Single-cell mass cytometry of differential immune and drug responses across a human hematopoietic continuum

Flow cytometry is an essential tool for dissecting the functional complexity of hematopoiesis. We used single-cell "mass cytometry" to examine healthy human bone marrow, measuring 34 parameters simultaneously in single cells (binding of 31 antibodies, viability, DNA content, and relative cell size). The signaling behavior of cell subsets spanning a defined hematopoietic hierarchy was monitored with 18 simultaneous markers of functional signaling states perturbed by a set of ex vivo stimuli and inhibitors. The data set allowed for an algorithmically driven assembly of related cell types defined by surface antigen expression, providing a superimposable map of cell signaling responses in combination with drug inhibition. Visualized in this manner, the analysis revealed previously unappreciated instances of both precise signaling responses that were bounded within conventionally defined cell subsets and more continuous phosphorylation responses that crossed cell population boundaries in unexpected manners yet tracked closely with cellular phenotype. Collectively, such single-cell analyses provide system-wide views of immune signaling in healthy human hematopoiesis, against which drug action and disease can be compared for mechanistic studies and pharmacologic intervention.     

The BIOEXPLOIT Project

The EU Framework 6 Integrated Project BIOEXPLOIT concerns the exploitation of natural plant biodiversity for the pesticide-free production of food. It focuses on the pathogens Phytophthora infestans, Septoria tritici, Blumeria graminis, Puccinia spp. and Fusarium spp. and on the crops wheat, barley, tomato and potato. The project commenced in October 2005, comprises 45 laboratories in 12 countries, and is carried out by partners from research institutes, universities, private companies and small-medium enterprises. The project has four strategic objectives covered in eight sub-projects. These objectives relate to (i) understanding the molecular components involved in durable disease resistance, (ii) exploring and exploiting the natural biodiversity in disease resistance, (iii) accelerating the introduction of marker-assisted breeding and genetic engineering in the EU plant breeding industry, and (iv) coordinating and integrating resistance breeding research, providing training in new technologies, disseminating the results, and transferring knowledge and technologies to the industry.     

Rapid phase adjustment of melatonin and core body temperature rhythms following a 6-h advance of the light/dark cycle in the horse

Background  

Rapid displacement across multiple time zones results in a conflict between the new cycle of light and dark and the previously entrained program of the internal circadian clock, a phenomenon known as jet lag. In humans, jet lag is often characterized by malaise, appetite loss, fatigue, disturbed sleep and performance deficit, the consequences of which are of particular concern to athletes hoping to perform optimally at an international destination. As a species renowned for its capacity for athletic performance, the consequences of jet lag are also relevant for the horse. However, the duration and severity of jet lag related circadian disruption is presently unknown in this species. We investigated the rates of re-entrainment of serum melatonin and core body temperature (BT) rhythms following an abrupt 6-h phase advance of the LD cycle in the horse.     

Spatiotemporal Distribution of Two Euschistus spp. Stink Bugs (Hemiptera: Pentatomidae) in Southeastern Farmscapes

Stink bugs (Hemiptera: Pentatomidae) are ubiquitous, cryptic, phytophagous pests that are found in many crops. In agroecosystems, individuals disperse from adjacent noncrop hosts and tend to aggregate or cluster within fields. In this study, we characterized the distribution of Euschistus servus (Say) and Euschistus tristigmus (Say) (Hemiptera: Pentatomidae) over 2 yr at three southeastern United States farmscapes. Stink bugs were captured in pheromone-baited traps, and Spatial Analysis by Distance Indices (SADIE) used to identify the location of significant aggregations by habitat type and season. Euschistus servus adults were more likely to be captured in pecan orchards, cotton, other crops, or unmanaged habitats than in woodland habitats. Significant aggregations of E. servus were detected in a variety of habitats including pecan, corn, cotton, peanut, and tobacco, as well as fallow and hay fields, pastures, and hedgerows. Fewer adult E. tristigmus were captured than E. servus adults, and E. tristigmus adults were typically trapped and aggregated in woodland habitats. The resulting data provide an important understanding regarding the seasonal movement and relative abundance levels of stink bug populations, which are critical to the development of integrated pest management strategies.     

Food provisioning strategies,food insecurity,and stress in an economically vulnerable community: the Northern Cheyenne case

Living in poverty is associated with high levels of protracted stress associated with health problems. Economic and food insecurity are particularly poignant aspects of poverty and condition the work of securing basic daily needs of families. Recent studies suggest that levels of stress increase as family food needs rise. This paper presents new findings which clarify the relationship of food provisioning to stress levels, by examining actual food provisioning strategies and food insecurity among the Northern Cheyenne Indians of southeastern Montana. Results clearly show that stress varies by types of food acquisition strategies. Contrary to our expectations, more complex strategies, including relatively unpredictable and cumbersome food provisioning activities, are not linked to higher stress levels in our analysis. Controlling for food security levels, households using a combination of local programs and informal subsistence sources are the least stressed, despite the demands of managing a large number of food sources. Households primarily using Food Stamps are the most likely to experience high levels of stress. Interviews with Food Stamp recipients show that potential sources of stress include inadequate allocations of Food Stamps, difficulty achieving and maintaining eligibility, challenges to complying with paperwork and appointment requirements, as well as personal obstacles and community barriers to making food stamps last. Analyses indicate that contradictions between local cultural norms for food provisioning and the realities of food insecurity in this context promote strategies emphasizing greater independence from federal food programs.     

Regulation of blood and lymphatic vascular separation by signaling proteins SLP-76 and Syk

Lymphatic vessels develop from specialized endothelial cells in preexisting blood vessels, but the molecular signals that regulate this separation are unknown. Here we identify a failure to separate emerging lymphatic vessels from blood vessels in mice lacking the hematopoietic signaling protein SLP-76 or Syk. Blood-lymphatic connections lead to embryonic hemorrhage and arteriovenous shunting. Expression of slp-76 could not be detected in endothelial cells, and blood-filled lymphatics also arose in wild-type mice reconstituted with SLP-76-deficient bone marrow. These studies reveal a hematopoietic signaling pathway required for separation of the two major vascular networks in mammals.     

Structure of a gammadelta T cell receptor in complex with the nonclassical MHC T22

Gammadelta T cell receptors (TCRs), alphabeta TCRs, and antibodies are the three lineages of somatically recombined antigen receptors. The structural basis for ligand recognition is well defined for alphabeta TCR and antibodies but is lacking for gammadelta TCRs. We present the 3.4 A structure of the murine gammadelta TCR G8 bound to its major histocompatibility complex (MHC) class Ib ligand, T22. G8 predominantly uses germline-encoded residues of its delta chain complementarity-determining region 3 (CDR3) loop to bind T22 in an orientation substantially different from that seen in alphabeta TCR/peptide-MHC. That junctionally encoded G8 residues play an ancillary role in binding suggests a fusion of innate and adaptive recognition strategies.     

Evolution of block copolymer lithography to highly ordered square arrays

The manufacture of smaller, faster, more efficient microelectronic components is a major scientific and technological challenge, driven in part by a constant need for smaller lithographically defined features and patterns. Traditional self-assembling approaches based on block copolymer lithography spontaneously yield nanometer-sized hexagonal structures, but these features are not consistent with the industry-standard rectilinear coordinate system. We present a modular and hierarchical self-assembly strategy, combining supramolecular assembly of hydrogen-bonding units with controlled phase separation of diblock copolymers, for the generation of nanoscale square patterns. These square arrays will enable simplified addressability and circuit interconnection in integrated circuit manufacturing and nanotechnology.     

A critical role for eosinophils in allergic airways remodeling

Features of chronic asthma include airway hyperresponsiveness, inflammatory infiltrates, and structural changes in the airways, termed remodeling. The contribution of eosinophils, cells associated with asthma and allergy, remains to be established. We show that in mice with a total ablation of the eosinophil lineage, increases in airway hyperresponsiveness and mucus secretion were similar to those observed in wild-type mice, but eosinophil-deficient mice were significantly protected from peribronchiolar collagen deposition and increases in airway smooth muscle. These data suggest that eosinophils contribute substantially to airway remodeling but are not obligatory for allergen-induced lung dysfunction, and support an important role for eosinophil-targeted therapies in chronic asthma.     

Oxygen limitation and tissue metabolic potential of the African fish Barbus neumayeri : roles of native habitat and acclimatization

Background  

Oxygen availability in aquatic habitats is a major environmental factor influencing the ecology, behaviour, and physiology of fishes. This study evaluates the contribution of source population and hypoxic acclimatization of the African fish, Barbus neumayeri, in determining growth and tissue metabolic enzyme activities. Individuals were collected from two sites differing dramatically in concentration of dissolved oxygen (DO), Rwembaita Swamp (annual average DO 1.35 mgO₂ L^-1) and Inlet Stream West (annual average DO 5.58 mgO₂ L^-1) in Kibale National Park, Uganda, and reciprocally transplanted using a cage experiment in the field, allowing us to maintain individuals under natural conditions of oxygen, food availability, and flow. Fish were maintained under these conditions for four weeks and sampled for growth rate and the activities of phosphofructokinase (PFK), lactate dehydrogenase (LDH), citrate synthase (CS), and cytochrome c oxidase (CCO) in four tissues, liver, heart, brain, and skeletal muscle.