Optimal oxidative folding of the novel antimicrobial cyclotide from Hedyotis biflora requires high alcohol concentrations

Hedyotide B1, a novel cyclotide isolated from the medicinal plant Hedyotis biflora, contains a cystine knot commonly found in toxins and plant defense peptides. The optimal oxidative folding of a cystine knot encased in the circular peptide backbone of a cyclotide poses a challenge. Here we report a systematic study of optimization of the oxidative folding of hedyotide B1, a 30-amino acid cyclic peptide with a net charge of +3. The linear precursor of hedyotide B1, synthesized as a thioester by solid phase synthesis, was cyclized quantitatively by a thia-zip cyclization to form the circular backbone and then subjected to oxidative folding in a thiol-disulfide redox system under 38 different conditions. Of the oxidative conditions examined, the nature of the organic cosolvent appeared to be critical, with the use of 70% 2-propanol affording the highest yield (48%). The disulfide connectivity of the folded hedyotide was identical to that of the native form as determined by partial acid hydrolysis. The use of such a high alcohol concentration suggests that a partial denaturation may be necessary for the oxidative folding of a cyclotide with the inverse orientation of hydrophobic side chains that are externalized to the solvent face to permit the formation of the interior cystine core in the circularized backbone. We also show that synthetic hedyotide B1 is an antimicrobial, exhibiting minimal inhibitory concentrations in the micromolar range against both Gram-positive and -negative bacteria.     

Impact of WNT signaling on tissue lineage differentiation in the early mouse embryo

WNT signaling activity is involved in the regulation of many cellular functions, including proliferation, migration, cell fate specification, maintenance of pluripotency and induction of tumorigenicity. Here we summarize recent progress towards understanding the regulation of canonical WNT/β-catenin signaling activity through feedback regulatory loops involving the ligands, agonists and antagonists, the availability of intracellular pools of active β-catenin and the cross-regulation of the WNT activity by β-catenin independent pathway. We also review recent findings on the role of WNT/β-catenin signaling in tissue lineage differentiation during embryogenesis and the maintenance and self renewal of embryo-derived stem cells in vitro.     

Genetic structure among Charadrius plovers on the African mainland and islands of Madagascar and St Helena

Colonization of islands by long-distance dispersers has great impact on genetic diversification among populations and may spearhead speciation events. We investigated intra- and interspecific divergence in Charadrius plovers with populations on mainland Africa, Madagascar and St Helena. We analysed microsatellite loci and sequence data from four nuclear and two mitochondrial gene regions. Charadrius plovers are shorebirds with high dispersal and mobility. Our results confirmed genetic differentiation between Madagascar and mainland populations of three plover species (White-fronted Plover Charadrius marginatus, Kittlitz's Plover Charadrius pecuarius and, based on sequence data only, Three-banded Plover Charadrius tricollaris) but highlight substantial variation in levels of intraspecific divergence among the three species. Namely, the Kittlitz's Plover, a dispersive habitat generalist with a polygamous mating system, exhibited lower island–mainland differentiation (0.05% COI sequence divergence) compared with the two monogamous species, the White-fronted Plover (0.6% COI divergence) and Three-banded Plover (1.6% COI divergence). In addition, past colonization of the islands of St Helena and Madagascar by ancestors of today's Kittlitz's Plover has led to the evolution of two endemic island species, the Madagascar Plover Charadrius thoracicus and the more closely-related St Helena Plover Charadrius sanctaehelenae. We discuss the factors driving species differences in island–mainland divergence and highlight the importance of conserving genetically unique island populations and island habitats to safeguard future evolutionary potential.     

Estimates of Dengue Force of Infection in Children in Colombo,Sri Lanka

Dengue is the most important vector-borne viral disease worldwide and a major cause of childhood fever burden in Sri Lanka, which has experienced a number of large epidemics in the past decade. Despite this, data on the burden and transmission of dengue virus in the Indian Subcontinent are lacking. As part of a longitudinal fever surveillance study, we conducted a dengue seroprevalence survey among children aged <12 years in Colombo, Sri Lanka. We used a catalytic model to estimate the risk of primary infection among seronegative children. Over 50% of children had IgG antibodies to dengue virus and seroprevalence increased with age. The risk of primary infection was 14.1% per year (95% CI: 12.7%–15.6%), indicating that among initially seronegative children, approximately 1 in 7 experience their first infection within 12 months. There was weak evidence to suggest that the force of primary infection could be lower for children aged 6 years and above. We estimate that there are approximately 30 primary dengue infections among children <12 years in the community for every case notified to national surveillance, although this ratio is closer to 100∶1 among infants. Dengue represents a considerable infection burden among children in urban Sri Lanka, with levels of transmission comparable to those in the more established epidemics of Southeast Asia.     

Unintended Consequences of Invasive Predator Control in an Australian Forest: Overabundant Wallabies and Vegetation Change

Over-abundance of native herbivores is a problem in many forests worldwide. The abundance of native macropod wallabies is extremely high at Booderee National Park (BNP) in south-eastern Australia. This has occurred because of the reduction of exotic predators through an intensive baiting program, coupled with the absence of other predators. The high density of wallabies at BNP may be inhibiting the recruitment of many plant species following fire-induced recruitment events. We experimentally examined the post-fire response of a range of plant species to browsing by wallabies in a forest heavily infested with the invasive species, bitou bush Chrysanthemoides monilifera. We recorded the abundance and size of a range of plant species in 18 unfenced (browsed) and 16 fenced (unbrowsed) plots. We found the abundance and size of bitou bush was suppressed in browsed plots compared to unbrowsed plots. Regenerating seedlings of the canopy or middle storey tree species Eucalyptus pilularis, Acacia implexa, Allocasuarina littoralis, Breynia oblongifolia and Banksia integrifolia were either smaller or fewer in number in grazed plots than treatment plots as were the vines Kennedia rubicunda, Glycine tabacina and Glycine clandestina. In contrast, the understorey fern, Pteridium esculentum increased in abundance in the browsed plots relative to unbrowsed plots probably because of reduced competition with more palatable angiosperms. Twelve months after plots were installed the community structure of the browsed and unbrowsed plots was significantly different (P = 0.023, Global R = 0.091). The relative abundance of C. monilifera and P. esculentum contributed most to the differences. We discuss the possible development of a low diversity bracken fern parkland in Booderee National Park through a trophic cascade, similar to that caused by overabundant deer in the northern hemisphere. We also discuss its implications for broad scale fox control in southern Australian forests.     

Nicotinamide N-methyltransferase expression in SH-SY5Y neuroblastoma and N27 mesencephalic neurones induces changes in cell morphology via ephrin-B2 and Akt signalling

Nicotinamide N-methyltransferase (NNMT, E.C. 2.1.1.1) N-methylates nicotinamide to produce 1-methylnicotinamide (MeN). We have previously shown that NNMT expression protected against neurotoxin-mediated cell death by increasing Complex I (CxI) activity, resulting in increased ATP synthesis. This was mediated via protection of the NDUFS3 subunit of CxI from degradation by increased MeN production. In the present study, we have investigated the effects of NNMT expression on neurone morphology and differentiation. Expression of NNMT in SH-SY5Y human neuroblastoma and N27 rat mesencephalic dopaminergic neurones increased neurite branching, synaptophysin expression and dopamine accumulation and release. siRNA gene silencing of ephrin B2 (EFNB2), and inhibition of Akt phosphorylation using {"type":"entrez-nucleotide","attrs":{"text":"LY294002","term_id":"1257998346","term_text":"LY294002"}}LY294002, demonstrated that their sequential activation was responsible for the increases observed. Incubation of SH-SY5Y with increasing concentrations of MeN also increased neurite branching, suggesting that the effects of NNMT may be mediated by MeN. NNMT had no significant effect on the expression of phenotypic and post-mitotic markers, suggesting that NNMT is not involved in determining phenotypic fate or differentiation status. These results demonstrate that NNMT expression regulates neurone morphology in vitro via the sequential activation of the EFNB2 and Akt cellular signalling pathways.     

Structural and Functional Divergence of Growth Hormone-Releasing Hormone Receptors in Early Sarcopterygians: Lungfish and Xenopus

The evolutionary trajectories of growth hormone-releasing hormone (GHRH) receptor remain enigmatic since the discovery of physiologically functional GHRH-GHRH receptor (GHRHR) in non-mammalian vertebrates in 2007. Interestingly, subsequent studies have described the identification of a GHRHR₂ in chicken in addition to the GHRHR and the closely related paralogous receptor, PACAP-related peptide (PRP) receptor (PRPR). In this article, we provide information, for the first time, on the GHRHR in sarcopterygian fish and amphibians by the cloning and characterization of GHRHRs from lungfish (P. dolloi ) and X. laevis. Sequence alignment and phylogenetic analyses demonstrated structural resemblance of lungfish GHRHR to their mammalian orthologs, while the X. laevis GHRHR showed the highest homology to GHRHR₂ in zebrafish and chicken. Functionally, lungfish GHRHR displayed high affinity towards GHRH in triggering intracellular cAMP and calcium accumulation, while X. laevis GHRHR₂ was able to react with both endogenous GHRH and PRP. Tissue distribution analyses showed that both lungfish GHRHR and X. laevis GHRHR₂ had the highest expression in brain, and interestingly, X. laevis _GHRHR2 also had high abundance in the reproductive organs. These findings, together with previous reports, suggest that early in the Sarcopterygii lineage, GHRHR and PRPR have already established diverged and specific affinities towards their cognate ligands. GHRHR₂, which has only been found in xenopus, zebrafish and chicken hitherto, accommodates both GHRH and PRP.     

Hierarchical Self-Organization of Non-Cooperating Individuals

Hierarchy is one of the most conspicuous features of numerous natural, technological and social systems. The underlying structures are typically complex and their most relevant organizational principle is the ordering of the ties among the units they are made of according to a network displaying hierarchical features. In spite of the abundant presence of hierarchy no quantitative theoretical interpretation of the origins of a multi-level, knowledge-based social network exists. Here we introduce an approach which is capable of reproducing the emergence of a multi-levelled network structure based on the plausible assumption that the individuals (representing the nodes of the network) can make the right estimate about the state of their changing environment to a varying degree. Our model accounts for a fundamental feature of knowledge-based organizations: the less capable individuals tend to follow those who are better at solving the problems they all face. We find that relatively simple rules lead to hierarchical self-organization and the specific structures we obtain possess the two, perhaps most important features of complex systems: a simultaneous presence of adaptability and stability. In addition, the performance (success score) of the emerging networks is significantly higher than the average expected score of the individuals without letting them copy the decisions of the others. The results of our calculations are in agreement with a related experiment and can be useful from the point of designing the optimal conditions for constructing a given complex social structure as well as understanding the hierarchical organization of such biological structures of major importance as the regulatory pathways or the dynamics of neural networks.