Developmental profiles of GFAP-positive astrocytes in sheep cerebellum

Astroglial account for the largest glial population in the brain and play a variety of vital functions in the development of the central nervous system (CNS). An immunohistochemical study was performed in 19 ovine foetuses ranging from 2 to 5 months of gestation, one newborn lamb and three adult sheep. Using the anit-glial fibrillary acidic protein (GFAP) marker, several variations were found in the degree of GFAP positive (GFAP+) astrocyte distribution between the different zones in the cerebellum of sheep during brain development. Our study indicates that the first appearance of astrocytes from restricted zones in the cerebellum occurs around the eighth week of gestation. Bergmann cells were found to be present from around the 15th week of gestation onwards. Our findings suggest that the maturation of astrocytes begins in the caudal parts of the cerebellum, developing from their initial ventral regions to spread first to dorsal regions radially within the white matter, then followed by the more rostral parts of the cerebellum. Astrocytes were also found to proliferate in the vermis before appearing in the cerebellar hemispheres.     

A universal method to produce low-work function electrodes for organic electronics

Organic and printed electronics technologies require conductors with a work function that is sufficiently low to facilitate the transport of electrons in and out of various optoelectronic devices. We show that surface modifiers based on polymers containing simple aliphatic amine groups substantially reduce the work function of conductors including metals, transparent conductive metal oxides, conducting polymers, and graphene. The reduction arises from physisorption of the neutral polymer, which turns the modified conductors into efficient electron-selective electrodes in organic optoelectronic devices. These polymer surface modifiers are processed in air from solution, providing an appealing alternative to chemically reactive low-work function metals. Their use can pave the way to simplified manufacturing of low-cost and large-area organic electronic technologies.     

Using ecological niche modelling to infer past, present and future environmental suitability for Leiopelma hochstetteri, an endangered New Zealand native frog

Leiopelma hochstetteri is an endangered New Zealand frog now confined to isolated populations scattered across the North Island. A better understanding of its past, current and predicted future environmental suitability will contribute to its conservation which is in jeopardy due to human activities, feral predators, disease and climate change. Here we use ecological niche modelling with all known occurrence data (N = 1708) and six determinant environmental variables to elucidate current, pre-human and future environmental suitability of this species. Comparison among independent runs, subfossil records and a clamping method allow validation of models. Many areas identified as currently suitable do not host any known populations. This apparent discrepancy could be explained by several non exclusive hypotheses: the areas have not been adequately surveyed and undiscovered populations still remain, the model is over simplistic; the species’ sensitivity to fragmentation and small population size; biotic interactions; historical events. An additional outcome is that apparently suitable, but frog-less areas could be targeted for future translocations. Surprisingly, pre-human conditions do not differ markedly highlighting the possibility that the range of the species was broadly fragmented before human arrival. Nevertheless, some populations, particularly on the west of the North Island may have disappeared as a result of human mediated habitat modification. Future conditions are marked with higher temperatures, which are predicted to be favourable to the species. However, such virtual gain in suitable range will probably not benefit the species given the highly fragmented nature of existing habitat and the low dispersal ability of this species.     

Vasomodulating potential of mediterranean wild plant extracts

The incidence of cardiovascular disease and endothelial dysfunction is low in the Mediterranean area, where the major proportion of daily calories comes from plant food, high in antioxidant polyphenols. It has been shown that a reduced production or enhanced inactivation of endothelium-derived nitric oxide (NO) is involved in the onset of endothelial dysfunction. We investigated the effects of Mediterranean wild plant, that is, wild artichoke and thyme, phenolic-rich extracts on NO release by porcine aortic endothelial cells (PAECs; by using indirect methods) and by cerebral cell membrane homogenates (by using direct NO detection). NO release by PAECs was significantly potentiated by 234% and 135% by wild artichoke and thyme extracts (10(-6) mol/L), respectively. Direct detection of NO release by brain membranes also showed significantly increased NO production after wild artichoke addition (+35.4%). Further, the release of another vasorelaxant factor by PAECs, that is, prostacyclin, was significantly increased by wild artichoke and thyme (10(-6) mol/L) (+269% and +190%, respectively). Investigation of the mechanism(s) of action of wild artichoke and thyme suggests maintenance of an intracellular reduced environment, as previously shown for ascorbate. Even though these data require in vivo confirmation, they suggest that regular intake of bioactive compounds from Mediterranean wild plants contributes to maintenance of proper vasomotion and to the low incidence of atherosclerosis and endothelial dysfunction recorded in the Mediterranean area.     

Pharmacokinetics of low-dose methotrexate in horses

This study aimed to investigate both the pharmacokinetic behavior and tolerance of methotrexate (MTX) in horses to design a specific dosing regimen as a new immunomodulatory drug for long-term treatment. To determine the primary plasma pharmacokinetic variables after single intravenous, subcutaneous or oral administration, six horses were administered 0.3 mg/kg MTX in a crossover design study. After a 10-week washout, MTX was administered subcutaneously to three of the six previously treated horses at a dose of 0.3 mg/kg once per week for 3 months. In both studies, MTX and metabolite concentrations were measured using LC-MS/MS. The absolute bioavailability of MTX was 73% following subcutaneous administration but less than 1% following oral administration. The plasma clearance was 1.54 ml min⁻¹ kg⁻¹ (extraction ratio = 2%). After 24 hr, plasma concentrations were below the LOQ. No adverse effects were noted except for a moderate reversible elevation in liver enzymes (GLDH). With regards to the main metabolites of MTX, very low concentrations of 7-hydroxy-MTX were found, whereas polyglutamated forms (mainly short chains) were found in red blood cells. A subcutaneous dose of 0.2 mg kg⁻¹ week⁻¹ may be safe and relevant in horses, although this has yet to be clinically confirmed.     

Rein tension acceptance in young horses in a voluntary test situation

Reasons for performing the study: During riding, horses are frequently exposed to pressure from the rider, e.g. through the bit and reins, but few studies have investigated at which point rein tension becomes uncomfortable for the horse. Objectives: To investigate how much rein tension young inexperienced horses are willing to accept in order to obtain a food reward; whether the tension acceptance changes during 3 consecutive test days; and whether rein tension correlates with the expression of conflict behaviour and heart rate. Hypotheses: Pressure‐naïve horses will apply only little rein tension in the first voluntary trial, but their acceptance will gradually increase. High levels of rein tension will lead to expression of conflict behaviour and increases in heart rate. Methods: Fifteen 2‐year‐old, bridle‐naïve mares were encouraged to stretch their head forward (across a 0.95 m high metal bar) to obtain a food reward in a voluntary test situation. On each test day, each horse was exposed to 2 control sessions (loose reins), an intermediate and a short rein session (1 min/session). Rein tension, heart rate and behaviour were recorded. Results: The horses applied significantly more tension on the first day (mean rein tension: 10.2 N), compared to the second and third test day (Day 2: 6.0 and Day 3: 5.7 N). The horses showed significantly more conflict behaviour in the short rein treatment. There was no treatment effect on heart rate. Conclusions and potential relevance: The horses applied the highest rein tension on the first day, and apparently learned to avoid the tension, rather than habituate to it. Rein tension correlated with expression of conflict behaviour, indicating that the horses found the tension aversive. Further studies should focus on the correlation between rein tension and conflict behaviour in ridden horses.     

A conserved mutation in an ethylene biosynthesis enzyme leads to andromonoecy in melons

Andromonoecy is a widespread sexual system in angiosperms characterized by plants carrying both male and bisexual flowers. In melon, this sexual form is controlled by the identity of the alleles at the andromonoecious (a) locus. Cloning of the a gene reveals that andromonoecy results from a mutation in the active site of 1-aminocyclopropane-1-carboxylic acid synthase. Expression of the active enzyme inhibits the development of the male organs and is not required for carpel development. A causal single-nucleotide polymorphism associated with andromonoecy was identified, which suggests that the a allele has been under recent positive selection and may be linked to the evolution of this sexual system.     

A simple test to estimate the salinity resistance of fish with specific application to O. niloticus and S. melanotheron

In order to develop a simple and accurate index of the salinity resistance of tilapia, batches of 10 juveniles (5 to 20 g) of two different species Oreochromis niloticus and Sarotherodon melanotheron reared in freshwater were subjected to gradual increases in salinity until 100% mortality. Seven daily increments of salinity were tested with 4 replicates: 2, 4, 6, 8, 10, 12 and 14 g l⁻¹ day⁻¹, while control batches were kept in fresh water. The temperature was maintained at 27 °C. The concentration of oxygen, ammonia and the pH were not limiting factors. The mortality, monitored on a daily basis, appeared after 2–51 days and was spread out over 1–20 days, depending on the increment of salinity. The higher the daily rate in salinity increase, then the shorter the time lapse before total mortality occurred. The cumulative mortality as a function of salinity fit well with simple linear regressions. The criterion of the resistance to salinity was the index MLS (median lethal salinity) defined at each daily rate as the salinity at which 50% of fish died. For S. melanotheron, the mean MLS was 123.7±3.5 g l⁻¹ whatever the daily rate in salinity. For O. niloticus, the MLS was 46.3±3.4 g l⁻¹ for daily increases in salinity ranging from 2 to 8 g l⁻¹ day⁻¹ and decreased significantly (P<0.05) above this level. The MLS-8 g l⁻¹ day⁻¹ ,which takes into account the full capacity of the fish to adapt to the increasing salinity, appeared to be a simple, optimized and efficient criterion for assessing the resistance to salinity for O. niloticus and S. melanotheron. This criterion can be a useful tool for ranking the different parental strains and hybrids of different genus and species of tilapia used in programmes of genetic selection for growth and salinity tolerance.     

Diffusion dynamics of glycine receptors revealed by single-quantum dot tracking

Semiconductor quantum dots (QDs) are nanometer-sized fluorescent probes suitable for advanced biological imaging. We used QDs to track individual glycine receptors (GlyRs) and analyze their lateral dynamics in the neuronal membrane of living cells for periods ranging from milliseconds to minutes. We characterized multiple diffusion domains in relation to the synaptic, perisynaptic, or extrasynaptic GlyR localization. The entry of GlyRs into the synapse by diffusion was observed and further confirmed by electron microscopy imaging of QD-tagged receptors.     

Wheat bran tissue fractionation using biochemical markers

Phenolic acid analysis of hand-isolated outer grain layers and endosperm led to the identification of markers of pericarp and aleurone layers, respectively. A new dehydrotrimer of ferulic acid (DHT) was found to be concentrated in the outer pericarp of wheat bran whereas p-coumaric (p-CA) acid was mainly in the aleurone layer. Phytates were also used as a marker of aleurone layer and starch as a marker of starchy endosperm. Biochemical markers constitute an original method for determining the histological composition of any technological bran fractions. A pin milling process was applied to coarse bran produced by a conventional milling process. Three different fractions (B1, B2 and B3) were obtained by sieving the bran products and then the smallest bran particle fraction (B3) was air-classified to obtain two particle size fractions (B3a and B3b with a D50 of 83 and 7 μm, respectively). The biochemical composition of these fractions was used to calculate the distribution of tissues according to the sieving process. The dissociation behavior of individual bran tissues upon mechanical fractionation was investigated in relation to particle size and discussed according to their mechanical properties.