In Vitro and In Vivo Evidence that Thrombospondin-1 (TSP-1) Contributes to Stirring- and Shear-Dependent Activation of Platelet-Derived TGF-β1

Thrombospondin 1 (TSP-1), which is contained in platelet α-granules and released with activation, has been shown to activate latent TGF-β1 in vitro, but its in vivo role is unclear as TSP-1-null (Thbs1−/−) mice have a much less severe phenotype than TGF-β1-null (Tgfb1−/−) mice. We recently demonstrated that stirring and/or shear could activate latent TGF-β1 released from platelets and have now studied these methods of TGF-β1 activation in samples from Thbs1−/− mice, which have higher platelet counts and higher levels of total TGF-β1 in their serum than wild type mice. After either two hours of stirring or shear, Thbs1−/− samples demonstrated less TGF-β1 activation (31% and 54% lower levels of active TGF-β1 in serum and platelet releasates, respectively). TGF-β1 activation in Thbs1−/− mice samples was normalized by adding recombinant human TSP-1 (rhTSP-1). Exposure of platelet releasates to shear for one hour led to near depletion of TSP-1, but this could be prevented by preincubating samples with thiol-reactive agents. Moreover, replenishing rhTSP-1 to human platelet releasates after one hour of stirring enhanced TGF-β1 activation. In vivo TGF-β1 activation in carotid artery thrombi was also partially impaired in Thbs1−/− mice. These data indicate that TSP-1 contributes to shear-dependent TGF-β1 activation, thus providing a potential explanation for the inconsistent in vitro data previously reported as well as for the differences in phenotypes of Thbs1−/− and Tgfb1−/− mice

The use of social network analysis to describe the effect of immune activation on group dynamics in pigs

The immune system can influence social motivation with potentially dire consequences for group-housed production animals, such as pigs. The aim of this study was to test the effect of a controlled immune activation in group-housed pigs, through an injection with lipopolysaccharide (LPS) and an intervention with ketoprofen on centrality parameters at the individual level. In addition, we wanted to test the effect of time relative to the injection on general network parameters in order to get a better understanding of changes in social network structures at the group level. 52 female pigs (11-12 weeks) were allocated to four treatments, comprising two injections: ketoprofen-LPS (KL), ketoprofen-saline (KS), saline-LPS (SL) and saline-saline (SS). Social behaviour with a focus on damaging behaviour was observed continuously in 10 x 15 min bouts between 0800 am and 1700 pm 1 day before (baseline) and two subsequent days after injection. Activity was scan-sampled every 5 min for 6 h after the last injection in the pen. Saliva samples were taken for cortisol analysis at baseline and at 4, 24, 48, 72 h after the injections. A controlled immune activation affected centrality parameters for ear manipulation networks at the individual level. Lipopolysaccharide-injected pigs had a lower in-degree centrality, thus, received less interactions, 2 days after the challenge. Treatment effects on tail manipulation and fighting networks were not observed at the individual level. For networks of manipulation of other body parts, in-degree centrality was positively correlated with cortisol response at 4 h and lying behaviour in the first 6 h after the challenge in LPS-injected pigs. Thus, the stronger the pigs reacted to the LPS, the more interactions they received in the subsequent days. The time in relation to injection affected general network parameters for ear manipulation and fighting networks at the group level. For ear manipulation networks, in -degree centralisation was higher on the days following injection, thus, certain individuals in the pen received more interactions than the rest of the group compared to baseline. For fighting networks, betweenness decreased on the first day after injection compared to baseline, indicating that network connectivity increased after the challenge. Networks of tail manipulation and manipulation of other body parts did not change on the days after injection at the group level. Social network analysis is a method that can potentially provide important insights into the effects of sickness on social behaviour in group-housed pigs. (c) 2021 The Authors. Published by Elsevier B.V. on behalf of The Animal Consortium. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).Peer reviewe

Sick and grumpy: changes in social behaviour after a controlled immune stimulation in group-housed gilts

Poor health is associated with an increased risk of tail biting outbreaks in pigs. We propose that this is because illness changes social dynamics either by changing the behaviour of the sick pig towards its penmates, the behaviour of the healthy penmates towards the sick pig, or both. We tested the effect of immune stimulation (lipopolysaccharide (LPS) injection: O111:B4; 1.5 mu g kg(-1) IV) on social behaviour in gilts housed in triplets in a cross-over experiment. Each pen was subjected to the control treatment (all three pigs injected with saline) and then LPS treatment (one pig injected with LPS, two injected with saline), or vice versa. LPS injected pigs had a shift in social motivation and performed more tail- and ear- directed behaviour than saline pigs two days after injection. They seemed to fit the description of 'sick and grumpy'. This change was seen about 40 h after the signs of acute illness dissipated and was not accompanied by a similar increase in activity. We discuss possible mechanisms for this behavioural change in light of changes in neurotransmitter levels at three days after LPS injection described in a previous experiment.Peer reviewe

The Effect of LPS and Ketoprofen on Cytokines, Brain Monoamines, and Social Behavior in Group-Housed Pigs

Poor health is a risk factor for damaging behaviors, but the mechanisms behind this link are unknown. Injection of pigs with lipopolysaccharide (LPS) can be used to model aspects of poor health. Recent studies have shown that LPS-injected pigs perform more tail- and ear-directed behavior compared to saline-injected pigs and suggest that pro-inflammatory cytokines may play a role in these behaviors. The aims of this study were to test the effect of LPS on the social behavior of pigs and the neurotransmitters and modulators in their brains and to test the effect of a nonsteroidal anti-inflammatory drug on the effects of LPS. Fifty-two female pigs (11-12 weeks) were allocated to four treatments comprising two injections: saline-saline (SS), saline-LPS (SL), ketoprofen-saline (KS), and ketoprofen-LPS (KL). Activity was scan-sampled every 5 min for 6 h after the last injection in the pen. Social behavior was observed continuously in 10 x 15-min bouts between 8 a.m. and 5 p.m. 1 day before (baseline) and 1 and 2 days after the injection. Saliva was analyzed for cortisol and plasma for tryptophan and kynurenine. The frontal cortex, hippocampus, hypothalamus, and brain stem were sampled 72 h after the injection and analyzed for cytokines and monoamines. LPS activated the HPA axis and decreased the activity within 6 h after the injection. Ketoprofen lowered the effect of LPS on cortisol release and attenuated the behavioral signs of sickness in challenged pigs. SL pigs manipulated the ears of their pen mates significantly longer than SS pigs 2 days after the injection. LPS had no observed effect on IFN-gamma, TNF-alpha, and IL-18. At 72 h after the injection, plasma tryptophan was depleted in SL pigs, and tryptophan and kynurenine concentrations in the frontal cortex and brain stem of SL pigs were significantly lower compared to those in SS pigs. Dopamine concentrations in the hypothalamus of SL pigs were significantly lower compared to those in SS pigs. Serotonin concentrations in the hypothalamus and noradrenaline concentrations in the hippocampus of SL pigs were significantly lower compared to those in KL pigs. In conclusion, LPS influenced the different neurotransmitters and modulators in the brain that are hypothesized to play an important role in the regulation of mood and behavior.Peer reviewe

Measurements, Models, Systems and Design

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A Proposed Role for Pro-Inflammatory Cytokines in Damaging Behavior in Pigs

Sickness can change our mood for the worse, leaving us sad, lethargic, grumpy and less socially inclined. This mood change is part of a set of behavioral symptoms called sickness behavior and has features in common with core symptoms of depression. Therefore, the physiological changes induced by immune activation, for example following infection, are in the spotlight for explaining mechanisms behind mental health challenges such as depression. While humans may take a day off and isolate themselves until they feel better, farm animals housed in groups have only limited possibilities for social withdrawal. We suggest that immune activation could be a major factor influencing social interactions in pigs, with outbreaks of damaging behavior such as tail biting as a possible result. The hypothesis presented here is that the effects of several known risk factors for tail biting are mediated by pro-inflammatory cytokines, proteins produced by the immune system, and their effect on neurotransmitter systems. We describe the background for and implications of this hypothesis.Peer reviewe

MOA-2009-BLG-387Lb: A massive planet orbiting an M dwarf

We report the discovery of a planet with a high planet-to-star mass ratio in the microlensing event MOA-2009-BLG-387, which exhibited pronounced deviations over a 12-day interval, one of the longest for any planetary event. The host is an M dwarf, with a mass in the range 0.07 M_sun < M_host < 0.49M_sun at 90% confidence. The planet-star mass ratio q = 0.0132 +- 0.003 has been measured extremely well, so at the best-estimated host mass, the planet mass is m_p = 2.6 Jupiter masses for the median host mass, M = 0.19 M_sun. The host mass is determined from two "higher order" microlensing parameters. One of these, the angular Einstein radius \theta_E = 0.31 +- 0.03 mas, is very well measured, but the other (the microlens parallax \pi_E, which is due to the Earth's orbital motion) is highly degenate with the orbital motion of the planet. We statistically resolve the degeneracy between Earth and planet orbital effects by imposing priors from a Galactic model that specifies the positions and velocities of lenses and sources and a Kepler model of orbits. The 90% confidence intervals for the distance, semi-major axis, and period of the planet are 3.5 kpc < D_L < 7.9 kpc, 1.1 AU < a < 2.7AU, and 3.8 yr < P < 7.6 yr, respectively.Comment: 20 pages including 8 figures. A&A 529 102 (2011

Frequency of Solar-Like Systems and of Ice and Gas Giants Beyond the Snow Line from High-Magnification Microlensing Events in 2005-2008

We present the first measurement of planet frequency beyond the "snow line" for planet/star mass-ratios[-4.5<log q<-2]: d^2 N/dlog q/dlog s=(0.36+-0.15)/dex^2 at mean mass ratio q=5e-4, and consistent with being flat in log projected separation, s. Our result is based on a sample of 6 planets detected from intensive follow-up of high-mag (A>200) microlensing events during 2005-8. The sample host stars have typical mass M_host 0.5 Msun, and detection is sensitive to planets over a range of projected separations (R_E/s_max,R_E*s_max), where R_E 3.5 AU sqrt(M_host/Msun) is the Einstein radius and s_max (q/5e-5)^{2/3}, corresponding to deprojected separations ~3 times the "snow line". Though frenetic, the observations constitute a "controlled experiment", which permits measurement of absolute planet frequency. High-mag events are rare, but the high-mag channel is efficient: half of high-mag events were successfully monitored and half of these yielded planet detections. The planet frequency derived from microlensing is a factor 7 larger than from RV studies at factor ~25 smaller separations [2<P<2000 days]. However, this difference is basically consistent with the gradient derived from RV studies (when extrapolated well beyond the separations from which it is measured). This suggests a universal separation distribution across 2 dex in semi-major axis, 2 dex in mass ratio, and 0.3 dex in host mass. Finally, if all planetary systems were "analogs" of the Solar System, our sample would have yielded 18.2 planets (11.4 "Jupiters", 6.4 "Saturns", 0.3 "Uranuses", 0.2 "Neptunes") including 6.1 systems with 2 or more planet detections. This compares to 6 planets including one 2-planet system in the actual sample, implying a first estimate of 1/6 for the frequency of solar-like systems.Comment: 42 pages, 10 figure

Extreme Magnification Microlensing Event OGLE-2008-BLG-279: Strong Limits on Planetary Companions to the Lens Star

We analyze the extreme high-magnification microlensing event OGLE-2008-BLG-279, which peaked at a maximum magnification of A ~ 1600 on 30 May 2008. The peak of this event exhibits both finite-source effects and terrestrial parallax, from which we determine the mass of the lens, M_l=0.64 +/- 0.10 M_Sun, and its distance, D_l = 4.0 +/- 0.6. We rule out Jupiter-mass planetary companions to the lens star for projected separations in the range 0.5-20 AU. More generally, we find that this event was sensitive to planets with masses as small as 0.2 M_Earth ~= 2 M_Mars with projected separations near the Einstein ring (~3 AU).Comment: 25 pages, 7 figures, submitted to Ap

Microlensing Event MOA-2007-BLG-400: Exhuming the Buried Signature of a Cool, Jovian-Mass Planet

We report the detection of the cool, Jovian-mass planet MOA-2007-BLG-400Lb. The planet was detected in a high-magnification microlensing event (with peak magnification A_max = 628) in which the primary lens transited the source, resulting in a dramatic smoothing of the peak of the event. The angular extent of the region of perturbation due to the planet is significantly smaller than the angular size of the source, and as a result the planetary signature is also smoothed out by the finite source size. Thus the deviation from a single-lens fit is broad and relatively weak (~ few percent). Nevertheless, we demonstrate that the planetary nature of the deviation can be unambiguously ascertained from the gross features of the residuals, and detailed analysis yields a fairly precise planet/star mass ratio of q = 0.0026+/-0.0004, in accord with the large significance (\Delta\chi^2=1070) of the detection. The planet/star projected separation is subject to a strong close/wide degeneracy, leading to two indistinguishable solutions that differ in separation by a factor of ~8.5. Upper limits on flux from the lens constrain its mass to be M < 0.75 M_Sun (assuming it is a main-sequence star). A Bayesian analysis that includes all available observational constraints indicates a primary in the Galactic bulge with a mass of ~0.2-0.5 M_Sun and thus a planet mass of ~ 0.5-1.3 M_Jupiter. The separation and equilibrium temperature are ~0.6-1.1AU (~5.3-9.7AU) and ~103K (~34K) for the close (wide) solution. If the primary is a main-sequence star, follow-up observations would enable the detection of its light and so a measurement of its mass and distance.Comment: 30 pages, 6 figures, Submitted to Ap