Eliminating exposure to aqueous solvents is necessary for the early detection and ultrastructural elemental analysis of sites of calcium and phosphorus enrichment in mineralizing UMR106-01 osteoblastic cultures

The mechanism underlying the mineralization of bone is well studied and yet it remains controversial. Inherent difficulties of imaging mineralized tissues and the aqueous solubility of calcium and phosphate, the 2 ions which combine to form bone mineral crystals, limit current analyses of labile diffusible, amorphous, and crystalline intermediates by electron microscopy. To improve the retention of calcium and phosphorus, we developed a pseudo nonaqueous processing approach and used it to characterize biomineralization foci, extracellular sites of hydroxyapatite deposition in osteoblastic cell cultures. Since mineralization of UMR106-01 osteoblasts is temporally synchronized and begins 78 h after plating, we used these cultures to evaluate the effectiveness of our method when applied to cells just prior to the formation of the first mineral crystals. Our approach combines for the first time 3 well-established methods with a fourth one, i.e. dry ultrathin sectioning. Dry ultrathin sectioning with an oscillating diamond knife was used to produce electron spectroscopic images of mineralized biomineralization foci which were high-pressure frozen and freeze substituted. For comparison, cultures were also treated with conventional processing and wet sectioning. The results show that only the use of pseudo nonaqueous processing was able to detect extracellular sites of early calcium and phosphorus enrichment at 76 h, several hours prior to detection of mineral crystals within biomineralization foci.

Coping personality type and environmental enrichment affect aggression at weaning in pigs

This study investigated the effects of different environmental treatments and personality types on aggression at mixing of newly weaned domestic piglets. From birth to weaning, 16 litters were housed with their dams in either barren (B) or larger, substrate-enriched (E) environments. At 15 days old, piglets were classified as 'high' (HR) or low resistant' (LR) in a manual restraint test (backtest), which is thought to identify proactive (HR) and reactive (LR) stress coping strategies that may reflect different personality types. At 30 days old, 128 piglets were weaned, relocated and mixed into 32 pens comprising two HR and two LR unfamiliar pigs, balanced for sex and weaning weight. Eight B and eight E groups changed environmental condition whereas the others remained in the same type of environment. Number and duration of fights. fight outcomes and unilateral fighting were scored for 5 h post-mixing and skin lesions were counted before and 5 h, 1 day and 2 days after mixing. On the day following weaning, fighting and also exploratory and oral manipulative behaviours were measured for 6 h. Generalized Linear Mixed Model analyses suggested interactions between pre-weaning environment, post-weaning environment and personality type. Overall, pre-weaning E pigs had longer fights at weaning and mixing (P=0.01) and fought for longer on the next day (P=0.02) than pre-weaning B pigs, and inflicted more skin lesions (P=0.02). Post-weaning enrichment did not affect fighting at mixing but reduced the time spent fighting the next day (P=0.03). Personality had subtle and environment-dependent effects on fighting, and influenced the "structure" rather than the amount of aggressive behaviour. HR pigs, for instance, bullied (i.e. chased surrendering pigs) more often (P=0.009) and their fighting behaviour was less affected by their relative body weight than that of LR pigs. Post-weaning E pigs showed relatively higher levels of exploratory behaviour (P=0.02) and less oral manipulative behaviour (P=0.04) than post-weaning B pigs. In particular, switching from a good quality environment (E) to a worse quality one (B) at weaning decreased exploratory behaviour on the next day, especially for LR pigs, who also tended to fight with and orally manipulate their pen mates more in that condition, and seemed to be more affected by a deterioration of the environment. Overall, pre-weaning enrichment increased aggression after weaning whereas post-weaning enrichment reduced it, and personality type related to some aspects of fighting behaviour. (C) 2011 Elsevier B.V. All rights reserved.

Glacier mass balance reconstruction by sublimation induced enrichment of chemical species on Cerro Tapado (Chilean Andes)

A 36 m long ice core down to bedrock from the Cerro Tapado glacier (5536 m a.s.l, 30°08' S, 69°55' W) was analyzed to reconstruct past climatic conditions for Northern Chile. Because of the marked seasonality in the precipitation (short wet winter and extended dry summer periods) in this region, major snow ablation and related post-depositional processes occur on the glacier surface during summer periods. They include predominantly sublimation and dry deposition. Assuming that, like measured during the field campaign, the enrichment of chloride was always related to sublimation, the chemical record along the ice core may be applied to reconstruct the history of such secondary processes linked to the past climatic conditions over northern Chile. For the time period 1962–1999, a mean annual net accumulation of 316 mm water equivalent (weq) and 327 mm weq loss by sublimation was deduced by this method. This corresponds to an initial total annual accumulation of 539 mm weq. The annual variability of the accumulation and sublimation is related with the Southern Oscillation Index (SOI): higher net-accumulation during El-Niño years and more sublimation during La Niña years. The deepest part of the ice record shows a time discontinuity; with an ice body deposited under different climatic conditions: 290 mm higher precipitation but with reduced seasonal distribution (+470 mm in winter and –180 mm in summer) and –3°C lower mean annual temperature. Unfortunately, its age is unknown. The comparison with regional proxy data however let us conclude that the glacier buildup did most likely occur after the dry mid-Holocene.

Generic detection of coronaviruses and differentiation at the prototype strain level by reverse transcription-PCR and nonfluorescent low-density microarray

A nonfluorescent low-cost, low-density oligonucleotide array was designed for detecting the whole coronavirus genus after reverse transcription (RT)-PCR. The limit of detection was 15.7 copies/reaction. The clinical detection limit in patients with severe acute respiratory syndrome was 100 copies/sample. In 39 children suffering from coronavirus 229E, NL63, OC43, or HKU1, the sensitivity was equal to that of individual real-time RT-PCRs.

Mouse enrichment

The provision of environmental enrichment to mice used in research is a topic that generates often widely divergent opinions as to the value of this husbandry practice. The literature is replete with examples of the positive effects enrichment has on the animals’ welfare. However, many published studies have demonstrated that enrichment can impact research in unforeseen and unpredictable ways, and thus may pose a concern as a confounding variable. Indeed, an analysis of the outcomes of murine enrichment studies suggests that both views are accurate, and thus inclusion of environmental enrichment in the mouse cage should be done with the understanding that while the use of appropriate cage complexities can certainly enhance animal welfare and concomitantly provide a better animal model for scientific purposes, the wrong choice of enrichment may negatively impact the welfare of the animals or the research. This chapter describes our current understanding regarding the effects of several commonly used mouse enrichments.

Flavor-phenomenology of two-Higgs-doublet models with generic Yukawa structure

In this article, we perform an extensive study of flavor observables in a two-Higgs-doublet model with generic Yukawa structure (of type III). This model is interesting not only because it is the decoupling limit of the minimal supersymmetric standard model but also because of its rich flavor phenomenology which also allows for sizable effects not only in flavor-changing neutral-current (FCNC) processes but also in tauonic B decays. We examine the possible effects in flavor physics and constrain the model both from tree-level processes and from loop observables. The free parameters of the model are the heavy Higgs mass, tanβ (the ratio of vacuum expectation values) and the “nonholomorphic” Yukawa couplings ϵfij(f=u,d,ℓ). In our analysis we constrain the elements ϵfij in various ways: In a first step we give order of magnitude constraints on ϵfij from ’t Hooft’s naturalness criterion, finding that all ϵfij must be rather small unless the third generation is involved. In a second step, we constrain the Yukawa structure of the type-III two-Higgs-doublet model from tree-level FCNC processes (Bs,d→μ+μ−, KL→μ+μ−, D¯¯¯0→μ+μ−, ΔF=2 processes, τ−→μ−μ+μ−, τ−→e−μ+μ− and μ−→e−e+e−) and observe that all flavor off-diagonal elements of these couplings, except ϵu32,31 and ϵu23,13, must be very small in order to satisfy the current experimental bounds. In a third step, we consider Higgs mediated loop contributions to FCNC processes [b→s(d)γ, Bs,d mixing, K−K¯¯¯ mixing and μ→eγ] finding that also ϵu13 and ϵu23 must be very small, while the bounds on ϵu31 and ϵu32 are especially weak. Furthermore, considering the constraints from electric dipole moments we obtain constrains on some parameters ϵu,ℓij. Taking into account the constraints from FCNC processes we study the size of possible effects in the tauonic B decays (B→τν, B→Dτν and B→D∗τν) as well as in D(s)→τν, D(s)→μν, K(π)→eν, K(π)→μν and τ→K(π)ν which are all sensitive to tree-level charged Higgs exchange. Interestingly, the unconstrained ϵu32,31 are just the elements which directly enter the branching ratios for B→τν, B→Dτν and B→D∗τν. We show that they can explain the deviations from the SM predictions in these processes without fine-tuning. Furthermore, B→τν, B→Dτν and B→D∗τν can even be explained simultaneously. Finally, we give upper limits on the branching ratios of the lepton flavor-violating neutral B meson decays (Bs,d→μe, Bs,d→τe and Bs,d→τμ) and correlate the radiative lepton decays (τ→μγ, τ→eγ and μ→eγ) to the corresponding neutral current lepton decays (τ−→μ−μ+μ−, τ−→e−μ+μ− and μ−→e−e+e−). A detailed Appendix contains all relevant information for the considered processes for general scalar-fermion-fermion couplings.

Cost-effectiveness of ticagrelor and generic clopidogrel in patients with acute coronary syndrome in Switzerland

QUESTION UNDER STUDY The aim of this study was to evaluate the cost-effectiveness of ticagrelor and generic clopidogrel as add-on therapy to acetylsalicylic acid (ASA) in patients with acute coronary syndrome (ACS), from a Swiss perspective. METHODS Based on the PLATelet inhibition and patient Outcomes (PLATO) trial, one-year mean healthcare costs per patient treated with ticagrelor or generic clopidogrel were analysed from a payer perspective in 2011. A two-part decision-analytic model estimated treatment costs, quality-adjusted life years (QALYs), life years and the cost-effectiveness of ticagrelor and generic clopidogrel in patients with ACS up to a lifetime at a discount of 2.5% per annum. Sensitivity analyses were performed. RESULTS Over a patient's lifetime, treatment with ticagrelor generates an additional 0.1694 QALYs and 0.1999 life years at a cost of CHF 260 compared with generic clopidogrel. This results in an Incremental Cost Effectiveness Ratio (ICER) of CHF 1,536 per QALY and CHF 1,301 per life year gained. Ticagrelor dominated generic clopidogrel over the five-year and one-year periods with treatment generating cost savings of CHF 224 and 372 while gaining 0.0461 and 0.0051 QALYs and moreover 0.0517 and 0.0062 life years, respectively. Univariate sensitivity analyses confirmed the dominant position of ticagrelor in the first five years and probabilistic sensitivity analyses showed a high probability of cost-effectiveness over a lifetime. CONCLUSION During the first five years after ACS, treatment with ticagrelor dominates generic clopidogrel in Switzerland. Over a patient's lifetime, ticagrelor is highly cost-effective compared with generic clopidogrel, proven by ICERs significantly below commonly accepted willingness-to-pay thresholds.