A conditional Mdm2 allele shows the importance of Mdm2 in heart development

Over 50% of sporadic tumors in humans have a p53 mutation highlighting its importance as a tumor suppressor. Considering additional mutations in other genes involved in p53 pathways, every tumor probably has mutant p53 or impaired p53-mediated functions. In response to a variety of cellular and genotoxic stresses, p53, mainly through its transcriptional activity, induces pathways involved in apoptosis and growth arrest. In these circumstances and under normal situations, p53 must be tightly regulated. Mdm2 is an important regulator of p53. Mdm2 inhibits p53 function by binding and blocking its transactivation domain. In addition, Mdm2 helps target p53 for degradation through its E3 ligase activity. Mdm2 null mice are embryonic lethal due to apoptosis in the blastocysts. However, a p53 null background rescues this lethality demonstrating the importance of the p53-Mdm2 interaction, particularly during development. The lethality of the Mdm2 null mouse prior to implantation limits the ability to investigate the role of Mdm2 in regulating p53 in a temporal and tissue specific manner. Does p53 need to be regulated in all tissues throughout the life of a mouse? Does Mdm2 always have to regulate it? To address these questions, we created a conditional Mdm2 allele. The conditional allele, Mdm2FM, in the presence of Cre recombinase results in the deletion of exons 5 and 6 of Mdm2 (most of the p53 binding domain) and represents a null allele. ^ The Mdm2FM allele was crossed with a heart muscle specific Cre expressing mouse (α-myosin heavy chain promoter driven Cre) to ask whether Mdm2 acts as a negative regulator of p53 in the heart. The heart is the most prominent organ early in embryogenesis and is shaped by cell death and proliferation. p53 does not appear to be active in the heart in response to some types of stress, so it remained to be determined if it has to be regulated in normal heart development. Loss of Mdm2 in the heart results in heart defects as early as E9.5. Loss of Mdm2 results in stabilized p53 and apoptosis. This apoptosis leads to a thinning of the myocardial wall particularly in the ventricles and abnormal ventricular structure. Eventually the abnormal heart fails resulting in lethality by E13.5. The embryonic lethality is rescued in a p53 null background. Thus, Mdm2 is important in regulating p53 in the development of the heart. ^

Molecular differentiation by PCR-RFLP technique shows that Antarctic fishes of the genus Notothenia, claimed to be two species, are phenotypic varieties with the same genetic pattern (Table 1)

The taxonomy of Antarctic fishes has been predominantly based on morphological characteristics rather than on genetic criteria. A typical example is the Notothenia group, which includes N. coriiceps Richardson, 1844, N. neglecta Nybelin, 1951 and N. rossii Richardson, 1844. The Polymerase Chain Reaction and Restriction Fragment Length Polymorphism (PCR-RFLP) technique was used to determine whether N. coriiceps Richardson, 1844 and N. neglecta Nybelin, 1951 are different or whether they are the same species with morphological, physiological and behavioural variability. N. rossii was used as control. Mitochondrial DNA (mtDNA) was isolated from muscle specimens of N. coriiceps Richardson, 1844, N. neglecta Nybelin, 1951 and N. rossii, which were collected in Admiralty Bay, King George Island. The DNA was used to amplify a fragment (690 base pairs) of the mitochondrial gene coding region of NADH dehydrogenase subunit 2. Further, the amplicon was digested with the following restriction enzymes: DdeI, HindIII and RsaI. The results showed a variation of the digestion pattern of the fragment amplified between N. rossii, and N. coriiceps Richardson, 1844 or N. neglecta Nybelin, 1951. However, no differences were found between N. coriiceps Richardson, 1844 and N. neglecta Nybelin, 1951, on the grounds of the same genetic pattern shown by the two fish.

Wal-Mart, caballo de Troya de la globalización

Creada en 1962 por Sam Walton, una pequeña empresa de distribución norteamericana se convertirá rápidamente en la primera empresa mundial. Desde Bentonville, Arkansas, hasta el interior de China, Wal-Mart muestra un camino bastante clásico que va desde la sede principal de la empresa hacia una proliferación de sus tiendas en una quincena de países. Con una facturación de 405 mil millones de dólares en 2008, la empresa de distribución número uno en el mundo fue sobrepasada en abril de 2009 por la empresa petrolera Exxon Mobil en la clasificación de las quinientas empresas mundiales más importantes. Pero a diferencia de otras grandes transnacionales como Exxon Mobil o Microsoft, Wal-Mart, la primera empresa mundial de distribución, tiene que adaptarse al medio ambiente local con el fin de atraer a un máximo de consumidores. El artículo propone una aproximación a escala múltiple de las estrategias de desarrollo de esta firma internacional. Así, la aplicación de un análisis espacial es apropiada para esclarecer los vínculos existentes entre los territorios y las estrategias de los actores a diferentes escalas geográficas.

Wal-Mart, caballo de Troya de la globalización

Creada en 1962 por Sam Walton, una pequeña empresa de distribución norteamericana se convertirá rápidamente en la primera empresa mundial. Desde Bentonville, Arkansas, hasta el interior de China, Wal-Mart muestra un camino bastante clásico que va desde la sede principal de la empresa hacia una proliferación de sus tiendas en una quincena de países. Con una facturación de 405 mil millones de dólares en 2008, la empresa de distribución número uno en el mundo fue sobrepasada en abril de 2009 por la empresa petrolera Exxon Mobil en la clasificación de las quinientas empresas mundiales más importantes. Pero a diferencia de otras grandes transnacionales como Exxon Mobil o Microsoft, Wal-Mart, la primera empresa mundial de distribución, tiene que adaptarse al medio ambiente local con el fin de atraer a un máximo de consumidores. El artículo propone una aproximación a escala múltiple de las estrategias de desarrollo de esta firma internacional. Así, la aplicación de un análisis espacial es apropiada para esclarecer los vínculos existentes entre los territorios y las estrategias de los actores a diferentes escalas geográficas.

Wal-Mart, caballo de Troya de la globalización

Creada en 1962 por Sam Walton, una pequeña empresa de distribución norteamericana se convertirá rápidamente en la primera empresa mundial. Desde Bentonville, Arkansas, hasta el interior de China, Wal-Mart muestra un camino bastante clásico que va desde la sede principal de la empresa hacia una proliferación de sus tiendas en una quincena de países. Con una facturación de 405 mil millones de dólares en 2008, la empresa de distribución número uno en el mundo fue sobrepasada en abril de 2009 por la empresa petrolera Exxon Mobil en la clasificación de las quinientas empresas mundiales más importantes. Pero a diferencia de otras grandes transnacionales como Exxon Mobil o Microsoft, Wal-Mart, la primera empresa mundial de distribución, tiene que adaptarse al medio ambiente local con el fin de atraer a un máximo de consumidores. El artículo propone una aproximación a escala múltiple de las estrategias de desarrollo de esta firma internacional. Así, la aplicación de un análisis espacial es apropiada para esclarecer los vínculos existentes entre los territorios y las estrategias de los actores a diferentes escalas geográficas.

Size matters: plasticity in metabolic scaling shows body-size may modulate responses to climate change

Variability in metabolic scaling in animals, the relationship between metabolic rate ( R) and body mass ( M), has been a source of debate and controversy for decades. R is proportional to Mb, the precise value of b much debated, but historically considered equal in all organisms. Recent metabolic theory, however, predicts b to vary among species with ecology and metabolic level, and may also vary within species under different abiotic conditions. Under climate change, most species will experience increased temperatures, and marine organisms will experience the additional stressor of decreased seawater pH ('ocean acidification'). Responses to these environmental changes are modulated by myriad species-specific factors. Body-size is a fundamental biological parameter, but its modulating role is relatively unexplored. Here, we show that changes to metabolic scaling reveal asymmetric responses to stressors across body-size ranges; b is systematically decreased under increasing temperature in three grazing molluscs, indicating smaller individuals were more responsive to warming. Larger individuals were, however, more responsive to reduced seawater pH in low temperatures. These alterations to the allometry of metabolism highlight abiotic control of metabolic scaling, and indicate that responses to climate warming and ocean acidification may be modulated by body-size.

Sea urchin response to rising pCO2 shows ocean acidification may fundamentally alter the chemistry of marine skeletons

Ocean acidification caused by an increase in pCO2 is expected to drastically affect marine ecosystem composition, yet there is much uncertainty about the mechanisms through which ecosystems may be affected. Here we studied sea urchins that are common and important grazers in the Mediterranean (Paracentrotus lividus and Arbacia lixula). Our study included a natural CO2 seep plus reference sites in the Aegean Sea off Greece. The distribution of A. lixula was unaffected by the low pH environment, whereas densities of P. lividus were much reduced. There was skeletal degradation in both species living in acidified waters compared to reference sites and remarkable increases in skeletal manganese levels (P. lividus had a 541% increase, A. lixula a 243% increase), presumably due to changes in mineral crystalline structure. Levels of strontium and zinc were also altered. It is not yet known whether such dramatic changes in skeletal chemistry will affect coastal systems but our study reveals a mechanism that may alter inter-species interactions.

Uso de DES y RANS no estacionario para la simulación del flujo alrededor de una carena Wigley - Use of Unsteady RANS and DES to simulate flows around a Wigley Hull)

Computer Fluid Dynamics tools have already become a valuable instrument for Naval Architects during the ship design process, thanks to their accuracy and the available computer power. Unfortunately, the development of RANSE codes, generally used when viscous effects play a major role in the flow, has not reached a mature stage, being the accuracy of the turbulence models and the free surface representation the most important sources of uncertainty. Another level of uncertainty is added when the simulations are carried out for unsteady flows, as those generally studied in seakeeping and maneuvering analysis and URANS equations solvers are used. Present work shows the applicability and the benefits derived from the use of new approaches for the turbulence modeling (Detached Eddy Simulation) and the free surface representation (Level Set) on the URANS equations solver CFDSHIP-Iowa. Compared to URANS, DES is expected to predict much broader frequency contents and behave better in flows where boundary layer separation plays a major role. Level Set methods are able to capture very complex free surface geometries, including breaking and overturning waves. The performance of these improvements is tested in set of fairly complex flows, generated by a Wigley hull at pure drift motion, with drift angle ranging from 10 to 60 degrees and at several Froude numbers to study the impact of its variation. Quantitative verification and validation are performed with the obtained results to guarantee their accuracy. The results show the capability of the CFDSHIP-Iowa code to carry out time-accurate simulations of complex flows of extreme unsteady ship maneuvers. The Level Set method is able to capture very complex geometries of the free surface and the use of DES in unsteady simulations highly improves the results obtained. Vortical structures and instabilities as a function of the drift angle and Fr are qualitatively identified. Overall analysis of the flow pattern shows a strong correlation between the vortical structures and free surface wave pattern. Karman-like vortex shedding is identified and the scaled St agrees well with the universal St value. Tip vortices are identified and the associated helical instabilities are analyzed. St using the hull length decreases with the increase of the distance along the vortex core (x), which is similar to results from other simulations. However, St scaled using distance along the vortex cores shows strong oscillations compared to almost constants for those previous simulations. The difference may be caused by the effect of the free-surface, grid resolution, and interaction between the tip vortex and other vortical structures, which needs further investigations. This study is exploratory in the sense that finer grids are desirable and experimental data is lacking for large α, especially for the local flow. More recently, high performance computational capability of CFDSHIP-Iowa V4 has been improved such that large scale computations are possible. DES for DTMB 5415 with bilge keels at α = 20º were conducted using three grids with 10M, 48M and 250M points. DES analysis for flows around KVLCC2 at α = 30º is analyzed using a 13M grid and compared with the results of DES on the 1.6M grid by. Both studies are consistent with what was concluded on grid resolution herein since dominant frequencies for shear-layer, Karman-like, horse-shoe and helical instabilities only show marginal variation on grid refinement. The penalties of using coarse grids are smaller frequency amplitude and less resolved TKE. Therefore finer grids should be used to improve V&V for resolving most of the active turbulent scales for all different Fr and α, which hopefully can be compared with additional EFD data for large α when it becomes available.

Effects of sheep and horse manure and pine bark amendments on metal distribution and chemical properties of contaminated mine soils.

Mine soils usually contain large levels of heavy metals and poor fertility conditions which limit their reclamation and the application of phyto-remediation technologies. Two organic waste materials (pine bark compost and sheep and horse manure compost), with different pHs and varying degrees of humification and nutrient contents, were applied as amendments to assess their effects on copper (Cu) and zinc (Zn) bioavailability and on fertility conditions of mine soils. Soil samples collected from two abandoned mining areas near Madrid (Spain) were mixed with 0, 30 and 60 t ha?1 of the organic amendments. The concentrations of metals among the different mineral and organic fractions of soil were determined by several extraction procedures to study the metal distribution in the solid phase of the soil affected by the organic amendments. The results showed that the manure amendment increased the soil pH and the cation exchange capacity and enhanced the nutrient levels of these soils. The pine bark amendment decreased the soil pH and did not significantly change the nutrient status of soil. Soil pH, organic matter content and its degree of humification, which were altered by the amendments, were the main factors affecting Cu fractionation. Zn fractionation was mainly affected by soil pH. The addition of manure not only improved soil fertility, but also decreased metal bioavailability resulting in a reduction of metal toxicity. Conversely, pine bark amendment increased metal ioavailability. The use of sheep and horse manure could be a cost-effective practice for the restoration of contaminated mine soils.

The 2.0-Å resolution crystal structure of a trimeric antibody fragment with noncognate VH–VL domain pairs shows a rearrangement of VH CDR3

The 2.0-Å resolution x-ray crystal structure of a novel trimeric antibody fragment, a “triabody,” has been determined. The trimer is made up of polypeptides constructed in a manner identical to that previously described for some “diabodies”: a VL domain directly fused to the C terminus of a VH domain—i.e., without any linker sequence. The trimer has three Fv heads with the polypeptides arranged in a cyclic, head-to-tail fashion. For the particular structure reported here, the polypeptide was constructed with a VH domain from one antibody fused to the VL domain from an unrelated antibody giving rise to “combinatorial” Fvs upon formation of the trimer. The structure shows that the exchange of the VL domain from antibody B1-8, a Vλ domain, with the VL domain from antibody NQ11, a Vκ domain, leads to a dramatic conformational change in the VH CDR3 loop of antibody B1-8. The magnitude of this change is similar to the largest of the conformational changes observed in antibody fragments in response to antigen binding. Combinatorial pairing of VH and VL domains constitutes a major component of antibody diversity. Conformationally flexible antigen-binding sites capable of adapting to the specific CDR3 loop context created upon VH–VL pairing may be employed by the immune system to maximize the structural diversity of the immune response.