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.     

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.     

A retrotransposon-mediated gene duplication underlies morphological variation of tomato fruit

Edible fruits, such as that of the tomato plant and other vegetable crops, are markedly diverse in shape and size. SUN, one of the major genes controlling the elongated fruit shape of tomato, was positionally cloned and found to encode a member of the IQ67 domain-containing family. We show that the locus arose as a result of an unusual 24.7-kilobase gene duplication event mediated by the long terminal repeat retrotransposon Rider. This event resulted in a new genomic context that increased SUN expression relative to that of the ancestral copy, culminating in an elongated fruit shape. Our discovery demonstrates that retrotransposons may be a major driving force in genome evolution and gene duplication, resulting in phenotypic change in plants.     

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.     

Survival of Penicillium bilaiae inoculated on canola seed treated with Vitavax RS and Extender

The fungicide Vitavax RS and the seed polymer Extender were sequentially applied to canola seed to determine their effect on the fungal inoculum, Penicillium bilaiae. In the laboratory, the fungicide was applied at the recommended rate of 23.4 ml kg^–1, and the polymer was applied by the manufacturer. Significant death of P. bilaiae occurred after 2.5 days on the Vitavax RS-treated seed and after 1.5 days with the seed treated with both Vitavax RS and Extender. In field treatments, the seed was sown at six sites in the fall just prior to freeze-up. Results from the laboratory experiment indicate that there should be no harmful effects of the Vitavax RS and Extender on the P. bilaiae at the time of seeding because the seeds were sown within 2 h of inoculation. The environmental extremes in 2001 and 2002 resulted in large variations in the data. P. bilaiae did not increase yield under these conditions.     

Assessment of Water Quality in Association with Land use in the Tillamook Bay Watershed, Oregon, USA

The water quality in tributaries to Tillamook Bay, Oregon, frequently exceeds standards for fecal coliform bacteria (FCB) and temperature. FCB inputs to the bay have forced periodic closure of the oyster shellfish industry. In addition, impaired water quality may be contributing to reduced salmonid populations in the bay and its tributaries through reduction in the quality of the habitat. Because of these concerns, the Tillamook Bay National Estuary Project (TBNEP) conducted several characterization studies and a long-term water quality monitoring program for the tributary rivers. This paper summarizes data collected to date within these efforts, including storm-based data on FCB and total suspended solids (TSS), and bimonthly data on nutrient concentrations in selected rivers. Monitoring data from 1996 to 2002 are summarized. Results for FCB and TSS are reported by storm, which are classified according to season and precipitation patterns in order to minimize intra-annual variability. There are not indications of large changes in water quality throughout the period of record, but it is too early in the program for trends analysis. Storms that exhibited the highest FCB concentrations tended to be those that occurred during fall and/or those that were preceded by relatively dry conditions and included high rainfall intensity. Implementation of storm-based monitoring and classification of storms according to season effectively reduces the large variability inherent in the FCB monitoring data, thereby facilitating future trends analysis. Continued storm-based monitoring of FCB and TSS, and also continued collection of rainfall and river discharge data, will provide the database that will be needed to determine to what extent on-the-ground remediation actions and best management practices (BMPs) within the Tillamook Basin are having their desired effects.     

An in vitro biomechanical comparison of the breaking strength and stiffness of polydioxanone (sizes 2, 7) and polyglactin 910 (sizes 3, 6) in the equine linea alba

OBJECTIVE: To determine failure strength, stiffness, and failure mode of 4 suture materials in equine linea alba. STUDY DESIGN: Randomized complete block design. SAMPLE POPULATION: Linea albas collected from 12 adult horses (mean weight, 475 kg; mean age, 10 years). METHODS: The ventral abdominal fascia, including the linea alba, was collected and bisected along the linea alba into right and left halves. Each half was divided into four, 5-cm sections extending from the umbilicus cranially, and randomly assigned to 1 of 4 suture materials: 2 polydioxanone (2 PD), 3 polyglactin 910 (3 PG), 6 polyglactin 910 (6 PG), and 7 polydioxanone (7 PD). A single cycle to failure test was performed on each specimen at a distraction rate of 100 mm/min. Differences in failure strength and stiffness for the materials were evaluated using a mixed linear model with significance set at P<.05. RESULTS: In 94 of 96 test sections, constructs failed by suture failure. There were significant differences in failure strength (P<.0001) and stiffness (P<.001) among the suture/tissue constructs. 7 PD had the highest breaking strength (316.8 N) followed by 6 PG (281.3 N), 3 PG (229.9 N), and 2 PD (193.0 N). Six PG had the largest stiffness (14 N/mm) followed by 3 PG (12.7 N/mm), 7 PD (10.1 N/mm), and 2 PD (7.2 N/mm). Suture breaking strength and stiffness were not affected by linea or fascia thickness, individual horse, half of the linea alba, or abdominal wall position. Eighty-five (90.4%) suture loops failed adjacent the knot. CONCLUSIONS: When tested in single cycle to failure, suture material was more likely to fail than the linea alba. Larger suture materials had higher breaking strengths than smaller suture materials and stiffness was also affected by suture material and size. CLINICAL RELEVANCE: Given its high initial breaking strength and its relatively longer in vivo strength retention, 7 PD seems an appropriate choice of suture material for closure of the equine linea alba when maximal short-term failure strength is desired.     

Effect of carboxymethylcellulose and hyaluronate solutions on jejunal healing in horses

OBJECTIVE: To compare a double-layer inverting anastomosis with a single-layer appositional anastomosis, coated with either 1% sodium carboxymethylcellulose (SCMC) or 0.4% sodium hyaluronate (HA) solutions, in the small intestine of horses with respect to anastomotic healing and adhesion formation. ANIMALS: 18 adult horses. PROCEDURE: Midline celiotomy and end-to-end jejunal anastomoses were performed. In control group horses (n = 6), a double-layer inverting anastomosis coated with sterile lactated Ringer's solution was performed. In treatment group horses, a single-layer appositional anastomosis was performed that was coated with 1% carboxymethylcellulose solution (SAA + SCMC group horses, 6) or 0.4% hyaluronate solution (SAA + HA group horses, 6). An additional 500 mL of the respective treatment solution was applied to the jejunal serosal surface, and 2 jejunal serosal abrasion sites were created. Horses were euthanatized 10 days after surgery. Anastomoses and abdominal adhesions were evaluated grossly. Anastomotic healing was evaluated on the basis of bursting wall tension. RESULTS: Bursting wall tension was significantly greater in SAA + SCMC group horses, compared with control group horses. All intestinal segments failed at a point distant to the anastomosis. Significantly fewer adhesions were found at the abrasion sites of SAA + HA group horses, compared with control group horses. No differences were found in adhesion formation at the anastomotic sites among groups. CONCLUSIONS AND CLINICAL RELEVANCE: Coating a single-layer appositional jejunal anastomosis with SCMC or HA solutions does not adversely affect anastomotic healing. Application of 0.4% HA solution to the serosal surface of the jejunum significantly decreases the incidence of experimentally induced intra-abdominal adhesion formation in horses.