Role of vascular endothelial growth factor and placental growth factors during retinal vascular development and hyaloid regression

PURPOSE. Vascular endothelial growth factor (VEGF)-A and placental growth factor (PIGF) are members of a large group of homologous peptides identified as the VEGF family. Although VEGF-A is known to act as a potent angiogenic peptide in the retina, the vasoactive function of PIGF in this tissue is less well defined. This study has sought to elucidate the expression patterns and modulatory role of these growth factors during retinal vascular development and hyaloid regression in the neonatal mouse. METHODS. C57BL6J mice were killed at postnatal days (P)1, P3, P5, P7, P9, and P11. The eyes were enucleated and processed for in situ hybridization and immunocytochemistry and the retinas extracted for total protein or RNA. Separate groups of neonatal mice were also injected intraperitoneally daily from P2 through P9 with either VEGF-neutralizing antibody, PIGF-neutralizing antibody, isotype immunoglobulin (Ig)-G, or phosphate-buffered saline (PBS). The mice were then perfused with fluorescein isothiocyanate (FITC)-dextran, and the eyes were subsequently embedded in paraffin wax or flat mounted. RESULTS. Quantitative (real-time) reverse transcription-polymerase chain reaction (RT-PCR) demonstrated similar expression patterns of VEGF-A and PIGF mRNA during neonatal retinal development, although the fluctuation between time periods was greater overall for VEGF-A. The localization of VEGF-A and PIGF in the retina, as revealed by in situ hybridization and immunohistochemistry, was also similar. Neutralization of VEGF-A caused a significant reduction in the hyaloid and retinal vasculature, whereas PIGF antibody treatment caused a marked persistence of the hyaloid without significantly affecting retinal vascular development. CONCLUSIONS. Although having similar expression patterns in the retina, these growth factors appear to have distinct modulatory influences during normal retinal vascular development and hyaloid regression.

Toxicity of mildly modified low-density lipoproteins to cultured retinal capillary endothelial cells and pericytes

To investigate the role of modified low-density lipoproteins (LDL) in the pathogenesis of diabetic retinopathy, we studied the cytotoxicity of normal and mildly modified human LDL to bovine retinal capillary endothelial cells and pericytes in vitro. Pooled LDL was incubated (in phosphate-buffered saline-EDTA, 3 days, 37 degrees C) under 1) nitrogen with additional chelating agents and 2) air, to prepare normal and minimally oxidized LDL, respectively. Similar conditions, but with the addition of 50 mM D-glucose, were used to prepare glycated and glycoxidized LDL. None of the LDL preparations was recognized by the macrophage scavenger receptor, confirming limited modification. Retinal capillary endothelial cells and pericytes were grown to confluence and then exposed for 2 or 3 days to serum-free medium (1% albumin) supplemented with normal or modified LDL (100 mg/l) or to serum-free medium alone. Cytotoxicity was assessed by cell counting (live and total cells) and by cell protein determination. Compared with normal LDL, modified LDL were cytotoxic to both cell types at both time points, causing highly significant decreases in live and total cell counts (P <0.001) (analysis of variance). Reductions in cell protein also were significant for pericytes at day 3 (P = 0.016) and of borderline significance for endothelial cells at day 2 (P = 0.05) and day 3 (P = 0.063). Cytotoxicity increased as follows: normal <glycated <or = minimally oxidized <glycoxidized LDL. We conclude that, in diabetes, mild modification of LDL resulting from separate or combined processes of glycation and oxidation may contribute to chronic retinal capillary injury and thus to the development of diabetic retinopathy.

Coleoptera and microbe biomass in Antarctic Dry Valley paleosols adjacent to the Inland Ice: Implications for Mars

Bulk paleosol samples collected from a Middle to Early Miocene moraine in the New Mountain area of the Dry Valleys, Antarctica, yielded Coleoptera exoskeletons and occasional endoskeletons showing considerable diagenetic effects along with several species of bacteria, all lodged in a dry-frozen but salt-rich horizon at shallow depth to the land surface. The till is at the older end of a chronologic sequence of glacial deposits, thought to have been deposited before the transition from wet-based to cold-based ice (similar to 15 Ma), and hence, entirely weathered in contact with the subaerial atmosphere. It is possible, though not absolutely verifiable, that the skeletons date from this early stage of emplacement having undergone modifications whenever light snowmelt occurred or salt concentrations lowered the freezing temperature to maintain water as liquid. Correlation of the Coleoptera species with cultured bacteria in the sample and the likelihood of co-habitation with Beauveria bassiani found in two adjacent, although younger paleosols, leads to new questions about the antiquity of the Coleoptera and the source of N and glucose from chitinase derived from the insects. The skeletons in the 831 section may date close to the oldest preserved chitin (Oligocene) yet found on Earth. While harsh Martian conditions make it seemingly intolerable for complex, multicellular organisms such as insects to exist in the near-surface and subaerially, life within similar cold, dry paleosol microenvironments (Cryosols) of Antarctica point to life potential for the Red Planet, especially when considering the relatively diverse microbe (bacteria and fungi) population. (C) 2011 Elsevier Ltd. All rights reserved.

Later Pleistocene evolution of the Exe valley: A chronstratigraphic model of terrace formation and its implications for Palaeolithic archaeology

This paper presents the ?rst systematic chronostratigraphic study of the river terraces of the Exe catchment in South West England and a new conceptual model for terrace formation in unglaciated basins with applicability to terrace staircase sequences elsewhere. The Exe catchment lay beyond the maximum extent of Pleistocene ice sheets and the drainage pattern evolved from the Tertiary to the Middle Pleistocene, by which time the major valley systems were in place and downcutting began to create a staircase of strath terraces. The higher terraces (8-6) typically exhibit altitudinal overlap or appear to be draped over the landscape, whilst the middle terraces show greater altitudinal separation and the lowest terraces are of a cut and ?ll form. The terrace deposits investigated in this study were deposited in cold phases of the glacial-interglacial Milankovitch climatic cycles with the lowest four being deposited in the Devensian Marine Isotope Stages (MIS) 4-2. A new cascade process-response model is proposed of basin terrace evolution in the Exe valley, which emphasises the role of lateral erosion in the creation of strath terraces and the reworking of inherited resistant lithological components down through the staircase. The resultant emergent valley topography and the reworking of artefacts along with gravel clasts, have important implications for the dating of hominin presence and the local landscapes they inhabited. Whilst the terrace chronology suggested here is still not as detailed as that for the Thames or the Solent System it does indicate a Middle Palaeolithic hominin presence in the region, probably prior to the late Wolstonian Complex or MIS 6. This supports existing data from cave sites in South West England.

Design of an intravaginal ring for the controlled delivery of 17b-estradiol as its 3-acetate ester

Suitable ester prodrugs of 17b-estradiol are identified, thus permitting effective sustained and controlled estrogen replacement therapy (ERT) from an elastomeric, silicone intravaginal ring (IVR). IVR devices of reservoir design were prepared by blending silicone elastomer base with n-propylorthosilicate (cross-linker) and 10% w/w of 17b-estradiol or an ester prodrug, the mix being activated with 0.5% w/w stannous octoate and cured at 808C for 2 min. A rate-controlling membrane was similarly prepared, without the active agent. IVR devices were of cross-sectional diameter 9 mm, outer diameter 54 mm, with core cross-sectional diameter of 2 mm and core length varied as required. Sink conditions were evident for the 17b-estradiol esters in 1.0% aqueous benzalkonium chloride solution. The low release rates into 0.9% w/v saline of the lipophilic valerate and benzoate esters were due to their intrinsically low aqueous solubilities. In vivo, these esters failed to raise plasma estradiol above baseline levels in postmenopausal human volunteers, despite good in vitro release characteristics under sink conditions. The best release rates under sink conditions, in combination with substantial aqueous solubilities as indicated by the release rates into saline, were observed for the acetate and propionate esters. A
combination of drug release characteristics, short plasma half-life and a toxicologically acceptable hydrolysis product indicated that 17b-estradiol-3-acetate was the prodrug of choice for IVR delivery of ERT. In vivo, an IVR device releasing
100 mg/day of estradiol as its 3-acetate ester maintained over 84 days a circulating plasma concentration in the region of 300 pmol l , within the clinically desirable range for ERT.

Multi-Objective Reactive Power Support from Wind farms for Network Performance Enhancement

This paper examines the ability of the doubly fed induction generator (DFIG) to deliver multiple reactive power objectives during variable wind conditions. The reactive power requirement is decomposed based on various control objectives (e.g. power factor control, voltage control, loss minimisation, and flicker mitigation) defined around different time frames (i.e. seconds, minutes, and hourly), and the control reference is generated by aggregating the individual reactive power requirement for each control strategy. A novel coordinated controller is implemented for the rotor-side converter and the grid-side converter considering their capability curves and illustrating that it can effectively utilise the aggregated DFIG reactive power capability for system performance enhancement. The performance of the multi-objective strategy is examined for a range of wind and network conditions, and it is shown that for the majority of the scenarios, more than 92% of the main control objective can be achieved while introducing the integrated flicker control scheme with the main reactive power control scheme. Therefore, optimal control coordination across the different control strategies can maximise the availability of ancillary services from DFIG-based wind farms without additional dynamic reactive power devices being installed in power networks.

Active use of DFIG based Wind Farms for Transient Stability Improvement during Grid Disturbances

The doubly-fed induction generator (DFIG) now represents the dominant technology in wind turbine design. One consequence of this is limited damping and inertial response during transient grid disturbances. A dasiadecoupledpsila strategy is therefore proposed to operate the DFIG grid-side converter (GSC) as a static synchronous compensator (STATCOM) during a fault, supporting the local voltage, while the DFIG operates as a fixed-speed induction generator (FSIG) providing an inertial response. The modeling aspects of the decoupled control strategy, the selection of protection control settings, the significance of the fault location and operation at sub- and super-synchronous speeds are analyzed in detail. In addition, a case study is developed to validate the proposed strategy under different wind penetrations levels. The simulations show that suitable configuration of the decoupled strategy can be deployed to improve system voltage stability and inertial response for a range of scenarios, especially at high wind penetration. The conclusions are placed in context of the practical limitations of the technology employed and the system conditions.

Towards groundwater security in coastal East Africa: initiating a regional research network and integrated hydrogeologic, climatic & socio-economic observatories in coastal aquifers of the Comoros Islands, Kenya and Tanzania

African coastal regions are expected to experience the highest rates of population growth in coming decades. Fresh groundwater resources in the coastal zone of East Africa (EA) are highly vulnerable to seawater intrusion. Increasing water demand is leading to unsustainable and ill-planned well drilling and abstraction. Wells supplying domestic, industrial and agricultural needs are or have become, in many areas, too saline for use. Climate change, including weather changes and sea level rise, is expected to exacerbate this problem. The multiplicity of physical, demographic and socio-economic driving factors makes this a very challenging issue for management. At present the state and probable evolution of coastal aquifers in EA are not well documented. The UPGro project 'Towards groundwater security in coastal East Africa' brings together teams from Kenya, Tanzania, Comoros Islands and Europe to address this knowledge gap. An integrative multidisciplinary approach, combining the expertise of hydrogeologists, hydrologists and social scientists, is investigating selected sites along the coastal zone in each country. Hydrogeologic observatories have been established in different geologic and climatic settings representative of the coastal EA region, where focussed research will identify the current status of groundwater and identify future threats based on projected demographic and climate change scenarios. Researchers are also engaging with end users as well as local community and stakeholder groups in each area in order to understanding the issues most affecting the communities and searching sustainable strategies for addressing these.

Integrating geophysical data and upscaling techniques to support regional groundwater flow modelling: A practical example of the Lagan Valley, Northern Ireland

When studying heterogeneous aquifer systems, especially at regional scale, a degree of generalization is anticipated. This can be due to sparse sampling regimes, complex depositional environments or lack of accessibility to measure the subsurface. This can lead to an inaccurate conceptualization which can be detrimental when applied to groundwater flow models. It is important that numerical models are based on observed and accurate geological information and do not rely on the distribution of artificial aquifer properties. This can still be problematic as data will be modelled at a different scale to which it was collected. It is proposed here that integrating geophysics and upscaling techniques can assist in a more realistic and deterministic groundwater flow model. In this study, the sedimentary aquifer of the Lagan Valley in Northern Ireland is chosen due to intruding sub-vertical dolerite dykes. These dykes are of a lower permeability than the sandstone aquifer. The use of airborne magnetics allows the delineation of heterogeneities, confirmed by field analysis. Permeability measured at the field scale is then upscaled to different levels using a correlation with the geophysical data, creating equivalent parameters that can be directly imported into numerical groundwater flow models. These parameters include directional equivalent permeabilities and anisotropy. Several stages of upscaling are modelled in finite element. Initial modelling is providing promising results, especially at the intermediate scale, suggesting an accurate distribution of aquifer properties. This deterministic based methodology is being expanded to include stochastic methods of obtaining heterogeneity location based on airborne geophysical data. This is through the Direct Sample method of Multiple-Point Statistics (MPS). This method uses the magnetics as a training image to computationally determine a probabilistic occurrence of heterogeneity. There is also a need to apply the method to alternate geological contexts where the heterogeneity is of a higher permeability than the host rock.

North Atlantic marine radiocarbon reservoir ages through Heinrich event H4: A new method for marine age model construction

Cooling and sinking of dense saline water in the Norwegian–Greenland Sea is essential for the formation of North Atlantic Deep Water. The convection in the Norwegian–Greenland Sea allows for a northward flow of warm surface water and southward transport of cold saline water. This circulation system is highly sensitive to climate change and has been shown to operate in different modes. In ice cores the last glacial period is characterized by millennial-scale Dansgaard–Oeschger (D–O) events of warm interstadials and cold stadials. Similar millennial-scale variability (linked to D–O events) is evident from oceanic cores, suggesting a strong coupling of the atmospheric and oceanic circulations system. Particularly long-lasting cold stadials correlate with North Atlantic Heinrich events, where icebergs released from the continents caused a spread of meltwater over the northern North Atlantic and Nordic seas. The meltwater layer is believed to have caused a stop or near-stop in the deep convection, leading to cold climate. The spreading of meltwater and changes in oceanic circulation have a large influence on the carbon exchange between atmosphere and the deep ocean and lead to profound changes in the 14C activity of the surface ocean. Here we demonstrate marine 14C reservoir ages (R) of up to c. 2000 years for Heinrich event H4. Our R estimates are based on a new method for age model construction using identified tephra layers and tie-points based on abrupt interstadial warmings.

Microabrasion-corrosion of cast CoCrMo alloy in simulated body fluids

Wear and corrosion of metal-on-metal hip replacements results in wear debris and metal-ion release in vivo, which may subsequently cause pain and hypersensitivity for patients. Retrieved metal-on-metal hip replacements have revealed that two-body sliding wear and three-body abrasive wear are the predominant wear mechanisms. However, there is a lack of understanding of the combined effects of wear/corrosion, especially the effect of abrasion-corrosion.

This study investigates the sliding-corrosion and abrasion-corrosion performance of a cast CoCrMo alloy in simulated hip joint environments using a microabrasion rig integrated with an electrochemical cell. Tests have been conducted in 0.9% NaCl, phosphate buffered saline solution, 25% and 50% bovine serum solutions with 0 or 1 g cm(-3) SiC at 37 degrees C. Experimental results reveal that under abrasion-corrosion test conditions, the presence of proteins increased the total specific wear rate. Conversely, electrochemical noise measurements indicated that the average anodic current levels were appreciably lower for the proteinaceous solutions when compared with the inorganic solutions. A severely deformed nanocrystalline layer was identified immediately below the worn surface for both proteinaceous and inorganic solutions. The layer is formed by a recrystallisation process and/or a strain-induced phase transformation that occurs during microabrasion-corrosion. (C) 2008 Elsevier Ltd. All rights reserved.

Is there a common water-activity limit for the three domains of life?

Archaea and Bacteria constitute a majority of life systems on Earth but have long been considered inferior to Eukarya in terms of solute tolerance. Whereas the most halophilic prokaryotes are known for an ability to multiply at saturated NaCl (water activity (a_w) 0.755) some xerophilic fungi can germinate, usually at high-sugar concentrations, at values as low as 0.650–0.605 a_w. Here, we present evidence that halophilic prokayotes can grow down to water activities of <0.755 for Halanaerobium lacusrosei (0.748), Halobacterium strain 004.1 (0.728), Halobacterium sp. NRC-1 and Halococcus morrhuae (0.717), Haloquadratum walsbyi (0.709), Halococcus salifodinae (0.693), Halobacterium noricense (0.687), Natrinema pallidum (0.681) and haloarchaeal strains GN-2 and GN-5 (0.635 a_w). Furthermore, extrapolation of growth curves (prone to giving conservative estimates) indicated theoretical minima down to 0.611 a_w for extreme, obligately halophilic Archaea and Bacteria. These were compared with minima for the most solute-tolerant Bacteria in high-sugar (or other non-saline) media (Mycobacterium spp., Tetragenococcus halophilus, Saccharibacter floricola, Staphylococcus aureus and so on) and eukaryotic microbes in saline (Wallemia spp., Basipetospora halophila, Dunaliella spp. and so on) and high-sugar substrates (for example, Xeromyces bisporus, Zygosaccharomyces rouxii, Aspergillus and Eurotium spp.). We also manipulated the balance of chaotropic and kosmotropic stressors for the extreme, xerophilic fungi Aspergillus penicilloides and X. bisporus and, via this approach, their established water-activity limits for mycelial growth (~0.65) were reduced to 0.640. Furthermore, extrapolations indicated theoretical limits of 0.632 and 0.636 a_w for A. penicilloides and X. bisporus, respectively. Collectively, these findings suggest that there is a common water-activity limit that is determined by physicochemical constraints for the three domains of life.

THE CULTURE MICROBIOME IN THE LUNGS OF PATIENTS WITH COPD

Introduction and Aims: Previous studies have shown that the lungs of Cystic Fibrosis (CF) and bronchiectasis (BE, not caused by CF) patients are colonised by a range of aerobic and anaerobic bacteria. As bacteria are also implicated in the pathogenesis and progression of chronic obstructive pulmonary disease (COPD), this study aimed to determine the culture microbiome of the COPD airways.

Methods: Samples were collected from 13 stable COPD patients during routine bronchoscopy. Bronchial washings were taken at a single location in the right middle lobe by flushing and removing 30 ml of sterile saline. Samples were cultured under strict anaerobic conditions with bacteria detected by plating on both selective and non-selective agar media and quantified by total viable count (TVC). Identification of the cultured bacteria was performed by amplification and subsequent sequencing of the 16sRNA gene.

Results: Mean FEV1 was 1.36 (range 0.84–2.26, mean per cent predicted FEV1, 54%), and the mean ratio (FEV1/FVC) was 51%. Bacteria were detected in 12/13 samples (92%) with bacteria from the genera Streptococcus [12/13 samples, 92%; mean (range) TVC 9.62×105 cfu/ml (1.50×103–1.42×107)] and Haemophilus [4/13 samples, 31%; mean (range) 6.40×104 cfu/ml (2.20×103–1.60×105)] most frequently detected. Anaerobic bacteria primarily from the genera Prevotella [8/13 samples, 62%; mean (range) TVC 1.12×104 cfu/ml (1.30×103–4.20×104)] and Veillonella [5/13 samples, 38%; mean (range) TVC 1.29×105 cfu/ml (4.20×103–3.60×105)] were also detected. Pseudomonas and Moraxella were not detected in any samples.

Conclusions: Our results show that bacteria from the genera Streptococcus, Haemophilus, Prevotella and Veillonella are frequently present the airways of patients suffering from COPD. Taking account of the dilutional effect of the bronchial wash procedure and extrapolating to allow comparison with sputum data in our laboratory for CF and BE, the relative load of bacteria from the genera Streptococcus, Prevotella and Veillonella is similar in these three airway diseases. The potential role of these bacteria in the progression and pathogenesis of COPD requires further investigation.