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Even during ongoing global biodiversity losses and extinctions, numerous species have shown recoveries in terms of increased abundance and/or range extent. Understanding the mechanisms that contribute to, or limit, these recoveries is critical not just to ensure they continue, but to promote similar recoveries across broader ecosystems. Here, we explore the changes in abundance and range extent of selected 47 recovering species (24 mammals and 23 birds) in Europe using official data reported by EU Member States and supplemented using the Living Planet Index database. We investigate how the diversity of ongoing threats and conservation measures contribute to the likelihood and extent of recoveries. For birds, long-term recoveries were less likely among species impacted by a greater diversity of threats, although this may be mitigated by the diversity of conservation measures in place. Similarly, for mammals, populations with reported threats recovered less quickly while those with management actions in place recovered more quickly. To achieve the aims of the UN Decade on Restoration, we need to ensure, even for recovering species, that threats continue to be reduced and that conservation management actions are ongoing and effective.

Obesity is becoming a critical problem in the developed world and is associated with an increased incidence of osteoarthritis of the hip. The Oxford Hip Score was used to determine if Body Mass Index (BMI) is an independent factor in determining patient outcome following primary total hip arthroplasty (THA). Using data from 353 operations we found that patients with BMI ≥ 30 had an absolute score that was lower preoperatively and postoperatively compared to those with a BMI < 30. There was no difference in pre- and postoperative point score change within each group; Kendall’s rank correlation was 0.00047 (95% CI, −0.073 to 0.074 (p=0.99)) and demonstrated no trend. There was no statistically significant difference in change between those with BMI ≥ 30 and < 30 (p=0.65). We suggest that those with a higher BMI be considered for THA as they can expect the same degree of improvement as those with a lower BMI. Given the on-going increase in obesity these findings could be significant for the future of THA.

Adenosine 5′-monophosphate–activated protein kinase (AMPK) is a pivotal regulator of metabolism at cellular and organismal levels. AMPK also suppresses inflammation. We found that pharmacological activation of AMPK rapidly inhibited the Janus kinase (JAK)–signal transducer and activator of transcription (STAT) pathway in various cells. In vitro kinase assays revealed that AMPK directly phosphorylated two residues (Ser515 and Ser518) within the Src homology 2 domain of JAK1. Activation of AMPK enhanced the interaction between JAK1 and 14-3-3 proteins in cultured vascular endothelial cells and fibroblasts, an effect that required the presence of Ser515 and Ser518 and was abolished in cells lacking AMPK catalytic subunits. Mutation of Ser515 and Ser518 abolished AMPK-mediated inhibition of JAK-STAT signaling stimulated by either the sIL-6Rα/IL-6 complex or the expression of a constitutively active V658F-mutant JAK1 in human fibrosarcoma cells. Clinically used AMPK activators metformin and salicylate enhanced the inhibitory phosphorylation of endogenous JAK1 and inhibited STAT3 phosphorylation in primary vascular endothelial cells. Therefore, our findings reveal a mechanism by which JAK1 function and inflammatory signaling may be suppressed in response to metabolic stress and provide a mechanistic rationale for the investigation of AMPK activators in a range of diseases associated with enhanced activation of the JAK-STAT pathway.

Here we demonstrate that overexpression of the human A(2A) adenosine receptor (A(2A)AR) in vascular endothelial cells confers an ability of interferon-alpha and a soluble IL-6 receptor/IL-6 (sIL-6R alpha/IL-6) trans-signaling complex to trigger the down-regulation of signal transducer and activator of transcription (STAT) proteins. It is noteworthy that STAT down-regulation could be reversed by coincubation with A(2A)AR-selective inverse agonist 4-(2-[7-amino-2-(2-furyl)[1,2,4]triazolo[2,3-a][1,3,5]triazin-5-ylamino]ethyl)phenol (ZM241385) but not adenosine deaminase, suggesting that constitutive activation of the receptor was responsible for the effect. Moreover, STAT down-regulation was selectively abolished by proteasome inhibitor N-benzoyloxycarbonyl (Z)-Leu-Leu-leucinal (MG132), whereas lysosome inhibitor chloroquine was without effect. Down-regulation required Janus kinase (JAK) activity and a Tyr705-->Phe-mutated STAT3 was resistant to the phenomenon, suggesting that JAK-mediated phosphorylation of this residue is required. Consistent with this hypothesis, treatment of A(2A)AR-overexpressing cells with sIL-6R alpha/IL-6 triggered the accumulation of polyubiquitylated wild-type but not Tyr705-->Phe-mutated STAT3. Support for a functional role of this process was provided by the observation that A(2A)AR overexpression attenuated the JAK/STAT-dependent up-regulation of vascular endothelial growth factor receptor-2 by sIL-6R alpha/IL-6. Consistent with a role for endogenous A(2A)ARs in regulating STAT protein levels, prolonged exposure of endogenous A(2A)ARs in endothelial cells with ZM241385 in vitro triggered the up-regulation of STAT3, whereas deletion of the A(2A)AR in vivo potentiated STAT1 expression and phosphorylation. Together, these experiments support a model whereby the A(2A)AR can prime JAK-phosphorylated STATs for polyubiquitylation and proteasomal degradation and represents a new mechanism by which an anti-inflammatory seven-transmembrane receptor can negatively regulate JAK/STAT signaling.

Inflammation of adipose tissue in obesity is associated with increased IL-1β, IL-6 and TNF-α secretion and proposed to contribute to insulin resistance. AMP-activated protein kinase (AMPK) regulates nutrient metabolism and is reported to have anti-inflammatory actions in adipose tissue, yet the mechanisms underlying this remain poorly characterised. The effect of AMPK activation on cytokine-stimulated proinflammatory signalling was therefore assessed in cultured adipocytes. AMPK activation inhibited IL-1β-stimulated CXCL10 secretion, associated with reduced interleukin-1 receptor associated kinase-4 (IRAK4) phosphorylation and downregulated MKK4/JNK and IKK/IκB/NFκB signalling. AMPK activation inhibited TNF-α-stimulated IKK/IκB/NFκB signalling but had no effect on JNK phosphorylation. The JAK/STAT3 pathway was also suppressed by AMPK after IL-6 stimulation and during adipogenesis. Adipose tissue from AMPKα1−/− mice exhibited increased JNK and STAT3 phosphorylation, supporting suppression of these distinct proinflammatory pathways by AMPK in vivo. The inhibition of multiple pro-inflammatory signalling pathways by AMPK may underlie the reported beneficial effects of AMPK activation in adipose tissue., Graphical abstract Image 1, Highlights • AMPK rapidly inhibits proinflammatory NFκB and JNK signalling in adipocytes. • Mechanisms underlying AMPK-induced effects involve inhibition of IRAK4, IKK and MKK4 phosphorylation. • AMPK concurrently inhibits JAK-STAT signalling in adipocytes. • Adipose tissue from mice lacking AMPKα1 exhibit increased proinflammatory signalling.

Here, we demonstrate that elevation of intracellular cyclic AMP (cAMP) in vascular endothelial cells (ECs) by either a direct activator of adenylyl cyclase or endogenous cAMP-mobilizing G protein-coupled receptors inhibited the tyrosine phosphorylation of STAT proteins by an interleukin 6 (IL-6) receptor trans-signaling complex (soluble IL-6Ralpha/IL-6). This was associated with the induction of suppressor of cytokine signaling 3 (SOCS-3), a bona fide inhibitor in vivo of gp130, the signal-transducing component of the IL-6 receptor complex. Attenuation of SOCS-3 induction in either ECs or SOCS-3-null murine embryonic fibroblasts abolished the inhibitory effect of cAMP, whereas inhibition of SHP-2, another negative regulator of gp130, was without effect. Interestingly, the inhibition of STAT phosphorylation and SOCS-3 induction did not require cAMP-dependent protein kinase activity but could be recapitulated upon selective activation of the alternative cAMP sensor Epac, a guanine nucleotide exchange factor for Rap1. Consistent with this hypothesis, small interfering RNA-mediated knockdown of Epac1 was sufficient to attenuate both cAMP-mediated SOCS-3 induction and inhibition of STAT phosphorylation, suggesting that Epac activation is both necessary and sufficient to observe these effects. Together, these data argue for the existence of a novel cAMP/Epac/Rap1/SOCS-3 pathway for limiting IL-6 receptor signaling in ECs and illuminate a new mechanism by which cAMP may mediate its potent anti-inflammatory effects.

Here we demonstrate that phosphorylation of the sphingosine-1-phosphate (S1P) receptor S1P 3 is increased specifically in response to S1P. Truncation of the receptor's carboxyl-terminal domain revealed that the presence of a serine-rich stretch of residues between Leu332 and Val352 was essential to observe this effect. Although agonist-occupied wild-type (WT) S1P 3 could be phosphorylated in vitro by G-protein-coupled receptor kinase 2 (GRK2), a role of S1P 3 phosphorylation in controlling S1P 3 –G q/11 coupling was excluded since A) a phosphorylation-resistant S1P 3 mutant desensitised in a manner indistinguishable from the WT receptor and was phosphorylated to a greater extent than the WT receptor by GRK2 in vitro, and B) co-expression with GRK2 or GRK3 failed to potentiate S1P 3 phosphorylation. S1P 3 phosphorylation was also not required for receptor sequestration away from the cell surface. Together, these data suggest that S1P 3 function is not subject to conventional regulation by GRK phosphorylation and that novel aspects of S1P 3 function distinct from classical G-protein coupling and receptor internalisation may be controlled its carboxyl-terminal domain.

Although the ability of bioactive lipid sphingosine-1-phosphate (S1P) to positively regulate anti-apoptotic/pro-survival responses by binding to S1P1 is well known, the molecular mechanisms remain unclear. Here we demonstrate that expression of S1P1 renders CCL39 lung fibroblasts resistant to apoptosis following growth factor withdrawal. Resistance to apoptosis was associated with attenuated accumulation of pro-apoptotic BH3-only protein Bim. However, although blockade of extracellular signal-regulated kinase (ERK) activation could reverse S1P1-mediated suppression of Bim accumulation, inhibition of caspase-3 cleavage was unaffected. Instead S1P1-mediated inhibition of caspase-3 cleavage was reversed by inhibition of phosphatidylinositol-3-kinase (PI3K) and protein kinase C (PKC), which had no effect on S1P1 regulation of Bim. However, S1P1 suppression of caspase-3 was associated with increased expression of anti-apoptotic protein Mcl-1, the expression of which was also reduced by inhibition of PI3K and PKC. A role for the induction of Mcl-1 in regulating endogenous S1P receptor-dependent pro-survival responses in human umbilical vein endothelial cells was confirmed using S1P receptor agonist FTY720-phosphate (FTY720P). FTY720P induced a transient accumulation of Mcl-1 that was associated with a delayed onset of caspase-3 cleavage following growth factor withdrawal, whereas Mcl-1 knockdown was sufficient to enhance caspase-3 cleavage even in the presence of FTY720P. Consistent with a pro-survival role of S1P1 in disease, analysis of tissue microarrays from ER+ breast cancer patients revealed a significant correlation between S1P1 expression and tumour cell survival. In these tumours, S1P1 expression and cancer cell survival were correlated with increased activation of ERK, but not the PI3K/PKB pathway. In summary, pro-survival/anti-apoptotic signalling from S1P1 is intimately linked to its ability to promote the accumulation of pro-survival protein Mcl-1 and downregulation of pro-apoptotic BH3-only protein Bim via distinct signalling pathways. However, the functional importance of each pathway is dependent on the specific cellular context.

The Janus kinase–signal transducer and activator of transcription (JAK–STAT) pathway is utilized by a range of cytokines (interferons, IL-2 and IL-6 amongst others) that control survival, proliferation and differentiation responses in diverse cell types. The realisation that unregulated activation of this pathway is a key driver of not only chronic inflammatory diseases such as rheumatoid arthritis, colitis and psoriasis, but also many cancers has identified its components as targets for therapeutic intervention by small molecule inhibitors and biologicals. In this article, we will discuss how an increased understanding of JAK-STAT pathway architecture, the basis for its dysfunction in pathological states, and its regulation by other intracellular signaling pathways are illuminating multiple strategies to manipulate this pathway in several disease arenas.

Prototypical second messenger cyclic AMP (cAMP) was originally thought to mediate its effects through activation of cAMP-dependent protein kinase (PKA). However, it is now clear that cells possess multiple alternative sensors of cAMP accumulation, of which the “exchange protein directly activated by cAMP” (Epac) proteins have been studied most intensively. This article will describe recent insights made into the molecular mechanisms by which Epac proteins mediate key protective effects of cAMP on two specific aspects of vascular endothelial cell function, namely barrier function and suppression of inflammatory signalling. It will also examine how integrative and unbiased global approaches are currently being deployed to answer several wider questions that have arisen from the identification of Epac as a trigger of gene transcription events and the E3 ubquitin ligase component “suppressor of cytokine signalling-3” (SOCS-3) as a key gene target regulated by this pathway.

Here we demonstrate that elevation of cyclic AMP (cAMP) levels in human umbilical vein endothelial cells (HUVECs) specifically attenuates ERK1,2 activation in response to either leptin or a soluble interleukin IL-6 receptor-alpha/IL-6 (sIL-6R alpha/IL-6) trans-signalling complex but not protein kinase C activator phorbol 12-myristate 13-acetate. The inhibitory effects of cAMP on sIL-6R alpha/IL-6-stimulated phosphorylation of ERK1,2 and STAT3 were abolished by either short interfering (si) RNA-mediated knockdown or genetic ablation of suppressor of cytokine signalling-3 (SOCS-3). The inhibitory effect of cAMP could not be reversed by inhibition of cAMP-dependent protein kinase (PKA) but was blocked by depletion of the alternative intracellular cAMP sensor exchange protein activated by cAMP 1 (Epac1), which is also required to observe SOCS-3 accumulation in response to cAMP. Interestingly, the ability of cAMP elevation to inhibit IL-6 signalling was blocked by ERK inhibition. Consistent with this observation, cAMP elevation in HUVECs produced a transient yet robust activation of ERK, and subsequent phosphorylation of transcription factor C/EBP beta, both of which were resistant to PKA inhibition. However, siRNA depletion and immunoblotting experiments revealed that neither Epac1 nor Epac2 contributed to the PKA-independent activation of ERK1,2 observed following cAMP elevation. Together, these observations suggest that while SOCS-3 induction and subsequent inhibition of cytokine-mediated phosphorylation of ERK1,2 and STAT3 in response to cAMP require Epac1 and a transient PKA-independent activation of the ERK pathway, these two events are controlled by distinct mechanisms. In addition, it reveals a novel Epac-dependent mechanism by which cAMP can specifically inhibit ERK in response to cytokine receptor activation.

patient records, nursing and medical notes. Data analysed using SPSS version 22. Results: Twenty-one patients with 26 TiLOOP assisted reconstructions with a mean follow up of 194 days (range 9e430) were analysed. Patients were a median age 51 (range 32e79) and median BMI of 25 (range 21e34). The majority had physical hobbies (n 1⁄4 16), no major comorbidities (n 1⁄4 17) and one was a smoker. The majority (n 1⁄4 19) stated the desire for a simple reconstructive option and were satisfied with their current breast size (n 1⁄4 18). 19 had in-situ or invasive breast cancer (n 1⁄4 13 screen-detected), 2 were risk reducing (proven BRCA mutation). Complications included: 1 haematoma; 5 seromas requiring single (40e150ml) aspiration; 3 with seroma treated as suspected minor infection with oral antibiotics. One implant loss was seen in this series. There was no correlation of complications with cup size, comorbidities or smoking status. Conclusions: TiLOOP is an economical non-autologous material available to provide lower-pole support and facilitate one-stage implantbased IBR, which is associated with a low complication rate.

patient records, nursing and medical notes. Data analysed using SPSS version 22. Results: Twenty-one patients with 26 TiLOOP assisted reconstructions with a mean follow up of 194 days (range 9e430) were analysed. Patients were a median age 51 (range 32e79) and median BMI of 25 (range 21e34). The majority had physical hobbies (n 1⁄4 16), no major comorbidities (n 1⁄4 17) and one was a smoker. The majority (n 1⁄4 19) stated the desire for a simple reconstructive option and were satisfied with their current breast size (n 1⁄4 18). 19 had in-situ or invasive breast cancer (n 1⁄4 13 screen-detected), 2 were risk reducing (proven BRCA mutation). Complications included: 1 haematoma; 5 seromas requiring single (40e150ml) aspiration; 3 with seroma treated as suspected minor infection with oral antibiotics. One implant loss was seen in this series. There was no correlation of complications with cup size, comorbidities or smoking status. Conclusions: TiLOOP is an economical non-autologous material available to provide lower-pole support and facilitate one-stage implantbased IBR, which is associated with a low complication rate.

Vitamin E is a potent, naturally occurring, lipid-soluble antioxidant, which is reported to be protective against several disease processes, including coronary atherosclerosis. We have measured the alpha-tocopherol content of the aorta, liver, skeletal muscle, and kidney of rats fed one of the following diets for 10 weeks: a normal control chow diet (i); or the same diet containing 1% cholesterol (ii); 0.5% vitamin E (iii); or 1% cholesterol plus 0.5% vitamin E (iv). The alpha-tocopherol content of serum and tissue extracts was measured by HPLC using gamma-tocopherol as an internal standard. Tissue and serum cholesterol content was measured using a cholesterol oxidase enzyme reagent kit. In all animals receiving the 1% cholesterol diet, serum cholesterol levels increased significantly (P < 0.005). By the 10th week, mean serum alpha-tocopherol levels rose significantly in both groups of animals receiving dietary vitamin E supplements (P < 0.0001) compared with their respective control group. This was accompanied by a significant increase in the absolute alpha-tocopherol content of liver (8- to 9-fold) and aorta (3- to 4-fold). The alpha-tocopherol content of renal and skeletal muscle tissue was raised 1- to 2-fold in both groups of rats on vitamin E supplements, however the increased attained significance only for the renal tissue. The aortic tissue alpha-tocopherol/cholesterol ratio was 4-fold higher in the rats receiving concomitant 1% cholesterol plus 0.5% vitamin E compared with animals receiving 1% cholesterol alone (P < 0.02), and was 5-fold higher in the rats receiving 0.5% vitamin E compared with those receiving control chow (P < 0.01). These data suggest that dietary vitamin E supplementation results in a differential uptake of alpha-tocopherol, which may be dependent, in part, on selective lipoprotein particle accumulation.