H+ transport is an integral function of the mitochondrial ADP/ATP carrier.

The mitochondrial ADP/ATP carrier (AAC) is a major transport protein of the inner mitochondrial membrane. It exchanges mitochondrial ATP for cytosolic ADP and controls cellular production of ATP. In addition, it has been proposed that AAC mediates mitochondrial uncoupling, but it has proven difficult to demonstrate this function or to elucidate its mechanisms. Here we record AAC currents directly from inner mitochondrial membranes from various mouse tissues and identify two distinct transport modes: ADP/ATP exchange and H+ transport. The AAC-mediated H+ current requires free fatty acids and resembles the H+ leak via the thermogenic uncoupling protein 1 found in brown fat. The ADP/ATP exchange via AAC negatively regulates the H+ leak, but does not completely inhibit it. This suggests that the H+ leak and mitochondrial uncoupling could be dynamically controlled by cellular ATP demand and the rate of ADP/ATP exchange. By mediating two distinct transport modes, ADP/ATP exchange and H+ leak, AAC connects coupled (ATP production) and uncoupled (thermogenesis) energy conversion in mitochondria

Terrestrial biosphere models need better representation of vegetation phenology: results from the North American Carbon Program Site Synthesis

Phenology, by controlling the seasonal activity of vegetation on the land surface, plays a fundamental role in regulating photosynthesis and other ecosystem processes, as well as competitive interactions and feedbacks to the climate system. We conducted an analysis to evaluate the representation of phenology, and the associated seasonality of ecosystem-scale CO2 exchange, in 14 models participating in the North American Carbon Program Site Synthesis. Model predictions were evaluated using long-term measurements (emphasizing the period 2000-2006) from 10 forested sites within the AmeriFlux and Fluxnet-Canada networks. In deciduous forests, almost all models consistently predicted that the growing season started earlier, and ended later, than was actually observed; biases of 2 weeks or more were typical. For these sites, most models were also unable to explain more than a small fraction of the observed interannual variability in phenological transition dates. Finally, for deciduous forests, misrepresentation of the seasonal cycle resulted in over-prediction of gross ecosystem photosynthesis by +160 ± 145 g C m-2 y-1 during the spring transition period, and +75 ± 130 g C m-2 y-1 during the autumn transition period (13% and 8% annual productivity, respectively) compensating for the tendency of most models to under-predict the magnitude of peak summertime photosynthetic rates. Models did a better job of predicting the seasonality of CO2 exchange for evergreen forests. These results highlight the need for improved understanding of the environmental controls on vegetation phenology, and incorporation of this knowledge into better phenological models. Existing models are unlikely to predict future responses of phenology to climate change accurately, and therefore will misrepresent the seasonality and interannual variability of key biosphere-atmosphere feedbacks and interactions in coupled global climate models.Engineering and Applied SciencesOrganismic and Evolutionary Biolog

Structural Basis of Cytotoxicity Mediated by the Type III Secretion Toxin ExoU from Pseudomonas aeruginosa

The type III secretion system (T3SS) is a complex macromolecular machinery employed by a number of Gram-negative pathogens to inject effectors directly into the cytoplasm of eukaryotic cells. ExoU from the opportunistic pathogen Pseudomonas aeruginosa is one of the most aggressive toxins injected by a T3SS, leading to rapid cell necrosis. Here we report the crystal structure of ExoU in complex with its chaperone, SpcU. ExoU folds into membrane-binding, bridging, and phospholipase domains. SpcU maintains the N-terminus of ExoU in an unfolded state, required for secretion. The phospholipase domain carries an embedded catalytic site whose position within ExoU does not permit direct interaction with the bilayer, which suggests that ExoU must undergo a conformational rearrangement in order to access lipids within the target membrane. The bridging domain connects catalytic domain and membrane-binding domains, the latter of which displays specificity to PI(4,5)P2. Both transfection experiments and infection of eukaryotic cells with ExoU-secreting bacteria show that ExoU ubiquitination results in its co-localization with endosomal markers. This could reflect an attempt of the infected cell to target ExoU for degradation in order to protect itself from its aggressive cytotoxic action

Fitness of Isogenic Colony Morphology Variants of Pseudomonas aeruginosa in Murine Airway Infection

Chronic lung infections with Pseudomonas aeruginosa are associated with the diversification of the persisting clone into niche specialists and morphotypes, a phenomenon called ‘dissociative behaviour’. To explore the potential of P. aeruginosa to change its morphotype by single step loss-of–function mutagenesis, a signature-tagged mini-Tn5 plasposon library of the cystic fibrosis airway isolate TBCF10839 was screened for colony morphology variants under nine different conditions in vitro. Transposon insertion into 1% of the genome changed colony morphology into eight discernable morphotypes. Half of the 55 targets encode features of primary or secondary metabolism whereby quinolone production was frequently affected. In the other half the transposon had inserted into genes of the functional categories transport, regulation or motility/chemotaxis. To mimic dissociative behaviour of isogenic strains in lungs, pools of 25 colony morphology variants were tested for competitive fitness in an acute murine airway infection model. Six of the 55 mutants either grew better or worse in vivo than in vitro, respectively. Metabolic proficiency of the colony morphology variant was a key determinant for survival in murine airways. The most common morphotype of self-destructive autolysis did unexpectedly not impair fitness. Transposon insertions into homologous genes of strain PAO1 did not reproduce the TBCF10839 mutant morphotypes for 16 of 19 examined loci pointing to an important role of the genetic background on colony morphology. Depending on the chosen P. aeruginosa strain, functional genome scans will explore other areas of the evolutionary landscape. Based on our discordant findings of mutant phenotypes in P. aeruginosa strains PAO1, PA14 and TBCF10839, we conclude that the current focus on few reference strains may miss modes of niche adaptation and dissociative behaviour that are relevant for the microevolution of complex traits in the wild

Fisheries as a source of marine debris on beaches in the United Kingdom

Marine debris from ships has persisted and remains a concern despite international agreements such as MARPOL. We report on an analysis of beach litter based on a data set established by the Marine Conservation Society (MSC) Beachwatch weekends. Debris collected around the UK was divided into three main types of debris: (1) plastic, (2) fishing, and (3) fishing related plastic and rubber. Correspondence analysis (CA) was used to examine patterns in the occurrence of debris types on a total of 1023 beaches and debris attributable to fishing was identified on clusters of beaches mainly located on the coasts of Scotland and along the English Channel. General Linear model (GLM) identified fishing as the highest explanatory factor when testing for relationships between litter and proximity to fishing ports and grounds. The results add to the growing body of evidence that the fishing industry is largely responsible for marine debris

\u3cem\u3eAlligator mississippiensis\u3c/em\u3e Sternal and Shoulder Girdle Mobility Increase Stride Length during High Walks

Crocodilians have played a significant role in evolutionary studies of archosaurs. Given that several major shifts in forelimb function occur within Archosauria, forelimb morphologies of living crocodilians are of particular importance in assessing locomotor evolutionary scenarios. A previous X-ray investigation of walking alligators revealed substantial movement of the shoulder girdle, but as the sternal cartilages do not show up in X-ray, the source of the mobility could not be conclusively determined. Scapulocoracoid movement was interpreted to indicate independent sliding of each coracoid at the sternocoracoid joint; however, rotations of the sternum could also produce similar displacement of the scapulocoracoids. Here, we present new data employing marker-based XROMM (X-ray reconstruction of moving morphology), wherein simultaneous biplanar X-ray video and surgically implanted radio-opaque markers permit precise measurement of the vertebral axis, sternum and coracoid in walking alligators. We found that movements of the sternum and sternocoracoid joint both contribute to shoulder girdle mobility and stride length, and that the sternocoracoid contribution was less than previously estimated. On average, the joint contributions to stride length (measured with reference to a point on the distal radius, thus excluding wrist motion) are as follows: thoracic vertebral rotation 6.2±3.7%, sternal rotation 11.1±2.5%, sternocoracoid joint 10.1±5.2%, glenohumeral joint 40.1±7.8% and elbow 31.1±4.2%. To our knowledge, this is the first evidence of sternal movement relative to the vertebral column (presumably via rib joints) contributing to stride length in tetrapods

Improving the process for evaluating the infrastructure in las Comunidades Especiales

This project, prepared for CSA Group of Architects and Engineers, presents the details for a standardized evaluation process for Special Communities. Through interviews with officials at the Special Communities Office and Department of Transportation and Public Works, CSA Group field architects and engineers, CSA Project Managers and GIS experts, we assessed the original evaluation process. The data from these interviews was collected and analyzed, resulting in the creation of a new standardized evaluation process which includes a new physical infrastructure assessment form. Both the evaluation process and form are to be used in the next two phases of the Special Communities Initiative