow-binding inhibitors. D-3-[(1-Aminoethyl)phosphinyl]-D-2-methylpropionic acid I showed a time-dependent onset of inhibition of VanX and a time-dependent return to uninhibited steady-state rates upon dilution of the enzyme/inhibitor mixture. The initial inhibition constant Ki after immediate addition of VanX to phosphinate I to form the E-I complex is 1.5 microM but is then lowered by a relatively slow isomerization step to a second complex, E-I*, with a final K*i of 0.47 microM. This slow-binding inhibition reflects a Km/K*i ratio of 2900:1. The rate constant for the slow dissociation of complex E-I* is 0.24 min-1. A phosphinate analog with an ethyl group replacing what would be the side chain of the second D-alanyl residue in the normal tetrahedral adduct gives a K*i value of 90 nM. Partial proteolysis of VanX reveals two protease-sensitive loop regions that are protected by the intermediate analog phosphinate, indicating that they may be part of the VanX active site.

Dissociation between changes in cytoplasmic free Ca2+ concentration and insulin secretion as evidenced from measurements in mouse single pancreatic islets.

Simultaneous measurements of cytosolic free Ca2+ concentration and insulin release, in mouse single pancreatic islets, revealed a direct correlation only initially after stimulation with glucose or K+. Later, there is an apparent dissociation between these two parameters, with translocation of alpha and epsilon isoenzymes of protein kinase C to membranes and simultaneous desensitization of insulin release in response to glucose. Recovery of insulin release, without any concomitant changes in cytosolic free Ca2+ concentration, after addition of phorbol 12-myristate 13-acetate, okadaic acid, and forskolin supports the notion that the desensitization process is accounted for by dephosphorylation of key regulatory sites of the insulin exocytotic machinery.

A constant radius of curvature model for the organization of DNA in toroidal condensates.

Toroidal DNA condensates have received considerable attention for their possible relationship to the packaging of DNA in viruses and in general as a model of ordered DNA condensation. A spool-like model has primarily been supported for DNA organization within toroids. However, our observations suggest that the actual organization may be considerably different. We present an alternate model in which DNA for a given toroid is organized within a series of equally sized contiguous loops that precess about the toroid axis. A related model for the toroid formation process is also presented. This kinetic model predicts a distribution of toroid sizes for DNA condensed from solution that is in good agreement with experimental data.

Basis of guanylate cyclase activation by carbon monoxide.

Kinetics of CO association with guanylate cyclase [GTP pyrophosphate-lyase (cyclizing), EC 4.6.1.2] and dissociation from carboxy guanylate cyclase have been studied at pH 7.5 by flash photolysis, yielding rate constants at 23 degrees C of 1.2 +/- 0.1 x 10(5) M-1.sec-1 and 28 +/- 2 sec-1, respectively. While the CO combination rate constant is the same as for the T state of hemoglobin, the CO dissociation rate constant is much higher than expected for a six-coordinate carboxyheme protein; yet the absorption spectrum is indicative of a six-coordinate heme. The two observations are reconciled by a reaction mechanism in which CO dissociation proceeds via a five-coordinate intermediate. This intermediate is structurally very similar to the five-coordinate nitrosyl heme derivative of guanylate cyclase and is presumably responsible for the observed 4-fold activation of guanylate cyclase by CO. Thus, we provide a model that explains enzyme activities of the nitrosyl and carboxy forms of the enzyme on the basis of a common mechanism.

Chemical Reactors. Problem 2: Constant pressure adiabatic stirred batch reactor with variable heat capacities (2011-2012 Course)

Wording of problem 2 (week 3, 17/10/11).

Chemical Reactors. Problem 2: Constant pressure adiabatic stirred batch reactor with variable heat capacities. Solution (2011-2012 Course)

Chemical Reaction Engineering. Course 2011-12. Solution of problem 2: constant pressure adiabatic stirred batch reactor with variable heat capacities.

Chloride Penetration Prediction in Concrete through an Empirical Model Based on Constant Flux Diffusion

An empirical model based on constant flux is presented for chloride transport through concrete in atmospherical exposure conditions. A continuous supply of chlorides is assumed as a constant mass flux at the exposed concrete surface. The model is applied to experimental chloride profiles obtained from a real marine structure, and results are compared with the classical error-function model. The proposed model shows some advantages. It yields a better predictive capacity than the classical error-function model. The previously observed chloride surface concentration increases are compatible with the proposed model. Nevertheless, the predictive capacity of the model can fail if the concrete microstructure changes with time. The model seems to be appropriate for well-maturated concretes exposed to a marine environment in atmospherical conditions.

Dynamical Equations For The Period Vectors In A Periodic System Under Constant External Stress

The purpose of this paper is to derive the dynamical equations for the period vectors of a periodic system under constant external stress. The explicit starting point is Newton’s second law applied to halves of the system. Later statistics over indistinguishable translated states and forces associated with transport of momentum are applied to the resulting dynamical equations. In the final expressions, the period vectors are driven by the imbalance between internal and external stresses. The internal stress is shown to have both full interaction and kinetic-energy terms.

Les effets de l’éclairage cyclique versus l’éclairage tamisé constant sur la stabilité physiologique et le niveau d’activité motrice de prématurés

Problématique : Les prématurés évoluent dans l’unité néonatale qui présente une intensité lumineuse parfois forte et variable, ce qui a pour effet de provoquer une instabilité physiologique, ainsi qu’une augmentation du niveau d’activité motrice chez ces derniers. Par ailleurs, le contrôle de l’éclairage à l’unité néonatale favorise la stabilité physiologique et réduit le niveau d’activité motrice des prématurés. Deux méthodes de contrôle de l’éclairage ont été étudiées, soit l’éclairage tamisé constant et l’éclairage cyclique. Or, la méthode de contrôle de l’éclairage la plus appropriée au système nerveux immature des prématurés est inconnue et il y a ambivalence en ce qui concerne les résultats des études ayant évalué ces deux modes de contrôle de l’éclairage. But : Le but de cette étude était de mesurer les effets de l’éclairage cyclique versus l’éclairage tamisé constant sur la stabilité physiologique et le niveau d’activité motrice de prématurés nés entre 28 et 32 semaines d’âge gestationnel. Méthode : Un essai clinique randomisé a été réalisé. Les 38 prématurés recrutés dans une unité néonatale de niveaux II et III d’un hôpital universitaire, ont été randomisés dans l’un des deux groupes d’intervention, soit le groupe exposé à l’éclairage tamisé constant ou celui exposé à l’éclairage cyclique. Ces deux types d’éclairage ont été appliqués pendant 24 heures. La stabilité physiologique a été mesurée par le score Stability of the Cardio Respiratory System in Premature Infants (SCRIP) et le niveau d’activité motrice a été mesuré avec un accéléromètre (Actiwatch®). L’intensité lumineuse à laquelle les prématurés ont été exposés a été mesurée de façon continue à l’intérieur de l’incubateur à l’aide d’un photomètre. Résultats : L’analyse des données révèle qu’il n’y aucune différence significative entre les deux groupes d’intervention en ce qui a trait à la stabilité physiologique (valeur-p du score SCRIP de 0,54 à 0,96) et au niveau d’activité motrice (valeur-p de 0,09 à 0,88). Les participants des deux groupes ont manifesté une stabilité physiologique et un niveau d’activité motrice comparables. Conclusion : Des interventions de contrôle de l’éclairage doivent être adoptées à l’unité néonatale, que ce soit des interventions qui permettent la mise en œuvre de l’éclairage cyclique ou de l’éclairage tamisé constant, dans le but de favoriser l’adaptation du prématuré à l’environnement de l’unité néonatale. Des recherches additionnelles sont requises afin d’identifier la méthode de contrôle de l’éclairage (éclairage cyclique ou éclairage tamisé constant) qui doit être implantée à l’unité néonatale.

Twenty-eight national elections for a Parliament in constant evolution. European Policy Brief No. 26, March 2014

In the City, the citizen is king. At least theoretically. In the European City currently being built around twenty-eight national democracies, the citizen will soon be called upon, in May, to democratically elect his or her representative in the European Parliament for the next five years. Since the very first election of Members of the European Parliament by direct universal suffrage in 1979, spectacular progress has been made by the “European Economic Community” that we now all know as the European Union. And the powers vested in citizen representatives are equally impressive. But there is a real possibility that European citizens will turn their backs on the upcoming European elections like never before. Why?