Retargeting of pre-set regions on chromosome for high gene expression in mammalian cells

We have developed a system to hunt and reuse special gene integration sites that allow for high and stable gene expression. A vector, named pRGFP8, was constructed. The plasmid pRGFP8 contains a reporter gene, gfp2 and two extraneous DNA fragments. The gene gfp2 makes it possible to screen the high expression regions on the chromosome. The extraneous DNA fragments can help to create the unique loci on the chromosome and increase the gene targeting frequency by increasing the homology. After transfection into Chinese hamster ovary cells (CHO) cells, the linearized pRGFP8 can integrate into the chromosome of the host cells and form the unique sites. With FACS, 90 millions transfected cells were sorted and the cells with strongest GFP expression were isolated, and then 8 stable high expression GFP CHO cell lines were selected as candidates for the new host cell. Taking the unique site created by pRGFP8 on the chromosome in the new host cells as a targeting locus, the gfp2 gene was replaced with the gene of interest, human ifngamma, by transfecting the targeting plasmid pRIH-IFN. Then using FACS, the cells with the dimmest GFP fluorescence were selected. These cells showed they had strong abilities to produce the protein of interest, IFN-gamma. During the gene targeting experiment, we found there is positive correlation between the fluorescence density of the GFP CHO host cells and the specific production rate of IFN-gamma. This result shows that the strategy in our expression system is correct: the production of the interesting protein increases with the increase fluorescence of the GFP host cells. This system, the new host cell lines and the targeting vector, can be utilized for highly expressing the gene of interest. More importantly, by using FACS, we can fully screen all the transfected cells, which can reduce the chances of losing the best cells.

Análisis de la influencia de los organismos adscritos a la organización de naciones unidas (UNICEF- ECPAT) en la solución de la problemática de la trata de niños en el sudeste asiático continental. periodo 2000-2010

La trata de niños es un fenómeno cada vez más relevante en el sistema internacional y ha generado importantes esfuerzos de cooperación de agencias internacionales como UNICEF y ECPAT. En ese sentido, ambos organismos han enfocado su actividad en las regiones donde se han registrado los mayores índices de violación de los Derechos Humanos. Este Estudio de Caso se ha centrado en el Sudeste Asiático continental conformado por Tailandia, Laos, Camboya, Vietnam, Singapur, Myanmar y Malasia; con el fin de determinar de qué manera han trabajado ambos organismos en la región y de qué forma han contribuido a la solución de esta problemática por medio de la firma de acuerdos internacionales, implementación de legislación nacional y programas de cooperación. A pesar del trabajo realizado no solo por entidades internacionales sino también por autoridades gubernamentales, no se ha observado una disminución real de las cifras.

Linking Canadian Harvested Juvenile American Black Ducks to Their Natal Areas Using Stable Isotope (δD, δ13C, and δ15N) Methods

Understanding source-sink dynamics of game birds is essential to harvest and habitat management but acquiring this information is often logistically and financially challenging using traditional methods of population surveys and banding studies. This is especially true for species such as the American Black Duck (Anas rubripes), which have low breeding densities and extensive breeding ranges that necessitate extensive surveys and banding programs across eastern North America. Despite this effort, the contribution of birds fledged from various landscapes and habitat types within specific breeding ranges to regional harvest is largely unknown but remains an important consideration in adaptive harvest management and targeted habitat conservation strategies. We investigated if stable isotope (δD, δ13C, δ15N) could augment our present understanding of connectivity between breeding and harvest areas and so provide information relevant to the two main management strategies for black ducks, harvest and habitat management. We obtained specimens from 200 hatch-year Black Duck wings submitted to the Canadian Wildlife Service Species Composition Survey. Samples were obtained from birds harvested in Western, Central, and Eastern breeding/harvest subregions to provide a sample representative of the range and harvest rate of birds harvested in Canada. We sampled only hatch-year birds to provide an unambiguous and direct link between production and harvest areas. Marine origins were assigned to 12%, 7%, and 5% of birds harvested in the Eastern, Central, and Western subregions, respectively. In contrast, 32%, 9%, and 5% of birds were assigned, respectively, to agricultural origins. All remaining birds were assigned to nonagricultural origins. We portrayed probability of origin using a combination of Bayesian statistical and GIS methods. Placement of most eastern birds was western Nova Scotia, eastern New Brunswick, Prince Edward Island, and southern Newfoundland. Agricultural birds from the Central region were consistent with the Saguenay region of Québec and the eastern claybelt with nonagricultural birds originating in the boreal. Western nonagricultural birds were associated with broad boreal origins from southern James Bay to Lake of the Woods and east to Cochrane, Ontario. Our work shows that the geographic origins, landscape, and habitat associations of hatch-year Black Ducks can be inferred using this technique and we recommend that a broad-scale isotopic study using a large sample of Canadian and US harvested birds be implemented to provide a continental perspective of source-sink population dynamics.

Chemical composition observed over the mid-Atlantic and the detection of pollution signatures far from source regions

The atmospheric composition of the central North Atlantic region has been sampled using the FAAM BAe146 instrumented aircraft during the Intercontinental Transport of Ozone and Precursors (ITOP) campaign, part of the wider International Consortium for Atmospheric Research on Transport and Transformation (ICARTT). This paper presents an overview of the ITOP campaign. Between late July and early August 2004, twelve flights comprising 72 hours of measurement were made in a region from approximately 20 to 40°W and 33 to 47°N centered on Faial Island, Azores, ranging in altitude from 50 to 9000 m. The vertical profiles of O3 and CO are consistent with previous observations made in this region during 1997 and our knowledge of the seasonal cycles within the region. A cluster analysis technique is used to partition the data set into air mass types with distinct chemical signatures. Six clusters provide a suitable balance between cluster generality and specificity. The clusters are labeled as biomass burning, low level outflow, upper level outflow, moist lower troposphere, marine and upper troposphere. During this summer, boreal forest fire emissions from Alaska and northern Canada were found to provide a major perturbation of tropospheric composition in CO, PAN, organic compounds and aerosol. Anthropogenic influenced air from the continental boundary layer of the USA was clearly observed running above the marine boundary layer right across the mid-Atlantic, retaining high pollution levels in VOCs and sulfate aerosol. Upper level outflow events were found to have far lower sulfate aerosol, resulting from washout on ascent, but much higher PAN associated with the colder temperatures. Lagrangian links with flights of other aircraft over the USA and Europe show that such signatures are maintained many days downwind of emission regions. Some other features of the data set are highlighted, including the strong perturbations to many VOCs and OVOCs in this remote region.

Gene expression profiling of human hibernating myocardium: Increased expression of B-type natriuretic peptide and proenkephalin in hypocontractile vs normally-contracting regions of the heart

A greater understanding of the molecular basis of hibernating myocardium may assist in identifying those patients who would most benefit from revascularization. Paired heart biopsies were taken from hypocontractile and normally-contracting myocardium (identified by cardiovascular magnetic resonance) from 6 patients with chronic stable angina scheduled for bypass grafting. Gene expression profiles of hypocontractile and normally-contracting samples were compared using Affymetrix microarrays. The data for patients with confirmed hibernating myocardium were analysed separately and a different, though overlapping, set (up to 380) of genes was identified which may constitute a molecular fingerprint for hibernating myocardium. The expression of B-type natriuretic peptide (BNP) was increased in hypocontractile relative to normally-contracting myocardium. The expression of BNP correlated most closely with the expression of proenkephalin and follistatin 3, which may constitute additional heart failure markers. Our data illustrate differential gene expression in hypocontractile and/hibernating myocardium relative to normally-contracting myocardium within individual human hearts. Changes in expression of these genes, including increased relative expression of natriuretic and other factors, may constitute a molecular signature for hypocontractile and/or hibernating myocardium.

The core-periphery model with three regions and more

We determine the properties of the core-periphery model with three regions and compare our results with those of the standard 2-region model. The conditions for the stability of dispersion and concentration are established. As in the 2-region model, dispersion and concentration can be simultaneously stable. We show that the 3-region (2-region) model favours the concentration (dispersion) of economic activity. Furthermore, we provide some results for the n-region model. We show that the stability of concentration of the 2-region model implies that of any model with an even number of regions.

Exploiting a hybrid environmental design strategy in the continental climate of Beijing

The built environment in China is required to achieve a 50% reduction in carbon emissions by 2020 against the 1980 design standard. A particular challenge is how to maintain acceptable comfort conditions through the hot humid summers and cold desiccating winters of its continental climate regions. Fully air-conditioned sealed envelopes, often fully glazed, are becoming increasingly common in these regions. Remedial strategies involve technical refinements to the air-handling equipment and a contribution from renewable energy sources in an attempt to achieve the prescribed net reduction in energy use. However an alternative hybrid environmental design strategy is developed in this research project. It exploits observed temperate periods of weeks, days, even hours in duration to free-run an office and exhibition building configured to promote natural stack ventilation when ambient conditions permit and mechanical ventilation when conditions require it, the two modes delivered through the same physical infrastructure. The proposal is modelled in proprietary software and the methodology adopted is described. The challenge is compounded by its first practical application to an existing reinforced concrete frame originally designed to receive a highly glazed envelope. This original scheme is reviewed in comparison. Furthermore the practical delivery of the proposal value engineered out a proportion of the ventilation stacks. The likely consequence of this for the environmental performance of the building is investigated through a sensitivity study.

Stable isotope and modelling evidence for CO2 as a driver of glacial–interglacial vegetation shifts in southern Africa

Atmospheric CO2 concentration is hypothesized to influence vegetation distribution via tree–grass competition, with higher CO2 concentrations favouring trees. The stable carbon isotope (δ13C) signature of vegetation is influenced by the relative importance of C4 plants (including most tropical grasses) and C3 plants (including nearly all trees), and the degree of stomatal closure – a response to aridity – in C3 plants. Compound-specific δ13C analyses of leaf-wax biomarkers in sediment cores of an offshore South Atlantic transect are used here as a record of vegetation changes in subequatorial Africa. These data suggest a large increase in C3 relative to C4 plant dominance after the Last Glacial Maximum. Using a process-based biogeography model that explicitly simulates 13C discrimination, it is shown that precipitation and temperature changes cannot explain the observed shift in δ13C values. The physiological effect of increasing CO2 concentration is decisive, altering the C3/C4 balance and bringing the simulated and observed δ13C values into line. It is concluded that CO2 concentration itself was a key agent of vegetation change in tropical southern Africa during the last glacial–interglacial transition. Two additional inferences follow. First, long-term variations in terrestrial δ13Cvalues are not simply a proxy for regional rainfall, as has sometimes been assumed. Although precipitation and temperature changes have had major effects on vegetation in many regions of the world during the period between the Last Glacial Maximum and recent times, CO2 effects must also be taken into account, especially when reconstructing changes in climate between glacial and interglacial states. Second, rising CO2 concentration today is likely to be influencing tree–grass competition in a similar way, and thus contributing to the "woody thickening" observed in savannas worldwide. This second inference points to the importance of experiments to determine how vegetation composition in savannas is likely to be influenced by the continuing rise of CO2 concentration.

Meteorological factors controlling low-level continental pollutant outflow across a coast

Coastal outflow describes the horizontal advection of pollutants from the continental boundary layer across a coastline into a layer above the marine boundary layer. This process can ventilate polluted continental boundary layers and thus regulate air quality in highly populated coastal regions. This paper investigates the factors controlling coastal outflow and quantifies its importance as a ventilation mechanism. Tracers in the Met Office Unified Model (MetUM) are used to examine the magnitude and variability of coastal outflow over the eastern United States for a 4 week period during summer 2004. Over the 4 week period, ventilation of tracer from the continental boundary layer via coastal outflow occurs with the same magnitude as vertical ventilation via convection and advection. The relative importance of tracer decay rate, cross-coastal advection rate, and a parameter based on the relative continental and marine boundary layer heights, on coastal outflow is assessed by reducing the problem to a time-dependent box-model. The ratio of the advection rate and decay rate is a dimensionless parameter which determines whether tracers are long-lived or short-lived. Long- and short-lived tracers exhibit different behaviours with respect to coastal outflow. For short-lived tracers, increasing the advection rate increases the diurnally averaged magnitude of coastal outflow, but has the opposite effect for very long-lived tracers. Short-lived tracers exhibit large diurnal variability in coastal outflow but long-lived tracers do not. By combining the MetUM and box-model simulations a landwidth is determined which represents the distance inland over which emissions contribute significantly to coastal outflow. A landwidth of between 100 and 400 km is found to be representative for a tracer with a lifetime of 24 h.

Late Quaternary environmental change in the interior South American tropics: new insight from leaf wax stable isotopes

Stable isotope analysis of leaf waxes in a sediment core from Laguna La Gaiba, a shallow lake located at the Bolivian margin of the Pantanal wetlands, provides new perspective on vegetation and climate change in the lowland interior tropics of South America over the past 40,000 years. The carbon isotopic compositions (δ13C) of long-chain n-alkanes reveal large shifts between C3-and C4-dominated vegetation communities since the last glacial period, consistent with landscape reconstructions generated with pollen data from the same sediment core. Leaf wax δ13C values during the last glacial period reflect an open landscape composed of C4grasses and C3herbs from 41–20ka. A peak in C4abundance during the Last Glacial Maximum (LGM, ∼21ka) suggests drier or more seasonal conditions relative to the earlier glacial period, while the development of a C3-dominated forest community after 20 ka points to increased humidity during the last deglaciation. Within the Holocene, large changes in the abundance of C4 vegetation indicate a transition from drier or more seasonal conditions during the early/mid-Holocene to wetter conditions in the late Holocene coincident with increasing austral summer insolation. Strong negative correlations between leaf wax δ13C and δD values over the entire record indicate that the majority of variability in leaf wax δD at this site can be explained by variability in the magnitude of biosynthetic fractionation by different vegetation types rather than changes in meteoric water δD signatures. However, positive δD deviations from the observed δ13C–δD trends are consistent with more enriched source water and drier or more seasonal conditions during the early/mid-Holocene and LGM. Overall, our record adds to evidence of varying influence of glacial boundary conditions and orbital forcing on South American Summer Monsoon precipitation in different regions of the South American tropics. Moreover, the relationships between leaf wax stable isotopes and pollen data observed at this site underscore the complementary nature of pollen and leaf wax δ13C data for reconstructing past vegetation changes and the potentially large effects of such changes on leaf wax δD signatures.

Selective environmental stress caused by magmatic sulfur emissions from continental flood basalt eruptions

Several biotic crises during the past 300 million years have been linked to episodes of continental flood basalt volcanism, and in particular to the release of massive quantities of magmatic sulphur gas species. Flood basalt provinces were typically formed by numerous individual eruptions, each lasting years to decades. However, the environmental impact of these eruptions may have been limited by the occurrence of quiescent periods that lasted hundreds to thousands of years. Here we use a global aerosol model to quantify the sulphur-induced environmental effects of individual, decade-long flood basalt eruptions representative of the Columbia River Basalt Group, 16.5–14.5 million years ago, and the Deccan Traps, 65 million years ago. For a decade-long eruption of Deccan scale, we calculate a decadal-mean reduction in global surface temperature of 4.5 K, which would recover within 50 years after an eruption ceased unless climate feedbacks were very different in deep-time climates. Acid mists and fogs could have caused immediate damage to vegetation in some regions, but acid-sensitive land and marine ecosystems were well-buffered against volcanic sulphur deposition effects even during century-long eruptions. We conclude that magmatic sulphur from flood basalt eruptions would have caused a biotic crisis only if eruption frequencies and lava discharge rates had been high and sustained for several centuries at a time.

Upper-mantle seismic anisotropy from SKS splitting in the South American stable platform: A test of asthenospheric flow models beneath the lithosphere

Upper-mantle seismic anisotropy has been extensively used to infer both present and past deformation processes at lithospheric and asthenospheric depths. Analysis of shear-wave splitting (mainly from core-refracted SKS phases) provides information regarding upper-mantle anisotropy. We present average measurements of fast-polarization directions at 21 new sites in poorly sampled regions of intra-plate South America, such as northern and northeastern Brazil. Despite sparse data coverage for the South American stable platform, consistent orientations are observed over hundreds of kilometers. Over most of the continent, the fast-polarization direction tends to be close to the absolute plate motion direction given by the hotspot reference model HS3-NUVEL-1A. A previous global comparison of the SKS fast-polarization directions with flow models of the upper mantle showed relatively poor correlation on the continents, which was interpreted as evidence for a large contribution of ""frozen"" anisotropy in the lithosphere. For the South American plate, our data indicate that one of the reasons for the poor correlation may have been the relatively coarse model of lithospheric thicknesses. We suggest that improved models of upper-mantle flow that are based on more detailed lithospheric thicknesses in South America may help to explain most of the observed anisotropy patterns.

The stable archipelago in the region of the Pallas and Hansa dynamical families

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Monitoramento ambiental da costa do Rio Grande do Norte com base em sensoriamento remoto e geodésia de precisão

This paper presents models of parameters of Sea Surface Layer (SSL), such as chlorophyll-a, sea surface temperature (SST), Primary Productivity (PP) and Total Suspended Matter (TSM) for the region adjacent to the continental shelf of Rio Grande do Norte (RN), Brazil. Concentrations of these parameters measured in situ were compared in time quasi-synchronous with images AQUA-MODIS between the years 2003 to 2011. Determination coefficients between samples in situ and bands reflectance sensor AQUA-MODIS were representative. From that, concentrations of SSL parameters were acquired for the continental shelf of the RN (eastern and northern) analyzing the geographic distribution of variation of these parameters between the years 2009-2012. Geographical and seasonal variations mainly influenced by global climate phenomena such as El Niño and La Niña, were found through the analysis of AQUA-MODIS images by Principal Components Analysis (PCA). Images show qualitatively the variance and availability of TSM in the regions, as well as their relationship with coastal erosion hotspots, monitored along the coast of the RN. In one of the areas identified as being of limited availability of TSM, we developed a methodology for assessment and evaluation of Digital Elevation Models (DEM) of beach surfaces (emerged and submerged sections) from the integration of topographic and bathymetric data measured in situ and accurately georeferenced compatible to studies of geomorphology and coastal dynamics of short duration. The methodology consisted of surveys with GNSS positioning operated in cinematic relative mode involved in topographic and bathymetric executed in relation to the stations of the geodetic network of the study area, which provided geodetic link to the Brazilian Geodetic System (GBS), univocal , fixed, and relatively stable over time. In this study Ponta Negra Beach, Natal / RN, was identified as a region with low variance and availability of MPS in the region off, as characterized by intense human occupation and intense coastal erosion in recent decades, which presents potential of the proposed methodology for accuracy and productivity, and the progress achieved in relation to the classical methods of surveying beach profiles

Caracterização Morfodinâmica e Sedimentologia da Plataforma Continental Rasa na APA Estadual dos Recifes de Corais - RN

This study had to aimed to characterize the sediments of shallow continental shelf and realize the mapping of features visible for satellite images by using remote sensing techniques, digital image processing and analysis of bathymetry between Maxaranguape and Touros - RN. The study s area is located in the continental shallow shelf of Rio Grande do Norte, Brazil, and is part of the Environmental Protection Area (APA) of Coral Reefs. A total of 1186 sediment samples were collected using a dredge type van veen and positioning of the vessel was made out with the aid of a Garmin 520s. The samples were treated In the laboratory to analyze particle size of the sediment, concentration of calcium carbonate and biogenic composition. The digital images from the Landsat-5 TM were used to mapping of features. This stage was used the band 1 (0,45-1,52 μm) where the image were georeferenced, and then adjusting the histogram, giving a better view of feature bottom and contacts between different types of bottom. The results obtained from analysis of the sediment showed that the sediments of the continental shelf east of RN have a dominance of carbonate facies and a sand-gravelly bottom because the region is dominated by biogenic sediments, that are made mainly of calcareous algae. The bedform types identified and morphological features found were validated by bathymetric data and sediment samples examined. From the results obtained a division for the shelf under study is suggested, these regions being subdivided, in well characterized: (1) Turbid Zone, (2) Coral Patch Reefs Zone, (3) Mixed Sediments Carbonates Zone, ( 4) Algae Fouling Zone, (5) Alignment Rocky Zone, (6) Sand Waves Field (7) Deposit siliciclastic sands