Spatial structure and composition of polysaccharide-protein complexes from Small Angle Neutron Scattering

We use Small Angle Neutron Scattering (SANS), with an original analysis method, to obtain both the characteristic sizes and the inner composition of lysozyme-pectin complexes depending on the charge density. Lysozyme is a globular protein and pectin a natural anionic semiflexible polysaccharide with a degree of methylation (DM) 0, 43 and 74. For our experimental conditions (buffer ionic strength I = 2.5 10-2 mol/L and pH between 3 and 7), the electrostatic charge of lysozyme is always positive (from 8 to 17 depending on pH). The pectin charge per elementary chain segment is negative and can be varied from almost zero to one through the change of DM and pH. The weight molar ratio of lysozyme on pectin monomers is kept constant. The ratio of negative charge content per volume to positive charge content per volume, -/+, is varied between 10 and 0.007. On a local scale, for all charged pectins, a correlation peak appears at 0.2 {\AA}-1 due to proteins clustering inside the complexes. On a large scale, the complexes appear as formed of spherical globules with a well defined radius of 10 to 50 nm, containing a few thousands proteins. The volume fraction Phi of organic matter within the globules derived from SANS absolute cross-sections is around 0.1. The protein stacking, which occurs inside the globules, is enhanced when pectin is more charged, due to pH or DM. The linear charge density of the pectin determines the size of the globules for pectin chains of comparable molecular weights whether it is controlled by the pH or the DM. The radius of the globules varies between 10 nm and 50 nm. In conclusion the structure is driven by electrostatic interactions and not by hydrophobic interactions. The molecular weight also has a large influence on the structure of the complexes since long chains tend to form larger globules. This maybe one reason why DM and pH are not completely equivalent in our system since DM 0 has a short mass, but this may not be the only one. For very low pectin charge (-/+ = 0.07), globules do not appear and the scattering signals a gel-like structure. We did not observe any beads-on-a-string structure

Foaming properties of protein/pectin electrostatic complexes and foam structure at the nanoscale

The foaming properties, foaming capacity and foam stability, of soluble complexes of pectin and a globular protein, napin, have been investigated with a "Foamscan" apparatus. Complementary, we also used SANS with a recent method consisting in an analogy between the SANS by foams and the neutron reflectivity of films to measure in situ film thickness of foams. The effect of ionic strength, of protein concentration and of charge density of the pectin has been analysed. Whereas the foam stability is improved for samples containing soluble complexes, no effect has been noticed on the foam film thickness, which is almost around 315 {\AA} whatever the samples. These results let us specify the role of each specie in the mixture: free proteins contribute to the foaming capacity, provided the initial free protein content in the bulk is sufficient to allow the foam formation, and soluble complexes slow down the drainage by their presence in the Plateau borders, which finally results in the stabilisation of foams

Diel rhythmicity in amino acid uptake by Prochlorococcus

The marine cyanobacterium Prochlorococcus, the most abundant phototrophic organism on Earth, numerically dominates the phytoplankton in nitrogen (N)-depleted oceanic gyres. Alongside inorganic N sources such as nitrite and ammonium, natural populations of this genus also acquire organic N, specifically amino acids. Here, we investigated using isotopic tracer and flow cytometric cell sorting techniques whether amino acid uptake by Prochlorococcus is subject to a diel rhythmicity, and if so, whether this was linked to a specific cell cycle stage. We observed, in contrast to diurnally similar methionine uptake rates by Synechococcus cells, obvious diurnal rhythms in methionine uptake by Prochlorococcus cells in the tropical Atlantic. These rhythms were confirmed using reproducible cyclostat experiments with a light synchronised axenic Prochlorococcus (PCC9511 strain) culture and 35S-methionine and 3H-leucine tracers. Cells acquired the tracers at lower rates around dawn and higher rates around dusk despite >104 times higher concentration of ammonium in the medium, presumably because amino acids can be directly incorporated into protein. Leucine uptake rates by cells in the S+G2 cell cycle stage were consistently 2.2 times higher than those of cells at the G1 stage. Furthermore, S+G2 cells up-regulated amino acid uptake 3.5 times from dawn to dusk to boost protein synthesis prior to cell division. Because Prochlorococcus populations can account from 13% at midday, and up to 42% at dusk, of total microbial uptake of methionine and probably of other amino acids in N-depleted oceanic waters, this genus exerts diurnally variable, strong competitive pressure on other bacterioplankton populations

Structure and spacing of cellulose microfibrils in woody cell walls of dicots

The structure of cellulose microfibrils in situ in wood from the dicotyledonous (hardwood) species cherry and birch, and the vascular tissue from sunflower stems, was examined by wide-angle X-ray and neutron scattering (WAXS and WANS) and small-angle neutron scattering (SANS). Deuteration of accessible cellulose chains followed by WANS showed that these chains were packed at similar spacings to crystalline cellulose, consistent with their inclusion in the microfibril dimensions and with a location at the surface of the microfibrils. Using the Scherrer equation and correcting for considerable lateral disorder, the microfibril dimensions of cherry, birch and sunflower microfibrils perpendicular to the [200] crystal plane were estimated as 3.0, 3.4 and 3.3 nm respectively. The lateral dimensions in other directions were more difficult to correct for disorder but appeared to be 3 nm or less. However for cherry and sunflower, the microfibril spacing estimated by SANS was about 4 nm and was insensitive to the presence of moisture. If the microfibril width was 3 nm as estimated by WAXS, the SANS spacing suggests that a non-cellulosic polymer segment might in places separate the aggregated cellulose microfibrils

Sentinels to climate change. The need for monitoring at South Africa’s Subantarctic laboratory

The International Society for Burns Injuries (ISBI) has published guidelines for the management of multiple or mass burns casualties, and recommends that 'each country has or should have a disaster planning system that addresses its own particular needs.' The need for a national burns disaster plan integrated with national and provincial disaster planning was discussed at the South African Burns Society Congress in 2009, but there was no real involvement in the disaster planning prior to the 2010 World Cup; the country would have been poorly prepared had there been a burns disaster during the event. This article identifies some of the lessons learnt and strategies derived from major burns disasters and burns disaster planning from other regions. Members of the South African Burns Society are undertaking an audit of burns care in South Africa to investigate the feasibility of a national burns disaster plan. This audit (which is still under way) also aims to identify weaknesses of burns care in South Africa and implement improvements where necessary

Du tâtonnement ethnographique au discernement de sens : enquêtes participatives en boucherie et dans la zone d'attente des mineurs isolés étrangers

Partant d’univers de recherche a priori éloignés – la boucherie et la zone d’attente des mineurs isolés étrangers – cet article repose sur une affirmation commune : pour rendre compte des catégories mobilisées par les enquêtés, il faut se mettre en situation de les percevoir. En ce qu’elle rend observable et descriptible le recours à des catégories in situ, l’ethnographie est apparue comme un moyen d’accéder à ce qui est constitutif des expériences de travail. L’enrôlement dans les actions en cours s’est réalisé selon des conditions d’entrée et des modalités distinctes : stagiaire volontaire et experte involontaire. En examinant les configurations d’enquête, l’article montre comment la modulation de la présence participative peut induire le déplacement de la problématique de recherche

Enhanced binding revisited for a spinless particle in non-relativistic QED

We consider a spinless particle coupled to a quantized Bose field and show that such a system has a ground state for two classes of short-range potentials which are alone too weak to have a zero-energy resonance

Impact du Percussionaire® et de la kinésithérapie respiratoire conventionnelle après chirurgie cardiaque : étude randomisée contrôlée

peer reviewedaudience: researcher, professional, studen