Generation of a nitric oxide signaling pathway in mesenchymal stem cells promotes endothelial lineage commitment

Enhancing differentiation of mesenchymal stem cells (MSCs) to endothelial cells may improve their ability to vascularize tissue and promote wound healing. This study describes a novel role for nitric oxide (NO) in reprogramming MSCs towards an endothelial lineage and highlights the role of Wnt signaling and epigenetic modification by NO. Rat MSCs were transduced with lentiviral vectors expressing endothelial nitric oxide synthase (pLV-eNOS) and a mutated caveolin gene (pLV-CAV-1^F92A) to enhance NO generation resulting in increased in vitro capillary tubule formation and endothelial marker gene expression. An exogenous source of NO could also stimulate CD31 expression in MSCs. NO was associated with an arterial-specific endothelial gene expression profile of Notch1, Dll4, and Hey2 and significantly reduced expression of venous markers. Wnt signaling associated with NO was evident through increased gene expression of Wnt3a and β-catenin protein, and expression of the endothelial marker Pecam-1 could be significantly reduced by treatment with the Wnt signaling inhibitor Dkk-1. The role of NO as an epigenetic modifier was evident with reduced gene expression of the methyltransferase, DNMT1, and bisulfite sequencing of the endothelial Flt1 promoter region in NO-producing MSCs showed significant demethylation compared to control cells. Finally, subcutaneous implantation of NO-producing MSCs seeded in a biomaterial scaffold (NovoSorb®) resulted in survival of transplanted cells and the formation of blood vessels. In summary, this study describes, NO as a potent endothelial programming factor which acts as an epigenetic modifier in MSCs and may provide a novel platform for vascular regenerative therapy.     

Contribution of K99 and D319 to substrate binding and catalysis in the saccharopine dehydrogenase reaction

Saccharopine dehydrogenase catalyzes the NAD-dependent oxidative deamination of saccharopine to l-lysine and α-ketoglutarate. Lysine 99 is within hydrogen-bond distance to the α-carboxylate of the lysine substrate and D319 is in the vicinity of the carboxamide side chain of NADH. Both are conserved and may be important to the overall reaction. Replacing K99 with M gives decreases of 110-, 80- and 20-fold in the V(2)/K(m) values for lysine, α-ketoglutarate and NADH, respectively. Deuterium isotope effects on V and V/K(Lys) increase, while the solvent deuterium isotope effects decrease compared to the C205S mutant enzyme. Data for K99M suggest a decreased affinity for all reactants and a change in the partition ratio of the imine intermediate to favor hydrolysis. A change in the bound conformation of the imine and/or the distance of the imine carbon to C4 of the nicotinamide ring of NADH is also suggested. Changing D319 to A decreases V(2)/K(NADH) by 33-fold. Primary deuterium and solvent deuterium isotope effects decrease compared to C205S suggesting a non-isotope sensitive step has become slower. NADH binds to enzyme first, and sets the site for binding of lysine and α-ketoglutarate. The slower step is likely the conformational change generated upon binding of NADH.     

Soil and vegetation responses to hydrological manipulation in a partially drained polje fen in New Zealand

Anthropogenic drainage causes loss of natural character in herbaceous wetlands due to increased soil oxygen penetration. We related vegetation gradients in a New Zealand polje fen to long-term effects of drains by using hydrological, edaphic and vegetation data, and a before-after-control-impact (BACI) design to test responses to experimental drain closure. Soil profiles and continuous water level records revealed a site subject to frequent disturbance by intense but brief floods, followed by long drying periods during which areas close to drains experienced lower water tables and more variable water levels. Classification of vegetation data identified 12 groups along a moisture gradient, from dry areas dominated by pastoral alien species, to wet communities dominated by native wetland sedges. Lower total species diversity and native representation in pastoral communities were related to the high proportion of alien competitor and competitor-disturbance species, compared with the stress tolerator-dominated flora of other groups. Species–environment relationships revealed highly significant correlations with soil water content and aeration as measured by redox potential (E_H) and steel rod oxidation depth, as well as soil nutrient content and bulk density. Comparison of root anatomy confirmed greater development of flood-tolerant traits in native species than in pastoral aliens, and vegetation N:P ratios indicated that most communities were probably nitrogen-limited. Flooding rapidly re-established wetland hydrology in dried sites in the impact area, and lowered E_H and soil oxidation depth, but had no effect on N and P availability. Presence and cover of pastoral alien species decreased in these areas. This study supports the use of hydrological manipulation as a tool for reducing soil oxidation and thus the impact of alien plant species at restoration sites with minimal intervention, but suggests the need for detailed information on species flooding tolerances and hydrological preferences to underpin this approach.     

Feeding patterns revealed host partitioning in a community of frog-biting mosquitoes

1. Resource partitioning is a critical component in competing species that coexist in a community. The biting behaviour of coexisting frog-biting mosquito species associated with a tropical anuran community is investigated. 2. Monthly samplings were taken for 2 years at two study sites in central Sri Lanka to collect frog-biting mosquitoes, anuran abundance, environmental data, and interactions between mosquitoes and anuran hosts. Mosquitoes were collected using mouth-operated aspirators and sound traps broadcasting anuran calls. Mosquitoes were identified using taxonomic keys and DNA barcodes. 3. A total of 1079 frog-biting mosquitoes from four species belonging to two genera were collected [Uranotaenia rutherfordi (5%), Ur. morphotype 1 (67%), Ur. morphotype 2 (21%), and Mansonia uniformis (7%)]. Species-specific interactions between Uranotaenia mosquitoes and their anuran host were found. Uranotaenia morphotype 1, the most common species, was mainly attracted (99%) to Duttaphrynus melanostictus. Uranotaenia rutherfordi was mainly attracted (95%) to Pseudophilautus rus, while Ur. morphotype 2 was attracted (97%) to Polypedates cruciger. These Uranotaenia species are active at different hours at night that correspond to the peak calling activity of their anuran host. Each Uranotaenia species was active at heights that coincide with the calling sites of their host. In contrast, Ma. uniformis was non-specific in host choice and was equally distributed in space and time with respect to host feeding. 4. Here, previously unknown feeding patterns of co-occurring frog-biting mosquito species and their interactions with anurans present in their community are reported, highlighting the existence of complex behavioural patterns of these mosquito communities.     

Metabolic profiling of benzoxazinoids in the roots and rhizosphere of commercial winter wheat genotypes

Plant and Soil - Integrated weed management in commercial wheat production is urgently needed due to increasing herbicide resistance and production costs. Benzoxazinoids (BXs), which include...     

Corvids congregate to breeding colonies of a burrow‐nesting seabird

Egg predation is a major cause of reproductive failure among birds, and can compromise the viability of affected populations. Some egg predators aggregate near colonially breeding birds to exploit the seasonal increase of prey resources. We investigated spatial and temporal variations in the abundance of an egg predator (little raven Corvus mellori; Corvidae) to identify whether ravens aggregate spatially or temporally to coincide with any of three potential prey species: burrow‐nesting little penguin (Eudyptula minor; Spheniscidae), short‐tailed shearwater (Ardenna tenuirostris; Procellariidae), and surface‐nesting silver gull (Chroicocephalus novaehollandiae; Laridae). We derived spatially explicit density estimates of little ravens using distance sampling along line transects throughout a calendar year, which encompassed little penguin, short‐tailed shearwater and silver gull breeding and non‐breeding seasons. High raven abundance coincided temporally with penguin and gull egg laying periods but not with that of shearwaters. The spatial distribution of raven density corresponded with the little penguin colony but not with shearwater or gull colonies. Thus, the presence of little penguin eggs in burrows correlated strongly with little raven activity, and this implies that little ravens may have learnt to exploit the plentiful subsurface food resource of little penguin eggs. Corvid management may be required to maintain the viability of this socially and economically important penguin colony.     

Frequency of impulse control behaviours associated with dopaminergic therapy in restless legs syndrome

Background  

Low doses of dopamine agonists (DA) and levodopa are effective in the treatment of restless legs syndrome (RLS). A range of impulse control and compulsive behaviours (ICBs) have been reported following the use of DAs and levodopa in patients with Parkinson's disease. With this study we sought to assess the cross-sectional prevalence of impulse control behaviours (ICBs) in restless legs syndrome (RLS) and to determine factors associated with ICBs in a population cohort in Germany.     

Visual behaviour of neonata larvae of the light brown apple moth

When neonate larvae of a leafroller moth,Epiphyas postvittana (Walker) (Lepidoptera: Tortricidae) were released into the middle of a circular arena with blue paper on one side of the arena and apple foliage on the other side, more larvae walked towards the apple foliage. These oriented responses were enhanced, in terms of the number of larvae responding, by increasing the amount of light reflected from or transmitted through apple foliage. Larvae also responded to painted targets, and specifically to targets reflecting light in the region of 470–570 nm (green-yellow region to the human eye). When the amount of 470–570 nm reflected from targets was reduced, numbers of larvae responding to targets decreased. The addition of 400–500 nm reflected light to 470–570 nm reflected light also resulted in a reduction of oriented responses to targets. Tests using neutral colours (white, black, and a series of greys) indicated that, in the absence of targets reflecting primarily in the 470–570 nm region, larvae oriented towards targets with low levels of reflectance. When the orientation of walking larvae was measured at various distances from targets of different colours or diameters, targets subtending 5–8 degrees elicited responses from 50% of all larvae. Behaviour other than walking was also influenced by visual stimuli: fewer larvae spun down on a silken thread when blue paper was placed beneath a walking platform than when brown or green papers were present.