Proteinase-activated receptors, targets for kallikrein signaling

Serine proteinases like thrombin can signal to cells by the cleavage/activation of proteinase-activated receptors (PARs). Although thrombin is a recognized physiological activator of PAR(1) and PAR(4), the endogenous enzymes responsible for activating PAR(2) in settings other than the gastrointestinal system, where trypsin can activate PAR(2), are unknown. We tested the hypothesis that the human tissue kallikrein (hK) family of proteinases regulates PAR signaling by using the following: 1) a high pressure liquid chromatography (HPLC)-mass spectral analysis of the cleavage products yielded upon incubation of hK5, -6, and -14 with synthetic PAR N-terminal peptide sequences representing the cleavage/activation motifs of PAR(1), PAR(2), and PAR(4); 2) PAR-dependent calcium signaling responses in cells expressing PAR(1), PAR(2), and PAR(4) and in human platelets; 3) a vascular ring vasorelaxation assay; and 4) a PAR(4)-dependent rat and human platelet aggregation assay. We found that hK5, -6, and -14 all yielded PAR peptide cleavage sequences consistent with either receptor activation or inactivation/disarming. Furthermore, hK14 was able to activate PAR(1), PAR(2), and PAR(4) and to disarm/inhibit PAR(1). Although hK5 and -6 were also able to activate PAR(2), they failed to cause PAR(4)-dependent aggregation of rat and human platelets, although hK14 did. Furthermore, the relative potencies and maximum effects of hK14 and -6 to activate PAR(2)-mediated calcium signaling differed. Our data indicate that in physiological settings, hKs may represent important endogenous regulators of the PARs and that different hKs can have differential actions on PAR(1), PAR(2), and PAR(4).     

LKB1 in endothelial cells is required for angiogenesis and TGFbeta-mediated vascular smooth muscle cell recruitment

Inactivation of the tumor suppressor kinase Lkb1 in mice leads to vascular defects and midgestational lethality at embryonic day 9-11 (E9-E11). Here, we have used conditional targeting to investigate the defects underlying the Lkb1(-/-) phenotype. Endothelium-restricted deletion of Lkb1 led to embryonic death at E12.5 with a loss of vascular smooth muscle cells (vSMCs) and vascular disruption. Transforming growth factor beta (TGFbeta) pathway activity was reduced in Lkb1-deficient endothelial cells (ECs), and TGFbeta signaling from Lkb1(-/-) ECs to adjacent mesenchyme was defective, noted as reduced SMAD2 phosphorylation. The addition of TGFbeta to mutant yolk sac explants rescued the loss of vSMCs, as evidenced by smooth muscle alpha actin (SMA) expression. These results reveal an essential function for endothelial Lkb1 in TGFbeta-mediated vSMC recruitment during angiogenesis.     

Shrink and share: humanity's present and future Ecological Footprint

Sustainability is the possibility of all people living rewarding lives within the means of nature. Despite ample recognition of the importance of achieving sustainable development, exemplified by the Rio Declaration of 1992 and the United Nations Millennium Development Goals, the global economy fails to meet the most fundamental minimum condition for sustainability--that human demand for ecosystem goods and services remains within the biosphere's total capacity. In 2002, humanity operated in a state of overshoot, demanding over 20% more biological capacity than the Earth's ecosystems could regenerate in that year. Using the Ecological Footprint as an accounting tool, we propose and discuss three possible global scenarios for the future of human demand and ecosystem supply. Bringing humanity out of overshoot and onto a potentially sustainable path will require managing the consumption of food, fibre and energy, and maintaining or increasing the productivity of natural and agricultural ecosystems.     

Middle Triassic carbonate-platform break-up and formation of small-scale half-grabens (Julian and Kamnik–Savinja Alps, Slovenia)

In the Julian Alps (Mt. Prisojnik, NW Slovenia) and in the Kamnik–Savinja Alps (Mt. Kri?evnik, N Slovenia), both of which form part of the eastern Southern Alps, several meters of Upper Anisian pelagic red nodular, radiolarian-rich limestone (Loibl Formation) were deposited on the drowned platform carbonates of the Contrin Formation. The time of the platform drowning is dated with radiolarians and conodonts to the Illyrian, more precisely to the upper part of the Paraceratites trinodosus Ammonoid Zone. The red limestone is overlain by pyroclastics and volcanics (rhyolites) or carbonate (mega)breccia (Uggowitz Formation). The following unit consists of thin-bedded limestone, grainstone and subordinate marl (Buchenstein Formation) deposited during the final filling of the basin from the adjacent prograding carbonate platform (Schlern Formation) in the Ladinian. Map-scale geometry, neptunian dykes, the onset of volcanism, the presence of (mega)breccia and related paleo-escarpments, the lateral variations in thickness and the wedge-shaped geometry of the lithological units provide evidence of syn-sedimentary block faulting and the formation of small-scale, relatively shallow half-grabens within the previously uniform Slovenian Carbonate Platform. This analysis indicates a clear tectonic control over the development of the Middle Triassic stratigraphy. The described extensional event is well correlated and genetically connected with the syn-rift formation of the neighboring Slovenian Basin and other Southern Alpine basins that formed in connection with the opening of the Meliata-Maliac branch of the Neotethys Ocean.     

Human baby hair amino acid natural abundance 15N-isotope values are not related to the 15N-isotope values of amino acids in mother’s breast milk protein

Since exclusively breast-suckled infants obtain their nutrient only from their mother’s milk, it might be anticipated that a correlation will exist between the ¹⁵N/¹⁴N isotope ratios of amino acids of protein of young infants and those supplied by their mother. The work presented here aimed to determine whether amino nitrogen transfer from human milk to infant hair protein synthesized within the first month of life conserves the maternal isotopic signature or whether post-ingestion fractionation dominates the nitrogen isotope spectrum. The study was conducted at 1 month post-birth on 100 mother–infant pairs. Isotope ratios ¹⁵N/¹⁴N and ¹³C/¹²C were measured using isotope ratio measurement by Mass Spectrometry (irm-MS) for whole maternal milk, and infant hair and ¹⁵N/¹⁴N ratios were also measured by GC-irm-MS for the N-pivaloyl-O-isopropyl esters of amino acids obtained from the hydrolysis of milk and hair proteins. The δ¹⁵N and δ¹³C (‰) were found to be significantly higher in infant hair than in breast milk (δ¹⁵N, P < 0.001; δ¹³C, P < 0.001). Furthermore, the δ¹⁵N (‰) of individual amino acids in infant hair was also significantly higher than that in maternal milk (P < 0.001). By calculation, the observed shift in isotope ratio was shown not to be accounted for by the amino acid composition of hair and milk proteins, indicating that it is not simply due to differences in the composition in the proteins present. Rather, it would appear that each pool—mother and infant—turns over independently, and that fractionation in infant N-metabolism even in the first month of life dominates over the nutrient N-content.     

Treatment Success in Cancer: Industry Compared to Publicly Sponsored Randomized Controlled Trials

Objective

To assess if commercially sponsored trials are associated with higher success rates than publicly-sponsored trials.

Study Design and Settings

We undertook a systematic review of all consecutive, published and unpublished phase III cancer randomized controlled trials (RCTs) conducted by GlaxoSmithKline (GSK) and the NCIC Clinical Trials Group (CTG). We included all phase III cancer RCTs assessing treatment superiority from 1980 to 2010. Three metrics were assessed to determine treatment successes: (1) the proportion of statistically significant trials favouring the experimental treatment, (2) the proportion of the trials in which new treatments were considered superior according to the investigators, and (3) quantitative synthesis of data for primary outcomes as defined in each trial.

Results

GSK conducted 40 cancer RCTs accruing 19,889 patients and CTG conducted 77 trials enrolling 33,260 patients. 42% (99%CI 24 to 60) of the results were statistically significant favouring experimental treatments in GSK compared to 25% (99%CI 13 to 37) in the CTG cohort (RR = 1.68; p = 0.04). Investigators concluded that new treatments were superior to standard treatments in 80% of GSK compared to 44% of CTG trials (RR = 1.81; p<0.001). Meta-analysis of the primary outcome indicated larger effects in GSK trials (odds ratio = 0.61 [99%CI 0.47–0.78] compared to 0.86 [0.74–1.00]; p = 0.003). However, testing for the effect of treatment over time indicated that treatment success has become comparable in the last decade.

Conclusions

While overall industry sponsorship is associated with higher success rates than publicly-sponsored trials, the difference seems to have disappeared over time.     

Joint Loads in Marsupial Ankles Reflect Habitual Bipedalism versus Quadrupedalism

Joint surfaces of limb bones are loaded in compression by reaction forces generated from body weight and musculotendon complexes bridging them. In general, joints of eutherian mammals have regions of high radiodensity subchondral bone that are better at resisting compressive forces than low radiodensity subchondral bone. Identifying similar form-function relationships between subchondral radiodensity distribution and joint load distribution within the marsupial postcranium, in addition to providing a richer understanding of marsupial functional morphology, can serve as a phylogenetic control in evaluating analogous relationships within eutherian mammals. Where commonalities are established across phylogenetic borders, unifying principles in mammalian physiology, morphology, and behavior can be identified. Here, we assess subchondral radiodensity patterns in distal tibiae of several marsupial taxa characterized by different habitual activities (e.g., locomotion). Computed tomography scanning, maximum intensity projection maps, and pixel counting were used to quantify radiodensity in 41 distal tibiae of bipedal (5 species), arboreal quadrupedal (4 species), and terrestrial quadrupedal (5 species) marsupials. Bipeds (Macropus and Wallabia) exhibit more expansive areas of high radiodensity in the distal tibia than arboreal (Dendrolagus, Phascolarctos, and Trichosurus) or terrestrial quadrupeds (Sarcophilus, Thylacinus, Lasiorhinus, and Vombatus), which may reflect the former carrying body weight only through the hind limbs. Arboreal quadrupeds exhibit smallest areas of high radiodensity, though they differ non-significantly from terrestrial quadrupeds. This could indicate slightly more compliant gaits by arboreal quadrupeds compared to terrestrial quadrupeds. The observed radiodensity patterns in marsupial tibiae, though their statistical differences disappear when controlling for phylogeny, corroborate previously documented patterns in primates and xenarthrans, potentially reflecting inferred limb use during habitual activities such as locomotion. Despite the complex nature of factors contributing to joint loads, broad observance of these patterns across joints and across a variety of taxa suggests that subchondral radiodensity can be used as a unifying form-function principle within Mammalia.