Dual-color fluorescence lifetime correlation spectroscopy to quantify protein-protein interactions in live cell

Dual-color fluorescence correlation spectroscopy is an interesting method to quantify protein interaction in living cells. But, when performing these experiments, one must compensate for a known spectral bleed through artifact that corrupts cross-correlation data. In this article, problems with crosstalk were overcome with an approach based on fluorescence lifetime correlation spectroscopy (FLCS). We show that FLCS applied to dual-color EGFP and mCherry cross-correlation allows the determination of protein-protein interactions in living cells without the need of spectral bleed through calibration. The methodology was validated by using EGFP-mCherry tandem in comparison with coexpressed EGFP and mCherry in live cell. The dual-color FLCS experimental procedure where the different laser intensities do not have to be controlled during experiment is really very helpful to study quantitatively protein interactions in live sample.     

Monitoring microbial population dynamics at low densities

We propose a new and simple method for the measurement of microbial concentrations in highly diluted cultures. This method is based on an analysis of the intensity fluctuations of light scattered by microbial cells under laser illumination. Two possible measurement strategies are identified and compared using simulations and measurements of the concentration of gold nanoparticles. Based on this comparison, we show that the concentration of Escherichia coli and Saccharomyces cerevisiae cultures can be easily measured in situ across a concentration range that spans five orders of magnitude. The lowest measurable concentration is three orders of magnitude (1000×) smaller than in current optical density measurements. We show further that this method can also be used to measure the concentration of fluorescent microbial cells. In practice, this new method is well suited to monitor the dynamics of population growth at early colonization of a liquid culture medium. The dynamic data thus obtained are particularly relevant for microbial ecology studies.     

Nitrate and nitrite reduction of a sulphide-rich environment

A sulphide-rich anaerobic sludge acclimated with a molasses wastewater was used to carry out studies on nitrate and nitrite reductions in continuously stirred batch reactors. It was shown that a COD/N-NO_x ratio as high as 65·6 mg mg⁻¹ did not promote dissimilatory reduction of nitrogen oxides to ammonia. Denitrification was characterized by a probable accumulation of gaseous intermediates, nitric oxide (NO) and nitrous oxide (N₂O), by sulphide consumption with concomitant elemental sulphur production and by an increase of the redox potential. In addition, sulphate reducers were completely inhibited by nitrogenous oxides. Cultures performed without any carbon source proved that denitrifiers were able to use sulphides as electron donors. Furthermore, while a lag phase preceded nitrate denitrification, nitrite was consumed immediately. Chemical reduction of nitrite by ferrous iron (Fe²⁺) was considered to be responsible for this difference. Evidence of such a chemodenitrification has been presented by using a sterilized sludge which kept its ability to reduce nitrite while it lost its capacity to use nitrate. Moreover, this chemical activity was favoured by Fe²⁺ addition. Finally, it has been suggested that during the cultures performed with non-sterilized sludge, a biological reduction of the ferric ions (Fe³⁺) would be coupled to nitrite chemodenitrification and would allow a regeneration of Fe²⁺. © 1998 SCI     

Emerging Roles of 1D Vertical Nanostructures in Orchestrating Immune Cell Functions

Engineered nano–bio cellular interfaces driven by 1D vertical nanostructures (1D-VNS) are set to prompt radical progress in modulating cellular processes at the nanoscale. Here, tuneable cell–VNS interfacial interactions are probed and assessed, highlighting the use of 1D-VNS in immunomodulation, and intracellular delivery into immune cells—both crucial in fundamental and translational biomedical research. With programmable topography and adaptable surface functionalization, 1D-VNS provide unique biophysical and biochemical cues to orchestrate innate and adaptive immunity, both ex vivo and in vivo. The intimate nanoscale cell–VNS interface leads to membrane penetration and cellular deformation, facilitating efficient intracellular delivery of diverse bioactive cargoes into hard-to-transfect immune cells. The unsettled interfacial mechanisms reported to be involved in VNS-mediated intracellular delivery are discussed. By identifying up-to-date progress and fundamental challenges of current 1D-VNS technology in immune-cell manipulation, it is hoped that this report gives timely insights for further advances in developing 1D-VNS as a safe, universal, and highly scalable platform for cell engineering and enrichment in advanced cancer immunotherapy such as chimeric antigen receptor-T therapy.     

Negotiating Dialogue Games

Recently in the field of agent communication, many authors have adopted the view of interaction as a joint activity regulated by means of dialogue games. It is argued in particular that this approach should increase the flexibility of dialogues by allowing a variety of game compositions. In this research note, we present a framework suited to this feature. A preliminary attempt to capture the negotiation phase (which allows agents to agree upon the dialogue game currently regulating their conversation) is discussed.     

The Next Generation of Colloidal Probes: A Universal Approach for Soft and Ultra‐Small Particles

The colloidal probe technique, which is based on the atomic force microscope, revolutionizes direct force measurements in many fields, such as interface science or biomechanics. It allows for the first time to determine interaction forces on the single particle or cell level. However, for many applications, important “blind spots” remain, namely, the possibility to probe interaction potentials for nanoparticles or complex colloids with a soft outer shell. Definitely, these are colloidal systems that are currently of major industrial importance and interest from theory. The here‐presented novel approach allows for overcome the aforementioned limitations. Its applicability has been demonstrated for 300 nm sized carboxylate‐modified latex particles as well as sub‐micron core–shell particles with a soft poly‐N‐isopropylacrylamide hydrogel shell and a rigid silica core. For the latter, which until now cannot be studied by the colloidal probe technique, determined is the temperature dependency of electrosteric and adhesion forces has been determined on the single particle level.     

Mu-calculus path checking

We investigate the path model checking problem for the μ-calculus. Surprisingly, restricting to deterministic structures does not allow for more efficient model checking algorithm, as we prove that it can encode any instance of the standard model checking problem for the μ-calculus.     

Fabrication and electroosmotic flow measurements in micro- and nanofluidic channels

An easy method for fabricating micro- and nanofluidic channels, entirely made of a thermally grown silicon dioxide is presented. The nanochannels are up to 1-mm long and have widths and heights down to 200 nm, whereas the microfluidic channels are 20-μm wide and 4.8-μm high. The nanochannels are created at the interface of two silicon wafers. Their fabrication is based on the expansion of growing silicon dioxide and the corresponding reduction in channel cross-section. The embedded silicon dioxide channels were released and are partially freestanding. The transparent and hydrophilic silicon dioxide channel system could be spontaneously filled with aqueous, fluorescent solution. The electrical resistances of the micro- and nanofluidic channel segments were calculated and the found values were confirmed by current measurements. Electrical field strengths up to 600 V/cm were reached within the nanochannels when applying a potential of only 10 V. Electroosmotic flow (EOF) measurements through micro- and nanofluidic channel systems resulted in electroosmotic mobilities in the same order of those encountered in regular, fused silica capillaries.     

Dynamic mechanical properties of hydroxyapatite-reinforced and porous starch-based degradable biomaterials

It has been shown that blends of starch with a poly(ethylene-vinyl-alcohol) copolymer, EVOH, designated as SEVA-C, present an interesting combination of mechanical, degradation and biocompatible properties, specially when filled with hydroxyapatite (HA). Consequently, they may find a range of applications in the biomaterials field. This work evaluated the influence of HA fillers and of blowing agents (used to produce porous architectures) over the viscoelastic properties of SEVA-C polymers, as seen by dynamic mechanical analysis (DMA), in order to speculate on their performances when withstanding cyclic loading in the body. The composite materials presented a promising performance under dynamic mechanical solicitation conditions. Two relaxations were found being attributed to the starch and EVOH phases. The EVOH relaxation process may be very useful in vivo improving the implants performance under cyclic loading. DMA results also showed that it is possible to produce SEVA-C compact surface/porous core architectures with a mechanical performance similar to that of SEVA-C dense materials. This may allow for the use of these materials as bone replacements or scaffolds that must withstand loads when implanted.     

Théodule Ribot (1839–1916), founder of French psychology: A biographical introduction.

Théodule Ribot taught philosophy before obtaining 2 doctoral degrees from the Sorbonne in 1873: 1 about David Hartley and 1 about hereditary factors in psychology. Ribot wrote books on the topics of 19th-century British psychology (1870), 19th-century German psychology (1879), diseases of memory (1881), diseases of will (1883b), and diseases of personality (1885), among others. He founded the Revue Philosophique (Ribot, 1876b), had a chair created for him at the Collège de France (in 1888), helped organize the first French laboratory of experimental psychology (in 1889), and presided over the First International Congress of Psychology in Paris in 1889 (Ribot, 1889b). (PsycINFO Database Record (c) 2010 APA, all rights reserved)