Retinoic Acid-Mediated Gene Expression in Transgenic Reporter Zebrafish

AbstractRetinoic acid-mediated gene activation is important for normal vertebrate development. The size and nature of retinoic acid make it difficult to identify the precise cellular location of this signaling molecule throughout an embryo. Additionally, retinoic acid (RA) signaling is regulated by a complex combination of receptors, coactivators, and antagonizing proteins. Thus, in order to integrate these signals and identify regions within a whole developing embryo where cells can respond transcriptionally to retinoic acid, we have used a reporter transgenic approach. We have generated several stable lines of transgenic zebrafish which use retinoic acid response elements to drive fluorescent protein expression. In these zebrafish lines, transgene expression is localized to regions of the neural tube, retina, notochord, somites, heart, pronephric ducts, branchial arches, and jaw muscles in embryos and larvae. Transgene expression can be induced in additional regions of the neural tube and retina as well as the immature notochord, hatching gland, enveloping cell layer, and fin by exposing embryos to retinoic acid. Treatment with retinoic acid synthase inhibitors, citral and diethylaminobenzaldehyde (DEAB), during neurulation, greatly reduces transgene expression. DEAB treatment of embryos at gastrulation phenocopies the embryonic effects of vitamin A deprivation or targeted disruption of the RA synthase retinaldehyde dehydrogenase-2 in other vertebrates. Together these data suggest that the reporter expression we see in zebrafish is dependent upon conserved vertebrate pathways of RA synthesis

Structural Changes in Isometrically Contracting Insect Flight Muscle Trapped following a Mechanical Perturbation

The application of rapidly applied length steps to actively contracting muscle is a classic method for synchronizing the response of myosin cross-bridges so that the average response of the ensemble can be measured. Alternatively, electron tomography (ET) is a technique that can report the structure of the individual members of the ensemble. We probed the structure of active myosin motors (cross-bridges) by applying 0.5% changes in length (either a stretch or a release) within 2 ms to isometrically contracting insect flight muscle (IFM) fibers followed after 5–6 ms by rapid freezing against a liquid helium cooled copper mirror. ET of freeze-substituted fibers, embedded and thin-sectioned, provides 3-D cross-bridge images, sorted by multivariate data analysis into ∼40 classes, distinct in average structure, population size and lattice distribution. Individual actin subunits are resolved facilitating quasi-atomic modeling of each class average to determine its binding strength (weak or strong) to actin. ∼98% of strong-binding acto-myosin attachments present after a length perturbation are confined to “target zones” of only two actin subunits located exactly midway between successive troponin complexes along each long-pitch helical repeat of actin. Significant changes in the types, distribution and structure of actin-myosin attachments occurred in a manner consistent with the mechanical transients. Most dramatic is near disappearance, after either length perturbation, of a class of weak-binding cross-bridges, attached within the target zone, that are highly likely to be precursors of strong-binding cross-bridges. These weak-binding cross-bridges were originally observed in isometrically contracting IFM. Their disappearance following a quick stretch or release can be explained by a recent kinetic model for muscle contraction, as behaviour consistent with their identification as precursors of strong-binding cross-bridges. The results provide a detailed model for contraction in IFM that may be applicable to contraction in other types of muscle

Antibiotic Use in Hispanic Households, New York City

Trained interviewers visited 631 inner city households to determine community prevalence and predictors of antibiotic use. Infectious disease symptoms were reported in 911 (33.2%) of 2,743 household members in the previous 30 days: medical attention was sought by 441 (48.4%) of 911 persons, and 354 (38.9%) of 911 took antibiotics for symptoms. Reported symptoms were respiratory (68.9%), gastrointestinal (15.3%), fever (12.8%), and skin infection (2.8%). Medical attention was sought significantly more often among those with chronic illness, those born in the United States, and those with fever, runny nose, or skin infections (all p<0.05). Antibiotics were taken significantly more often among those with poor health, those who spent more time at home, and those with fever and respiratory symptoms. Interventions to promote judicious use of antibiotics must include clinicians and the public, and for the Hispanic population such interventions must also be culturally relevant and provided in Spanish

The neotropical reforestation hotspots : a biophysical and socioeconomic typology of contemporary forest expansion

Tropical reforestation is a significant component of global environmental change that is far less understood than tropical deforestation, despite having apparently increased widely in scale during recent decades. The regional contexts defining such reforestation have not been well described. They are likely to differ significantly from the geographical profiles outlined by site-specific observations that predominate in the literature. In response, this article determines the distribution, extent, and defining contexts of apparently spontaneous reforestation. It delineates regional ‘hotspots’ of significant net reforestation across Latin America and the Caribbean and defines a typology of these hotspots with reference to the biophysical and socioeconomic characteristics that unite and distinguish amongst them. Fifteen regional hotspots were identified on the basis of spatial criteria pertaining to the area, distribution, and rate of reforestation 2001–2014, observed using a custom continental MODIS satellite land-cover classification. Collectively, these hotspots cover 11% of Latin America and the Caribbean and they include 167,667.7 km2 of new forests. Comparisons with other remotely sensed estimates of reforestation indicate that these hotspots contain a significant amount of tropical reforestation, continentally and pantropically. The extent of reforestation as a proportion of its hotspot was relatively invariable (3–14%) given large disparities in hotspot areas and contexts. An ordination analysis defined a typology of five clusters, distinguished largely by their topographical roughness and related aspects of agro-ecological marginality, climate, population trends, and degree of urbanization: ‘Urban lowlands’, ‘Mountainous populated areas’, ‘Rural highlands’, ‘Rural humid lands’ and ‘Rural dry lands’. The typology highlights that a range of distinct, even oppositional regional biophysical, demographic, and agricultural contexts have equally given rise to significant, regional net reforestation, urging a concomitant diversification of forest transition science

Tropomyosin controls sarcomere-like contractions for rigidity sensing and suppressing growth on soft matrices

Cells test the rigidity of the extracellular matrix by applying forces to it through integrin adhesions. Recent measurements show that these forces are applied via local micrometre-scale contractions, but how contraction force is regulated by rigidity is unknown. Here we performed high temporal- and spatial-resolution tracking of contractile forces by plating cells on sub-micron elastomeric pillars. We found that actomyosin-based sarcomere-like contractile units (CUs) simultaneously moved opposing pillars in net steps of ~2.5 nm, independent of rigidity. What correlated with rigidity was the number of steps taken to reach a force level that activated recruitment of α-actinin to the CUs. When we removed actomyosin restriction by depleting tropomyosin 2.1, we observed larger steps and higher forces that resulted in aberrant rigidity sensing and growth of non-transformed cells on soft matrices. Thus, we conclude that tropomyosin 2.1 acts as a suppressor of growth on soft matrices by supporting proper rigidity sensing

Optimisation of Perioperative Cardiovascular Management to Improve Surgical Outcome II (OPTIMISE II) trial: study protocol for a multicentre international trial of cardiac output-guided fluid therapy with low-dose inotrope infusion compared with usual care in patients undergoing major elective gastrointestinal surgery.

INTRODUCTION: Postoperative morbidity and mortality in older patients with comorbidities undergoing gastrointestinal surgery are a major burden on healthcare systems. Infections after surgery are common in such patients, prolonging hospitalisation and reducing postoperative short-term and long-term survival. Optimal management of perioperative intravenous fluids and inotropic drugs may reduce infection rates and improve outcomes from surgery. Previous small trials of cardiac-output-guided haemodynamic therapy algorithms suggested a modest reduction in postoperative morbidity. A large definitive trial is needed to confirm or refute this and inform widespread clinical practice. METHODS: The Optimisation of Perioperative Cardiovascular Management to Improve Surgical Outcome II (OPTIMISE II) trial is a multicentre, international, parallel group, open, randomised controlled trial. 2502 high-risk patients undergoing major elective gastrointestinal surgery will be randomly allocated in a 1:1 ratio using minimisation to minimally invasive cardiac output monitoring to guide protocolised administration of intravenous fluid combined with low-dose inotrope infusion, or usual care. The trial intervention will be carried out during and for 4 hours after surgery. The primary outcome is postoperative infection of Clavien-Dindo grade II or higher within 30 days of randomisation. Participants and those delivering the intervention will not be blinded to treatment allocation; however, outcome assessors will be blinded when feasible. Participant recruitment started in January 2017 and is scheduled to last 3 years, within 50 hospitals worldwide. ETHICS/DISSEMINATION: The OPTIMISE II trial has been approved by the UK National Research Ethics Service and has been approved by responsible ethics committees in all participating countries. The findings will be disseminated through publication in a widely accessible peer-reviewed scientific journal. TRIAL REGISTRATION NUMBER: ISRCTN39653756.The OPTIMISE II trial is supported by Edwards Lifesciences (Irvine, CA) and the UK National Institute for Health Research through RMP’s NIHR Professorship

African development corridors intersect key protected areas

[Extract] Unprecedented growth of mining and agriculture in Africa is encroaching on remaining habitats. Mining in Africa frequently occurs in proximity to protected areas (PAs), more than in other world regions (Durán, Rauch & Gaston, 2013), and at least 23 African PAs have been degazetted or downgraded as a result (Edwards et al., 2014). Even natural World Heritage Sites, the global pinnacle of conservation, suffer mining and oil/gas exploration and exploitation across 31 sites and 18 African countries (WWF, 2015), again more than other world regions (WWF, 2016). The anticipated expansion of transport 'development corridors' related to infrastructure and resource development could impact the ecological integrity of many other PAs as roads and rails link producers with refineries and ports over vast distances (Weng et al.,2013)