RAF kinase activity regulates neuroepithelial cell proliferation and neuronal progenitor cell differentiation during early inner ear development
Maria Rodriguez Aburto
Ulf Rüdiger Rapp
- Background: Early inner ear development requires the strict regulation of cell proliferation, survival, migration and differentiation, coordinated by the concerted action of extrinsic and intrinsic factors. Deregulation of these processes is associated with embryonic malformations and deafness. We have shown that insulin-like growth factor I (IGF-I) plays a key role in embryonic and postnatal otic development by triggering the activation of intracellular lipid and protein kinases. RAF kinases are serine/threonine kinases that regulate the highly conserved RAS-RAF-MEK-ERK signaling cascade involved in transducing the signals from extracellular growth factors to the nucleus. However, the regulation of RAF kinase activity by growth factors during development is complex and still not fully understood.
Methodology/Principal Findings: By using a combination of qRT-PCR, Western blotting, immunohistochemistry and in situ hybridization, we show that C-RAF and B-RAF are expressed during the early development of the chicken inner ear in specific spatiotemporal patterns. Moreover, later in development B-RAF expression is associated to hair cells in the sensory patches. Experiments in ex vivo cultures of otic vesicle explants demonstrate that the influence of IGF-I on proliferation but not survival depends on RAF kinase activating the MEK-ERK phosphorylation cascade. With the specific RAF inhibitor Sorafenib, we show that blocking RAF activity in organotypic cultures increases apoptosis and diminishes the rate of cell proliferation in the otic epithelia, as well as severely impairing neurogenesis of the acoustic-vestibular ganglion (AVG) and neuron maturation.
Conclusions/Significance: We conclude that RAF kinase activity is essential to establish the balance between cell proliferation and death in neuroepithelial otic precursors, and for otic neuron differentiation and axonal growth at the AVG.
Transfer entropy - a model-free measure of effective connectivity for the neurosciences
- Understanding causal relationships, or effective connectivity, between parts of the brain is of utmost importance because a large part of the brain’s activity is thought to be internally generated and, hence, quantifying stimulus response relationships alone does not fully describe brain dynamics. Past efforts to determine effective connectivity mostly relied on model based approaches such as Granger causality or dynamic causal modeling. Transfer entropy (TE) is an alternative measure of effective connectivity based on information theory. TE does not require a model of the interaction and is inherently non-linear. We investigated the applicability of TE as a metric in a test for effective connectivity to electrophysiological data based on simulations and magnetoencephalography (MEG) recordings in a simple motor task. In particular, we demonstrate that TE improved the detectability of effective connectivity for non-linear interactions, and for sensor level MEG signals where linear methods are hampered by signal-cross-talk due to volume conduction.
Prevalence of dosing errors in elderly patients with impaired renal function: a survey in ambulatory patients
- Meeting Abstract : Gesellschaft für Arzneimittelanwendungsforschung und Arzneimittelepidemiologie e.V. (GAA). 17. Jahrestagung der Gesellschaft für Arzneimittelanwendungsforschung und Arzneimittelepidemiologie. Osnabrück, 25.-26.11.2010.
ntroduction: Several drugs require dose adjustment in patients with impaired renal function, which however, often goes undetected. Serum creatinine may be normal in patients while renal function is already reduced. The estimated GFR (eGFR) allows a more precise evaluation of the renal function. This study was carried out in a group practice for family medicine, in Frankfurt/ Main, Germany. The exploration aimed at investigating if patients with renal insufficiency were recognised and if their prescriptions were appropriate in terms of dose adjustment or contra-indications.
Methods: In patients (>65yrs) with renal insufficiency (creatinine clearance <60 ml/min), their prescribed medication was retrospectively explored (Observation period 1.1.2008 to 1.4.2009). The Cockroft-Gault formula was used as estimate for the eGFR, using a creatinine value from the patient’s charts. In 90 patients, a second eGFR could be estimated from a second creatinine value obtained within 3-6 months. The recommended dose of each prescription in the SmPC (Fachinformation“) was compared to the dose that had been actually prescribed.
Results: Out of 232 consecutively patients >65 yrs, 102 had an eGFR <60 ml/min, 16 of these had an eGFR <30 ml/min. The eGFR was closely correlated (r2=0.81) with an independent second eGFR. Out of these 102 patients, 48 had a serum creatinine level within the normal range. Renal adjustment was required in 263 of a total of 613 prescriptions. 72 prescriptions in a total of 45 patients were not appropriately adjusted (32) or prescribed despite a contraindication (40). For chronic prescriptions, metformin, ramipril, enalapril, HCTZ, and spironolactone accounted for 70% of inappropriate dosing; the magnitude of misdosing was 1.5 to 4 fold (median 2). 9 temporary prescriptions (of a total of 60 prescriptions) in 8 patients were not adjusted (cefuroxim, cefpodoxim, levofloxacin). We could not prove that patients with normal serum creatinine had a higher rate of inappropriate dosing than those with already elevated creatinine.
Discussion and conclusion: In this GP practice, we have demonstrated a considerable prevalence of inappropriate dosing in patients with impaired renal function. It remains to be elucidated whether surveillance of appropriate dosing in renal impairment can be optimized e.g. with CPOE.
Recent ecological observations on growth rates and seed production in Isopogon prostratus (Proteaceae), a little-known prostrate shrub from south-eastern NSW and Victoria
Lotte von Richter
- Observations on the longevity and ecology of Isopogon prostratus McGill. (Proteaceae) based on 1985 and 2009 field measures on Newnes Plateau, near Lithgow, and a seed germination trial are provided. Its survival strategy appears to be that of a stress-tolerator with long-term persistence at (relatively few) suitable sites, and it remains a relatively rare plant. It is conjectured that it is likely to have been a species of greater abundance in the drier, colder and generally treeless conditions of the Newnes Plateau 15–20 000 years ago, but, as conditions became warmer and wetter it has become reduced to isolated populations as taller shrubs outcompeted it for light.
Whipcord plants: a comparison of south-eastern Australia with New Zealand
Robert F. Parsons
- Whipcord plant is a term used for some dicot angiosperms with small, scale-like leaves closely appressed to the stem. So far, the term has mostly been used in this sense for plants from New Zealand. Here, I summarize the incidence and habitat relations of New Zealand whipcord plants and then use the literature to show that whipcord plants also occur in south-eastern Australia. New Zealand whipcord plants comprise nine species of Hebe, four of Leonohebe and six of Helichrysum, while in south-eastern Australia there are six species of Ozothamnus and one of Leucophyta. In both areas, some species are alpine to subalpine, while some are from lowland habitats with significant summer water deficits.
Native vegetation of southeast NSW: a revised classification and map for the coast and eastern tablelands
M. G. Tozer
D. A. Keith
- Native vegetation of the NSW south coast, escarpment and southeast tablelands was classified into 191 floristic assemblages at a level of detail appropriate for the discrimination of Threatened Ecological Communities and other vegetation units referred to in government legislation. Assemblages were derived by a numerical analysis of 10832 field sample quadrats including 8523 compiled from 63 previous vegetation surveys. Past bias in the distribution of field data towards land under public tenure was corrected by extensive surveys carried out on private land. The classification revises and integrates the units described in recent vegetation studies of Eden, Cumberland Plain and Sydney-south coast into a single, consistent classification. Relationships between floristic assemblages and climate, terrain, substrate and vegetation structure were used to map the distribution of communities prior to clearing at 1:100 000 scale. The extent of clearing was mapped using interpretations of remote imagery (1991–2001) from previous work, standardised and merged into a single coverage and supplemented with additional work. Profiles for each assemblage, which we term ‘communities’ or ‘map units’, describe their species composition, vegetation structure, environmental habitat, the extent of clearing and conservation status. Lists of diagnostic species were defined using a statistical fidelity measure and a procedure for using these for community identification is described. Approximately 66% of the study area retains a cover of native vegetation, primarily in areas with low fertility soils and dissected topography. Communities subject to over-clearing (>70%) are concentrated in a few large areas characterised by clay/loam soils and flat to undulating terrain. These include the Sydney metropolis, Wingecarribee Plateau, Illawarra Plain, Shoalhaven floodplain, Araluen Valley and Bega Valley, and various smaller river valleys. Forty-one percent of remaining native vegetation is protected within conservation reserves while 31% occurs on private land, 20% in State Forests and 8% on other Crown lands. Forty-five Threatened Ecological Communities (TECs) were recorded in the study area. The majority of TECs are represented by a single map unit, although in some cases a TEC is included within a broader map unit. Twelve TECs are represented by combinations of two or more map units.
Coastal Sandplain Vegetation at Brisbane Water and Broken Bay – reconstructing the past to plan for the future
- The vegetation and floristics of the coastal sandplains on the Umina-Woy Woy Peninsula on the northern foreshores of Broken Bay (lat 33° 30’ S, long 151° 15’ E), 40 km north of Sydney, are described from historical records, sampling of remnants and analysis of regional scale vegetation. Of the seven vegetation communities described, Umina Coastal Sandplain Woodland (UCSW) was originally the most extensive type of vegetation over the Umina-Woy Woy sandplain and on the seaward side of the Pearl Beach sandplain, and possibly on the sandplains at Patonga and Little Patonga. Characteristic tree species are Angophora floribunda and Eucalyptus botryoides; the latter appears to be more common at foreshore sites. Close to the sea and in swales at the base of hillslopes, littoral rainforest elements can be present. Patonga may have had significant inclusions of this vegetation. As a result of clearing for suburban development and its reduction to small remnants, UCSW and Freshwater Wetlands have been listed as an Endangered Ecological Communities under the NSW Threatened Species Conservation Act. A form of the closely-related Sydney Red Gum (SRG) complex occurs on a different SLU on the south-west side of Pearl Beach. A characteristic tree is Angophora costata. Site environmental differences between UCSW and the Sydney Red Gum Complex include their occurrence on iron podsols and humus podsols respectively. Regional vegetation classification and analysis shows that these vegetation units are distinctly different from each other. This is supported by historical evidence from surveyor notation on Crown Survey and land subdivision plans. Originally the Bangalay – Rough-barked Apple Woodland vegetation component of the Umina Woy Woy sandplain was defined by the NSW Scientific Committee for Umina Coastal Sandplain Woodland. Regional analysis now reveals the Red Gum-Red Bloodwood (RGBW) component now merges with this former community. The Pearl Beach vegetation remains separate. A re-definition of UCSW is now required. Management, particularly of UCSW, currently involves revegetation and regeneration works in the vicinity of existing reserves. However, because the depletion has been so extensive there is further opportunity to decrease the loss by utilising the wide riparian reserves and laneways where mature trees still exist. A major conservation concern is the modification and loss of the sandplain vegetation, particularly the wetlands. The historical Crown Survey plans highlight the extent of wetlands as an important ecological feature of the original sandplain landscape. The current study estimated that 83% of wetlands and 79% of riparian vegetation has been lost on the Umina-Woy Woy sandplain since European settlement.
Biomass and floristic patterns in the ground layer vegetation of box-gum grassy eucalypt woodland in Goorooyarroo and Mulligans Flat Nature Reserves, Australian Capital Territory
A. O. Nicholls
Adrian D. Manning
David Bruce Lindenmayer
- We establish a methodology and present baseline data for a long-term grassy woodland restoration study that commenced in 2007 in two nature reserves (Mulligans Flat, Goorooyarroo (35° 9–13’ S; 149° 9–12’ E)) totalling 1386 ha on the northern boundary of Canberra, in the Australian Capital Territory in south eastern Australia. The experimental infrastructure comprises 96 × 1 ha sites established in Eucalyptus blakelyi / Eucalyptus melliodora dominated woodland. These are being subjected to varying kangaroo grazing pressure and augmentation with logs, while burning treatments are planned. One reserve (Mulligans Flat) has been fenced for feral predator control and contains half the sites, forming a companion experiment to Goorooyarroo. Our baseline floristic study comprised estimates, at the site level, of ground layer biomass, species biomass, ground cover types and soil (0–10 cm) properties. From these data we conclude that the groundlayer vegetation is dominated by Joycea pallida, Austrodanthonia spp., Themeda australis and Aristida ramosa. These grasses varied in abundance according to differences in soil pH, phosphorus and to a lesser extent nitrates. Forb frequencies were highly sensitive to nitrate levels with annual exotic forbs dominating at high nitrate sites. More generally, soil nutrient levels and exotic species in some sites indicated areas of previous pasture improvement activities. Biomass estimates indicated extremely high grazing pressure, sufficient to negatively affect the habitat quality for ground-dependent fauna and some soil processes. These data will provide an important basis for examining rates of ecosystem recovery under different restoration strategies.
Waratah theft in Brisbane Water National Park – an analysis of the blue paint poaching reduction program
Catherine A. Offord
- The flowers of Waratahs, Telopea speciosissima (family Proteaceae) are regularly harvested illegally from natural bushland, particularly close to urban areas such as the New South Wales Central Coast. The removal of Waratah blooms from the wild may have implications for the long-term survival of local populations because of the interaction between wildfire events, subsequent flowering and limited seedling recruitment opportunities. To reduce the incidence of theft, blue acrylic paint was applied to blooms to reduce their commercial value. The painting of blooms in 2004 did not significantly reduce the incidence of wildflower theft when compared to unpainted blooms, but overall losses were lower (27%) than in 2003 (33%). However, painting of blooms had a deleterious affect on fruit production on plants with multiple heads with painted blooms having significantly reduced fruit set compared to unpainted blooms. Painting of blooms had no significant effect on seed quality (seed production per fruit, seed germination or seedling vigour) when compared to unpainted blooms. The painting of Waratah blooms to reduce theft was relatively ineffective and decreased fruit production. Alternative strategies should be considered to reduce wildflower theft in the area.
New South Wales Vegetation classification and Assessment: Part 3 Plant communities of the NSW Brigalow Belt South, Nandewar and west New England Bioregions and update of NSW Western Plains and South-western Slopes plant communities. Version 3 of the NSWVCA database
John S. Benson
P. G. Richards
Chris B. Allen
- This fourth paper in the NSW Vegetation Classification and Assessment series covers the Brigalow Belt South-/1(BBS) and Nandewar (NAN) Bioregions and the western half of the New England Bioregion (NET), an area of 9.3 million hectares being 11.6% of NSW. It completes the NSWVCA coverage for the Border Rivers-Gwydir and Namoi CMA areas and records plant communities in the Central West and Hunter–Central Rivers CMA areas. In total, 585 plant communities are now classified in the NSWVCA covering 11.5 of the 18 Bioregions in NSW (78% of the State). Of these 226 communities are in the NSW Western Plains and 416 are in the NSW Western Slopes. 315 plant communities are classified in the BBS, NAN and west-NET Bioregions including 267 new descriptions since Version 2 was published in 2008. Descriptions of the 315 communities are provided in a 919 page report on the DVD accompanying this paper along with updated reports on other inland NSW bioregions and nine Catchment Management Authority areas fully or partly classified in the NSWVCA to date. A read-only version of Version 3 of the NSWVCA database is on the DVD for use on personal computers. A feature of the BBS and NAN Bioregions is the array of ironbark and bloodwood Eucalyptusdominated shrubby woodlands on sandstone and acid volcanic substrates extending from Dubbo to Queensland. This includes iconic natural areas such as Warrumbungle and Mount Kaputar National Parks and the 500,000 ha Pilliga Scrub forests. Large expanses of basalt-derived soils support grassy box woodland and native grasslands including those on the Liverpool Plains; near Moree; and around Inverell, most of which are cleared and threatened. Wetlands occur on sodic soils near Yetman and in large clay gilgais in the Pilliga region. Sedgelands are rare but occupy impeded creeks. Aeolian lunettes occur at Narran Lake and near Gilgandra. Areas of deep sand contain Allocasuarina, eucalypt mallee and Melaleuca uncinata heath. Tall grassy or ferny open forests occur on mountain ranges above 1000m elevation in the New England Bioregion and on the Liverpool Range while grassy box woodlands occupy lower elevations with lower rainfall and higher temperatures. The vegetation classification and assessment is based on over 100 published and unpublished vegetation surveys and map unit descriptions, expert advice, extra plot sampling and data analysis and over 25 000 km of road traverse with field checking at 805 sites. Key sources of data included floristic analyses produced in western regional forest assessments in the BBS and NAN Bioregions, floristic analyses in over 60 surveys of conservation reserves and analysis of plot data in the western NET Bioregion and covering parts of the Namoi and Border Rivers- Gwydir CMA areas. Approximately 60% of the woody native vegetation in the study area has been cleared resulting in large areas of “derived” native grasslands. As of June 2010, 7% of the area was in 136 protected areas and 127 of the 315 plant communities were assessed to be adequately protected in reserves. Using the NSWVCA database threat criteria, 15 plant communities were assessed as being Critically Endangered, 59 Endangered, 60 Vulnerable, 99 Near Threatened and 82 Least Concern. 61 of these communities are assessed as part of NSW or Commonwealth-listed Threatened Ecological Communities. Current threats include expanding dryland and irrigated cropping on alluvial plains, floodplains and gently undulating topography at lower elevations; over-grazing of steep hills; altered water tables and flooding regimes; localized mining; and the spread of exotic species, notably Coolatai Grass (Hyparrhenia hirta).