Impact of the virus purification protocol on aggregation and electrokinetics of MS2 phages and corresponding virus-like particles.

Previous experimental and theoretical studies have established that electrokinetic and aggregation properties of soft MS2 phages are not only governed by the physico-chemical features of their proteinaceous outer surface but are also significantly impacted by those of their inner RNA component (Dika et al. Appl. Environ. Microbiol., 2011, 14, 4939-4948). These conclusions contradict the recent findings of Nguyen et al. (Soft Matter, 2011, 7, 10449-10456) who reported identical electrokinetic and aggregation characteristics for MS2 and corresponding virus like particles (VLPs) that lack the internal RNA component. We demonstrate here that this contradiction originates from the different purification methods adopted prior to measurements. More generally, we show that stability and electrohydrodynamics of viruses differ according to purification by (i) dialysis, (ii) isopycnic centrifugation in the cesium chloride gradient, and (iii) precipitation using polyethylene glycol (PEG). Methods (i) and (iii) lead to aggregation of MS2 phages at pH ≤ 4 and pH ≤ 6 in 1-100 mM NaNO3 solutions, respectively, while under such conditions aggregation is not observed for MS2 and VLP suspensions prepared according to (ii). In addition, VLPs prepared following methods (i) and (iii) aggregate only at the isoelectric point (pH ~ 3-4) in 1 mM NaNO3 solution. Electrophoretic mobility data of stable MS2 and VLP particles were further examined using a recent formalism for electrokinetics of soft multilayered colloids. The analysis qualitatively shows how the purification protocol may affect either the outer particle surface properties and/or the inner particle content. Finally, the non-DLVO aggregation behavior of MS2 and VLPs purified via the above protocols is discussed in terms of the possible change in corresponding interparticular interactions.

[Health Care Insurance in France: its impact on income distribution between age and social groups]. / Redistribution des revenus opérée par l'assurance maladie entre groupes d'âge et groupes sociaux.

Our study, based on microsimulation models, evaluates the redistributive impact of health care insurance in France on income distribution between age and social groups. This work sheds light on the debate concerning the respective role of the public health care insurance (PHI) and the private supplemental health care insurance (SHI) in France. The analysis points out that the PHI enables the lowest-income households and the pensioners a better access to health care than they would have had under a complete private SHI. Due to the progressivity of taxes, low-income households contribute less to the PHI and get higher benefits because of a weaker health. Pensioners have low contributions to public health care finance but the highest health care expenditures.

Surface properties of bacteria sensitive and resistant to the class IIa carnobacteriocin Cbn BM1.

AIMS: Class IIa bacteriocins are small antimicrobial peptides synthesized by lactic acid bacteria. The proposed mechanisms of action for class IIa bacteriocins suggest that the physicochemical properties of the target bacterial surface govern the bacteriocin antimicrobial activity. The aim of this study is to decipher the relationship between both sensitivity and resistance to a class IIa bacteriocin, carnobacteriocin BM1 and physicochemical surface properties of bacteria. METHODS AND RESULTS: The study was performed on 18 strains by a microbial adhesion to solvents process and with electrophoretic mobility measurements considering bacteria as soft particles. A large variation in bacterial surface properties is observed among the bacterial populations. Electro-hydrodynamic parameters values appear to be more homogeneous for sensitive strains than for the resistant ones, which can exhibit more extreme values. CONCLUSIONS: Physicochemical surface properties of 18 strains determined show large variations between the strains. However, no direct link between these surface properties and the resistant/sensitive phenotypes of the strains can be stated. SIGNIFICANCE AND IMPACT OF THE STUDY: The surface physicochemical properties tested have a low predictive power to discriminate sensitive or resistant strains when determined at the bacterial population scale.

Carbon contamination of soft X-ray beamlines: dramatic anti-reflection coating effects observed in the 1 keV photon energy region.

Carbon contamination is a general problem of under-vacuum optics submitted to high fluence. In soft X-ray beamlines carbon deposit on optics is known to absorb and scatter radiation close to the C K-edge (280 eV), forbidding effective measurements in this spectral region. Here the observation of strong reflectivity losses is reported related to carbon deposition at much higher energies around 1000 eV, where carbon absorptivity is small. It is shown that the observed effect can be modelled as a destructive interference from a homogeneous carbon thin film.

Impact of internal RNA on aggregation and electrokinetics of viruses: comparison between MS2 phage and corresponding virus-like particles.

We compare for the first time the electrokinetic and aggregation properties of MS2 phage (pH 2.5 to 7, 1 to 100 mM NaNO(3) electrolyte concentration) with those of the corresponding virus-like particles (VLPs), which lack entirely the inner viral RNA component. In line with our previous work (J. Langlet, F. Gaboriaud, C. Gantzer, and J. F. L. Duval, Biophys. J. 94:3293-3312, 2008), it is found that modifying the content of RNA within the virus leads to very distinct electrohydrodynamic and aggregation profiles for MS2 and MS2 VLPs. Under the given pH and concentration conditions, MS2 VLPs exhibit electrophoretic mobility larger in magnitude than that of MS2, and both have similar isoelectric point (IEP) values (∼4). The electrokinetic results reflect a greater permeability of MS2 VLPs to electroosmotic flow, developed within/around these soft particles during their migration under the action of the applied electrical field. Results also support the presence of some remaining negatively charged component within the VLPs. In addition, MS2 phage systematically forms aggregates at pH values below the IEP, regardless of the magnitude of the solution ionic strength, whereas MS2 VLPs aggregate under the strict condition where the pH is relatively equal to the IEP at sufficiently low salt concentrations (<10 mM). It is argued that the stability of VLPs against aggregation and the differences between electrokinetics of MS2 and corresponding VLPs conform to recently developed formalisms for the stability and electrohydrodynamics of soft multilayered particles. The differences between the surface properties of these two kinds of particles reported here suggest that VLPs may not be appropriate for predicting the behavior of pathogenic viruses in aqueous media.

Effects of a participatory approach, with systematic impact matrix analysis in herd health planning in organic dairy cattle herds.

The animal health and welfare status in European organic dairy production does not in all aspects meet the organic principles and consumers' expectations and needs to be improved. To achieve this, tailored herd health planning, targeted to the specific situation of individual farms could be of use. The aim of this study was to apply herd health planning in a structured participatory approach, with impact matrix analysis, not previously used in this context, in European organic dairy farms and to assess changes in animal health and welfare. Herd health planning farm visits were conducted on 122 organic dairy farms in France, Germany and Sweden. The farmer, the herd veterinarian and/or an advisor took part in the farm discussions. The researcher served as facilitator. Baseline data on the animal health status of the individual farm, collected from national milk recording schemes, were presented as an input for the discussion. Thereafter a systematic impact matrix analysis was performed. This was to capture the complexity of individual farms with the aim to identify the farm-specific factors that could have a strong impact on animal health. The participants (i.e. farmer, veterinarian and advisor) jointly identified areas in need of improvement, taking the health status and the interconnected farm system components into account, and appropriate actions were jointly identified. The researcher took minutes during the discussions, and these were shared with the participants. No intervention was made by the researcher, and further actions were left with the participants. The number of actions per farm ranged from 0 to 22. The change in mortality, metabolic diseases, reproductive performance and udder health was assessed at two time points, and potential determinators of the change were evaluated with linear regression models. A significant association was seen between change in udder health, as measured by the somatic cell count, and country. At the first follow-up, a significant association was also found between change in the proportion of prolonged calving interval and the farmers' desire to improve reproductive health as well as with an increase in herd size, but this was not seen at the second follow-up. The degree of implementation of the actions was good (median 67%, lower quartile 40%, upper quartile 83%). To conclude, the degree of implementation was quite high, improvement of animal health could not be linked to the herd health planning approach. However, the approach was highly appreciated by the participants and deserves further study.

Farm-specific failure costs of production disorders in European organic dairy herds.

On-farm decision support in animal health management requires a tailor-made failure costs (FCs) assessment of production disorders for the individual farm. In our study we defined a generic framework to estimate the FC of production disorders in dairy cows. We converted the framework to a practical tool in which the farm-specific FC of mastitis, ketosis, lameness and metritis were estimated for 162 organic dairy farms in four European countries. Along with the structure of the framework, the FC estimation required three distinct types of model input: performance input (related to herd performance parameters), consequential input (related to the consequences of the disorders) and economic input (related to price levels). Input was derived from official herd recordings (e.g. test-day records and animal health recordings) and farmers' responses (e.g. questionnaire replies). The average FC of mastitis, ketosis, lameness and metritis amounted to € 96, € 21, € 43 and € 10 per cow per year, respectively. The variation in FC outcomes was high among farmers and countries. Overall ranking of the disorders based on absolute values was the same for all countries, with mastitis being the costliest disorder followed in order by lameness, ketosis, and metritis. Farm specific estimates can be used to rank production related disorders in terms of their associated failure costs and thus provide valuable insights for herd health management. The practical calculation tool developed in this study should be considered by farmers or herd health advisors to support their animal health practices or advice.

Tunable High Spatio-Spectral Purity Undulator Radiation from a Transported Laser Plasma Accelerated Electron Beam.

Undulator based synchrotron light sources and Free Electron Lasers (FELs) are valuable modern probes of matter with high temporal and spatial resolution. Laser Plasma Accelerators (LPAs), delivering GeV electron beams in few centimeters, are good candidates for future compact light sources. However the barriers set by the large energy spread, divergence and shot-to-shot fluctuations require a specific transport line, to shape the electron beam phase space for achieving ultrashort undulator synchrotron radiation suitable for users and even for achieving FEL amplification. Proof-of-principle LPA based undulator emission, with strong electron focusing or transport, does not yet exhibit the full specific radiation properties. We report on the generation of undulator radiation with an LPA beam based manipulation in a dedicated transport line with versatile properties. After evidencing the specific spatio-spectral signature, we tune the resonant wavelength within 200-300 nm by modification of the electron beam energy and the undulator field. We achieve a wavelength stability of 2.6%. We demonstrate that we can control the spatio-spectral purity and spectral brightness by reducing the energy range inside the chicane. We have also observed the second harmonic emission of the undulator.

Evaluation of the impact of a Herd Health and Production Management programme in organic dairy cattle farms: a process evaluation approach.

Animal health planning activities are not always providing a satisfactory positive impact on herd health and welfare. Moreover, evaluating the impact of advisory programmes is complex due to multiple interacting elements that influence its outcome. Therefore, measuring solely health outcomes is not sufficient: the whole process of the implementation and use of such programmes should be evaluated. In order to evaluate the impact of an intervention with a Herd Health and Production Management (HHPM) programme a process evaluation framework was designed and used. The intervention involved 20 organic dairy cattle farmers and their advisors, in both France and Sweden. In both countries 20 organic dairy farms were selected as control herds. The evaluation of the HHPM programme was based on: (a) the compliance to the programme; (b) the programme's functions influencing herd health management practices and stimulating dialogue between farmers and advisors; (c) its effectiveness in terms of improving herd health compared with control farms. Complete compliance to the programme was fulfilled by 21 out of 40 farmers-advisors. Results from a questionnaire showed that the programme functioned as intended (e.g. by allowing early identification of herd health problems), stimulated change in farmers' herd health management practices and farmer-advisor dialogue. Even though the majority of the users perceived that the programme contributed to herd health improvements, no significant differences in health outcomes were found when compared with control farms 12 months after the start of the intervention. The programme allowed creating an environment promoting the exchange of information between farmers and advisors, necessary to define pertinent advice in a farm-specific situation. Future research should aim at improving methods for the evaluation of the effect of advisory programmes, by identifying early indicators for effective advice and developing methods to evaluate the quality of advisory situations without interfering with them.

Control of laser plasma accelerated electrons for light sources.

With gigaelectron-volts per centimetre energy gains and femtosecond electron beams, laser wakefield acceleration (LWFA) is a promising candidate for applications, such as ultrafast electron diffraction, multistaged colliders and radiation sources (betatron, compton, undulator, free electron laser). However, for some of these applications, the beam performance, for example, energy spread, divergence and shot-to-shot fluctuations, need a drastic improvement. Here, we show that, using a dedicated transport line, we can mitigate these initial weaknesses. We demonstrate that we can manipulate the beam longitudinal and transverse phase-space of the presently available LWFA beams. Indeed, we separately correct orbit mis-steerings and minimise dispersion thanks to specially designed variable strength quadrupoles, and select the useful energy range passing through a slit in a magnetic chicane. Therefore, this matched electron beam leads to the successful observation of undulator synchrotron radiation after an 8 m transport path. These results pave the way to applications demanding in terms of beam quality.

Publisher Correction: Control of laser plasma accelerated electrons for light sources.

The original version of this Article contained an error in the last sentence of the first paragraph of the Introduction and incorrectly read 'A proper electron beam control is one of the main challenges towards the Graal of developing a compact alternative of X-ray free-electron lasers by coupling LWFA gigaelectron-volts per centimetre acceleration gradient with undulators in the amplification regime in equation 11, nx(n-ß) x ß: n the two times and beta the two times should be bold since they are vectorsin Eq. 12, ß should be bold as well.' The correct version is 'A proper electron beam control is one of the main challenges towards the Graal of developing a compact alternative of X-ray free-electron lasers by coupling LWFA gigaelectron-volts per centimetre acceleration gradient with undulators in the amplification regime.'This has been corrected in both the PDF and HTML versions of the Article.

How can veterinarians be interesting partners for organic dairy farmers? French farmers' point of views.

Organic dairy farmers must live up to the organic goal of 'good health' in respect the organic principles and regulation. Veterinarians could be the organic dairy farmers' expected sparring partners in reaching this goal but have found difficulties to establish advisory relationships with them. The objectives of this study are -from organic dairy farmers' points of view- (i) to describe farmers' objectives and strategies regarding herd health, (ii) to describe private veterinarians' roles in farmers' animal health promotion strategies and (iii) to identify farmers' reasons for accepting veterinarians in an advisory role. Fourteen organic dairy farmers were interviewed using qualitative research interviews. Data collection and analysis was performed using a modified approach to Grounded Theory. Organic dairy farmers had animal health management strategies focusing on animal health promotion. Veterinarians had most often solely the role of therapist in farmers' animal health management strategies. Reasons explaining that veterinarians were not able to establish advisory roles were found in the differences between veterinarians and farmers regarding their animal health strategies and solutions to disease problems. Furthermore, veterinarians did not always share farmers' (organic) objectives, values and priorities and this could lead to disagreement on the best choice in animal health management practices. This might be further amplified in situations where there exists a lack of dialogue and mutual interest in other.

The Tragic Tale of a Father and Son: an Unusual Patricide.

Parricide is a category of homicide in which the victims are the parents, and the killers, their children. The authors report a case of a 45-year-old man who killed his 73-year-old widowed father in an extremely violent manner: he struck the father in the head with a wooden wash board, stabbed him with a pair of scissors and several times with a screwdriver. Afterwards, he kicked the victim in the face and jumped up and down on his body, thereby crushing the father's chest with his weight. The case reported here may be classified as an "unusual patricide" when one considers the age of the aggressor, the diagnosis of psychiatric disorders, that the victim was the father, and that the murder was committed in an extremely violent manner.

Geochemistry of bed and suspended sediment in the Mississippi river system: provenance versus weathering and winnowing.

Stream-bed sediment for the size fraction less than 150 microm, examined in 14,000 samples collected mostly from minor tributaries to the major rivers throughout the Mississippi River drainage system, is composed of 5 mineral fractions identified by factor analysis-Al-silicate minerals, quartz, calcite and dolomite, heavy minerals, and an Fe-Mn fraction. The Al-silicate fraction parallels its distribution in the regolith, emphasizing the local sediment source as a primary control to its distribution. Quartz and the heavy-mineral fraction, and associated trace elements, exhibit a complementary distribution to that of the Al-silicate fraction, with a level of enrichment in the bed sediment that is achieved through winnowing and sorting. The carbonate fraction has a distribution suggesting its dissolution during transport. Trace elements partitioned onto the Fe-Mn, possibly amorphous oxyhydride, fraction are introduced to the streams, in part, through human activity. Except for the heavy-mineral fraction, these fractions are identified in suspended sediment from the Mississippi River itself. Although comparison of the tributary bed sediment with the riverine suspended sediment is problematic, the geochemistry of the suspended sediment seems to corroborate the interpretation of the geochemistry of the bed sediment.

Perceptions of French private veterinary practitioners' on their role in organic dairy farms and opportunities to improve their advisory services for organic dairy farmers.

Veterinarians could be the expected sparring partners of organic dairy farmers in promoting animal health which is one of the main organic principles. However, in the past organic dairy farmers did not always consider veterinarians to be pertinent advisors for them. The objectives of this study are - from private veterinary practitioners' point of views- i) to describe the roles of veterinarians today in organic dairy farmers' animal health promotion strategies, ii) to identify factors related to organic farming which determine their role on organic dairy farms, and, iii) to identify opportunities for improvement of veterinarians' advisory services for organic dairy herds. Fourteen veterinarians, providing herd health advisory services to dairy farmers, were interviewed using qualitative semi-structured research interviews. A modified approach to Grounded Theory was used for data collection and analysis. Most often veterinarians had only contact with the organic dairy farmers in cases of individual ill animals or acute herd health problems. Even though certain veterinarians experienced situations and approaches of animal health and welfare on organic dairy farms not meeting their standards, they were not always able to establish themselves an advisory role supporting farmers in improving this. Indeed, organic production principles, regulations and farmers' health approaches challenged veterinarians' values on animal health and welfare and their perceptions of 'good veterinary practices'. Also, some veterinarians considered that there was no direct economic interest for them in the organic dairy sector and that could diminish their willingness to invest in this sector. Possible opportunities for improvement were identified; for example proposing more proactively advice via existing organisations, by making adaptations to advisory services for the organic sector and/or by dissociating veterinarians' curative role from their advisory role in disease prevention.

A participatory approach to design monitoring indicators of production diseases in organic dairy farms.

Production diseases have an important negative effect on the health and welfare of dairy cows. Although organic animal production systems aim for high animal health levels, compliance with European organic farming regulations does not guarantee that this is achieved. Herd health and production management (HHPM) programs aim at optimizing herd health by preventing disease and production problems, but as yet they have not been consistently implemented by farmers. We hypothesize that one reason is the mismatch between what scientists propose as indicators for herd health monitoring and what farmers would like to use. Herd health monitoring is a key element in HHPM programs as it permits a regular assessment of the functioning of the different components of the production process. Planned observations or measurements of these components are indispensable for this monitoring. In this study, a participatory approach was used to create an environment in which farmers could adapt the indicators proposed by scientists for monitoring the five main production diseases on dairy cattle farms. The adaptations of the indicators were characterized and the farmers' explanations for the changes made were described. The study was conducted in France and Sweden, which differ in terms of their national organic regulations and existing advisory services. In both countries, twenty certified organic dairy farmers and their animal health management advisors participated in the study. All of the farmers adapted the initial monitoring plan proposed by scientists to specific production and animal health situation on their farm. This resulted in forty unique and farm-specific combinations of indicators for herd health monitoring. All but three farmers intended to monitor five health topics simultaneously using the constructed indicators. The qualitative analysis of the explanations given by farmers for their choices enabled an understanding of farmers' reasons for selecting and adapting indicators. This is valuable information for scientists involved in the design of HHPM programs. Advisors in the field also can benefit from this participatory approach because it transforms monitoring tools provided by scientists into farm-specific tools.

Energy transfers from photosystem II to photosystem I in Cryptomonas rufescens (Cryptophyceae).

In Cryptomonas rufescens (Cryptophyceae), phycoerythrin located in the thylakoid lumen is the major accessory pigment. Oxygen action spectra prove phycoerythrin to be efficient in trapping light energy. The fluorescence excitation spectra at -196 degrees C obtained by the method of Butler and Kitajima (Butler, W.L. and Kitajima, M. (1975) Biochim. Biophys. Acta 396, 72-85) indicate that like in Rhodophyceae, chlorophyll a is the exclusive light-harvesting pigment for Photosystem I. For Photosystem II we can observe two types of antennae: (1) a light-harvesting chlorophyll complex connected to Photosystem II reaction centers, which transfers excitation energy to Photosystem I reaction centers when all the Photosystem II traps are closed. (2) A light-harvesting phycoerythrin complex, which transfers excitation energy exclusively to the Photosystem II reaction complexes responsible for fluorescence at 690 nm. We conclude that in Cryptophycease, phycoerythrin is an efficient light-harvesting pigment, organized as an antenna connected to Photosystem II centers, antenna situated in the lumen of the thylakoid. However, we cannot afford to exclude that a few parts of phycobilin pigments could be connected to inactive chlorophylls fluorescing at 690 nm.

Differential expression of secretogranin II and chromogranin A genes in the female rat pituitary through sexual maturation and estrous cycle.

Secretogranin II (SgII) is a protein of pituitary secretory granules released by LHRH-stimulated gonadotrope cells. Estrogens and androgens are modulators of SgII release. Experiments were performed to determine the regulation of expression of the SgII gene in the female rat pituitary, during sexual maturation and according to the estrous cycle. Age- and cycle-related changes in SgII mRNA content were estimated through cytoplasmic slot blot; SgII content was determined by western blotting; maturation of the protein was controlled through [35S]sulfate labeling. Variations in chromogranin A (CgA), another protein of secretory granules, were analyzed in the same experimental conditions to assess the specificity of SgII regulation. The pituitary SgII concentration increased between days 7 and 21 (2.2-fold) and then declined to the initial 7-day-old value. Simultaneously, the CgA concentration went through a maximum between days 14 and 21 and then strongly dropped to barely detectable levels in the adult pituitary. The SgII mRNA concentration followed roughly the same pattern as the protein. Moreover, the sulfation level remained constant between days 14 and 60. These results demonstrated a regulatory mechanism operating, during sexual maturation, on the SgII gene and not on the protein processing or on storage/release steps. In the 4-day cycling females, the pituitary SgII mRNA and protein contents were the lowest during estrus. They then increased to their highest values in diestrus II. Moreover, the sulfation level of SgII was significantly higher during estrus than during any other stage. Due to its low content level, variations in pituitary CgA could not be demonstrated during the cycle.

Differential distribution of androgen and estrogen receptors in rat pituitary cell populations separated by centrifugal elutriation.

The distribution of 5 alpha-dihydrotestosterone (DHT)-binding sites within specific pituitary cell populations, as compared with the estradiol (E2)-binding sites distribution, could be estimated after gonadotrope cell enrichment. Centrifugal elutriation allowed the purification of large gonadotropes from 42-day-old male rat pituitaries; this population contained 5.3 +/- 1.0% of the cells, 54 +/- 3% of the LH, and less than 5% of PRL. A gonadotrope-depleted fraction was also defined that contained more than 92% of the total PRL and was thus called the lactotrope population though it still contained small gonadotropes and most somatotropes. DHT and E2 receptors were quantified by multidose saturation analysis. The association constants (Ka) ranged from 0.2-0.5 X 10(9) M-1 both for E2 and DHT independent of the cell population. The binding of DHT was mainly observed in the gonadotropes (50-100 fmol/10(6) cells), while it was very low in the other cells (0.9-3 fmol/10(6) cells). E2-binding sites measured within the same cell populations showed a broader distribution than DHT-binding sites. Whereas the ratio of binding DHT/E2 averaged 0.86 +/- 0.01 in the gonadotropes, it reached only 0.35 +/- 0.07 in the lactotropes. Thus, using enriched cell populations, our results demonstrate for the first time a quantitative selective distribution of androgen-binding sites as compared with the estrogen sites.