An ensemble and single-molecule fluorescence microscopy investigation of phase-separated monolayer films stained with Nile Red

Phase-separated Langmuir-Blodgett monolayer films prepared from mixtures of arachidic acid (C19H39COOH) and perfluorotetradecanoic acid (C13F27COOH) were stained via spin-casting with the polarity sensitive phenoxazine dye Nile Red, and characterized using a combination of ensemble and single-molecule fluorescence microscopy measurements. Ensemble fluorescence microscopy and spectromicroscopy showed that Nile Red preferentially associated with the hydrogenated domains of the phase-separated films, and was strongly fluorescent in these areas of the film. These measurements, in conjunction with single-molecule fluorescence imaging experiments, also indicated that a small sub-population of dye molecules localizes on the perfluorinated regions of the sample, but that this sub-population is spectroscopically indistinguishable from that associated with the hydrogenated domains. The relative importance of selective dye adsorption and local polarity sensitivity of Nile Red for staining applications in phase-separated LB films as well as in cellular environments is discussed in context of the experimental results.     

The role of the C8 proton of ATP in the catalysis of shikimate kinase and adenylate kinase

ABSTRACT: BACKGROUND: It has been demonstrated that the adenyl moiety of ATP plays a direct role in the regulation of ATP binding and/or phosphoryl transfer within a range of kinase and synthetase enzymes. The role of the C8-H of ATP in the binding and/or phosphoryl transfer on the enzyme activity of a number of kinase and synthetase enzymes has been elucidated. The intrinsic catalysis rate mediated by each kinase enzyme is complex, yielding apparent KM values ranging from less than 0.4 muM to more than 1 mM for ATP in the various kinases. Using a combination of ATP deuterated at the C8 position (C8D-ATP) as a molecular probe with site directed mutagenesis (SDM) of conserved amino acid residues in shikimate kinase and adenylate kinase active sites, we have elucidated a mechanism by which the ATP C8-H is induced to be labile in the broader kinase family. We have demonstrated the direct role of the C8-H in the rate of ATP consumption, and the direct role played by conserved Thr residues interacting with the C8-H. The mechanism by which the vast range in KM might be achieved is also suggested by these findings. RESULTS: We have demonstrated the mechanism by which the enzyme activities of Group 2 kinases, shikimate kinase (SK) and adenylate kinase 1 (AK1), are controlled by the C8-H of ATP. Mutations of the conserved threonine residues associated with the labile C8-H cause the enzymes to lose their saturation kinetics over the concentration range tested. The relationship between the role C8-H of ATP in the reaction mechanism and the ATP concentration as they influence the saturation kinetics of the enzyme activity is also shown. The SDM clearly identified the amino acid residues involved in both the catalysis and regulation of phosphoryl transfer in SK and AK1 as mediated by C8H-ATP. CONCLUSIONS: The data outlined serves to demonstrate the "push" mechanism associated with the control of the saturation kinetics of Group 2 kinases mediated by ATP C8-H. It is therefore conceivable that kinase enzymes achieve the observed 2,500-fold variation in KM through a combination of the various conserved "push" and "pull" mechanisms associated with the release of C8-H, the proton transfer cascades unique to the class of kinase in question and the resultant/concomitant creation of a pentavalent species from the gamma-phosphate group of ATP. Also demonstrated is the interplay between the role of the C8-H of ATP and the ATP concentration in the observed enzyme activity. The lability of the C8-H mediated by active site residues co-ordinated to the purine ring of ATP therefore plays a significant role in explaining the broad KM range associated with kinase steady state enzyme activities.     

Magnetic resonance imaging of intraductal papilloma of the breast

To describe the appearance of isolated intraductal papilloma on contrast-enhanced water-specific, high spatial-resolution and rapid dynamic breast MRI, a retrospective review of unilateral breast images of 15 pathologically proven papilloma was performed. MRI revealed three patterns: Four papillomas were small, smooth, enhancing masses at the posterior end of an enlarged duct, corresponding to the "small lumenal mass" appearance of papilloma known from galactography. MRI detected two of these "small lumenal mass" papillomas in patients with abnormal nipple discharge even when galactography was unsuccessful. Seven papillomas were irregular enhancing masses detected in patients without nipple discharge. None of these papillomas had specifically benign findings. All seven demonstrated rapid enhancement and three showed rim enhancement or spiculation. These "tumor-like" papillomas mimicked invasive breast cancer on MRI. Four papillomas were occult on MRI, not revealed by either contrast-enhanced MRI or fat-suppressed T(2)-weighted MRI. Intraductal papillomas present with a variable appearance on MRI ranging from occult to "small lumenal mass" papillomas to irregular rapidly enhancing lesions that cannot be distinguished from invasive cancers.     

Ergodic stream-lines in steady convection

Stream-lines of steady Rayleigh-Bénard convection with square planform are displayed using Poincaré maps. As the second order mode becomes more important the flow becomes ergodic from the boundaries inward, like a perturbed, integrable hamiltonian system.     

Radical deoxygenation of tertiary alcohols

The thioformates of tertiary alcohols are smoothly reduced by tributyl tin hydride to the corresponding hydrocarbon in a radical chain reaction.     

Familles de graphes expanseurs et paires de HeckeFamilies of expanding graphs and Hecke pairs

Let H be a subgroup of a group G. Suppose that (G,H) is a Hecke pair and that H is finitely generated by a finite symmetric set of size k. Then G/H can be seen as a graph (possibly with loops and multiple edges) whose connected components form a family (X_i)_i∈I of finite k-regular graphs. In this Note, we analyse when the size of these graphs is bounded or tends to infinity and we present criteria for (X_i)_i∈I to be a family of expanding graphs as well as some examples. To cite this article: M.B. Bekka et al., C. R. Acad. Sci. Paris, Ser. I 335 (2002) 463–468.     

The synthesis of organo-imido complexes of Cr(IV) and Fe(III)

The reaction of p-tolylazide with (5,10,15,20- tetraphenylporphyrinato) chromium(II) (Cr(TPP)) yields the high spin chromium(IV) organo-imido complex, CH₃C₆H₄N=Cr(TPP). N,N′-ethylene- bis-(salicylideneiminato)iron(II), (Fe(salen)), however reacts with arylazides to produce iron(III) organo-imido-bridged compounds of general formula, [Fe(salen)]₂NR showing magnetic coupling between the Fe(III) centres.     

Ab initio study of the ground and excited states of HCP and its isomer HPC

The potential energy surface of HCP converting to HPC in its ground electronic state has been investigated with ab initio methods at levels up to MP2/6-311G**//MP4/6-311G** as well as TZV + + ** CASSCF. All geometries are fully optimized and compare favorably to previous theoretical and experimental values. The HCP molecule is predicted to be 85.4 kcal/mol lower in energy than its linear isomer at the-MP2/6-31G*//MP2/6-31G* level. The energy barrier for hydrogen rearrangement is found to be merely 2.3 kcal from the HPC end. CASSCF studies were initiated to clarify the low barrier and lent support to a flat surface as HPC converts to stable, linear HCP at the TZV + + ** level. CASSCF also predicts that HPC is unstable with respect to bending. Harmonic vibrational frequencies for HCP results in 5% accuracy or better. A bent triplet is found to be the lowest excited state using the CASSCF method. © 1993 John Wiley & Sons, Inc.     

Exploitation of reversed micelles as membrane mimetic reagents

The aqueous micelle model and its relation to the various models proposed for aggregation of surfactants in nonaqueous solvents, and the controversy surrounding the applicability of the critical micelle concentration to these aggregates, have been discussed. The effect of the addition of water to, and the nature of acid-base interactions in, surfactants solutions in apolar solvents have been included in a review of the kinetics of the reactivity of esters and other organic sustrates, inoragatic reaction mechanisms, catalysis by solubilized enzymes and electron and proton transfer reactions in these media. Where possible, analogies have been made with membrance mediated processes.     

Self‐Reported Changes in Sun‐Protection Behaviors at Different Latitudes in Australia

Sun exposure is the most important source of vitamin D, but is also a risk factor for skin cancer. This study investigated attitudes toward vitamin D, and changes in sun‐exposure behavior due to concern about adequate vitamin D. Participants (n = 1002) were recruited from four regions of Australia and completed self‐ and interviewer‐administered surveys. Chi‐square tests were used to assess associations between participants' latitude of residence, vitamin D‐related attitudes and changes in sun‐exposure behaviors during the last summer. Multivariate logistic regression analyses were used to model the association between attitudes and behaviors. Overall, people who worried about their vitamin D status were more likely to have altered sun protection and spent more time in the sun people not concerned about vitamin D. Concern about vitamin D was also more common with increasing latitude. Use of novel item response theory analysis highlighted the potential impact of self‐reported behavior change on skin cancer predisposition due concern to vitamin. This cross‐sectional study shows that the strongest determinants of self‐reported sun‐protection behavior changes due to concerns about vitamin D were attitudes and location, with people at higher latitudes worrying more.