Field and laboratory biology of the stem-feeding beetle Thamnurgus euphorbiae (Küster) (Col., Scolytidae) in Italy, a potential biological control candidate of leafy spurge in the USA and Canada

Thamnurgus euphorbiae overwinters as adults in dry stems of Euphorbia characias until the end of March before exiting through circular holes cut with their mandibles. Females and males appeared in the field at the same time, and the first pairs were observed on 28 March on new stems of E. characias having the open‐flower at the beginning of its development. Throughout the rest of April it was possible to find and collect the fairly abundant adults. Mating occurs on new stems of the host plant and females walk up towards the top of the stem and start to mine the centre of it in order to penetrate. Oviposition occurs in new stems and eggs are laid singly along the interior of the stem; females laid 35–85 eggs. Upon hatching, larvae started feeding on vascular bundles and the inner cortex. There are three larval instars, and pupation occurred in the stem of the host plant; this insect is univoltine. Thamnurgus euphorbiae has been accepted by the Technical Advisory Group for Biological Control Agents of Weeds for release as a biological control agent of leafy spurge in the US. Prior to this study the biology of T. euphorbiae was unknown.     

Herpesvirus salmonis: Characterization of a New Pathogen of Rainbow Trout

A new agent, provisionally designated Herpesvirus salmonis, was isolated from post-spawning rainbow trout (Salmo gairdneri) and studied primarily in the RTG-2 rainbow trout cell line. Infection of RTG-2 cells resulted in the formation of syncytia and Cowdry type A intranuclear inclusions. Replication occurred regularly at 5 and 10°C, but was inconsistent at 15°C, largely inhibited at 0°C, and completely inhibited at 20°C or higher. The virus was acid, heat, ether, and chloroform labile, but stable to freezing and thawing. It did not hemagglutinate. Viral DNA had a buoyant density of 1.709 g/cm³ and a guanine-cytosine value of 50%. Hexagonal nucleocapsids had a diameter of 90 nm and were first seen in nuclei at 36 h. Enveloped forms measured about 150 nm and occurred both cytoplasmically and extracellularly. At 10°C, a one-step growth culture required about 96 h; cell-associated virus peaked at about 10⁵ PFU/ml and exceeded released virus by a factor of about 10.     

Acepromazine-ketamine anesthesia in the rhesus monkey (Macaca mulatta).

Six adult male rhesus monkeys (Macaca mulatta) were anesthetized using a combination of acepromazine maleate and ketamine hydrochloride administered by intramuscular injection. This combination produced a smooth induction to anesthesia requiring less than 5 minutes. The average duration of anesthesia was slightly less than 1 hour and was safely prolonged with additional doses of ketamine. The depth of anesthesia was sufficient for minor surgical procedures and for precise radiological studies. No deleterious side effects were noted, and animals recovered completely in a short time.     

Effects of irradiance on relative growth rates, net assimilation rates, and leaf area partitioning in cotton and three associated weeds

Cotton (Gossypium hirsutum L. var. `Stoneville 213'), velvetleaf (Abutilon theophrasti Medic.), redroot pigweed (Amaranthus retroflexus L.), and hemp sesbania (Sesbania exaltata [Raf.] Cory) were grown in a controlled environment room at 31/25 C day/night temperature and three irradiances: 90, 320, and 750 μeinsteins meter⁻² second⁻¹. From total dry weights and leaf areas determined at intervals during the first exponential phase of growth, we used mathematical growth analysis techniques to calculate net assimilation rates (NAR), relative growth rates (R_w), relative leaf area expansion rates (R_a), leaf area partition coefficients (LAP), and leaf area ratios (LAR). In all four species, R_w, R_a, and NAR decreased with decreasing growth irradiance, while LAP and LAR increased. Within each species, R_w was positively correlated with NAR but negatively correlated with LAP and LAR. In comparisons among the four species within each growth irradiance, R_w was positively correlated with LAP. We discuss the relationship between LAP and LAR and show that LAP = (R_a/R_w) (LAR).     

Relationship Between Medium Salinity, Body Density, Buoyancy and Swimming in Artemia franciscana Larvae: Constraints on Water Column Use?

Body density measurements were carried out on larval Artemia franciscana that had been held for 24 h at either 33 or 100‰ after hatching. Body density was low (1.0308–1.0342 g ml⁻¹) by comparison with marine crustacean zooplankters or benthic freshwater crustaceans, and very stable, being only 0.3% higher in 100‰ than in 33‰. Vertical and horizontal swimming in 2nd instar larvae was studied at 8.5, 17, 34, 50, 100, 150 and 250‰ at 23 °C. At 8.5–50‰ downwards swimming was always significantly faster than upwards swimming, but in 100‰ downwards swimming was much slower than upwards swimming, indicating substantial positive buoyancy. At 150 and 250‰ downwards swimming was impossible. Horizontal swimming speed was unaffected by salinity over the range 8.5–100‰, but could not be studied at 150 and 250‰ as larvae could not leave the surface film. An asymptotic relationship between salinity and viscosity was found over the range 8.5–250‰. Calculations indicate a rise from 1.197 to 1.513 cp between 8.5 and 100‰, but this does not significantly impede horizontal locomotion. It appears that water column use by early stage Artemia larvae, that use a pair of 2nd antennae for rowing-type locomotion, is constrained by the increasing positive buoyancy associated with living at salinities above that corresponding to neutral buoyancy (approximately 48‰).     

Self-reported sugar-sweetened beverage intake among college students

Objective: To characterize sugar‐sweetened beverage intake of college students. Research Methods and Procedures: Undergraduates in an urban southern community campus were surveyed anonymously about sugared beverage consumption (soda, fruit drinks, energy drinks, sports drinks, sweet ice tea) in the past month. Results: Two hundred sixty‐five undergraduates responded (66% women, 46% minority, 100% of volunteers solicited). Most students (95%) reported sugared beverage intake in the past month, and 65% reported daily intake. Men were more likely than women to report daily intake (74% vs. 61%, p = 0.035). Soda was the most common sugar‐sweetened beverage. Black undergraduates reported higher sugared beverage intake than whites (p = 0.02), with 91% of blacks reporting sugar‐sweetened fruit drink intake in the past month and 50% reporting daily consumption. Mean estimated caloric intake from combined types of sugar‐sweetened beverages was significantly higher among black students than whites, 796 ± 941 vs. 397 ± 396 kcal/d (p = 0.0003); the primary source of sugar‐sweetened beverage calories among blacks was sugared fruit drinks (556 ± 918 kcal/d). Younger undergraduates reported significantly higher intake than older students (p = 0.025). Discussion: Self‐reported sugar‐sweetened beverage consumption among undergraduates is substantial and likely contributes considerable non‐nutritive calories, which may contribute to weight gain. Black undergraduates may be particularly vulnerable due to higher sugared beverage intake. Obesity prevention interventions targeting reductions in sugar‐sweetened beverages in this population merit consideration.     

Massively parallel characterization of restriction endonucleases

Restriction endonucleases are highly specific in recognizing the particular DNA sequence they act on. However, their activity is affected by sequence context, enzyme concentration and buffer composition. Changes in these factors may lead to either ineffective cleavage at the cognate restriction site or relaxed specificity allowing cleavage of degenerate ‘star’ sites. Additionally, uncharacterized restriction endonucleases and engineered variants present novel activities. Traditionally, restriction endonuclease activity is assayed on simple substrates such as plasmids and synthesized oligonucleotides. We present and use high-throughput Illumina sequencing-based strategies to assay the sequence specificity and flanking sequence preference of restriction endonucleases. The techniques use fragmented DNA from sequenced genomes to quantify restriction endonuclease cleavage on a complex genomic DNA substrate in a single reaction. By mapping millions of restriction site–flanking reads back to the Escherichia coli and Drosophila melanogaster genomes we were able to quantitatively characterize the cognate and star site activity of EcoRI and MfeI and demonstrate genome-wide decreases in star activity with engineered high-fidelity variants EcoRI-HF and MfeI-HF, as well as quantify the influence on MfeI cleavage conferred by flanking nucleotides. The methods presented are readily applicable to all type II restriction endonucleases that cleave both strands of double-stranded DNA.     

Bone morphogenetic protein signalling in airway epithelial cells during regeneration

Mechanisms of lung regeneration after injury remain poorly understood. Bone morphogenetic protein 4 (BMP4) is critical for lung morphogenesis and regulates differentiation of the airway epithelium during development, although its mechanism of action is unknown. The role of BMPs in adult lungs is unclear. We hypothesised that BMP signalling is involved in regeneration of damaged adult airways after injury. Our aims were to characterise the regeneration process in 1-nitronaphthalene (1-NN) injured airways, to determine if and when BMP signalling is activated during this process and investigate the effects of BMP4 on normal adult airway epithelial cells (AECs). Rats were injected with 50 mg/kg 1-NN and protein expression in AECs was examined by Western blotting of lung lysis lavage, and by immunofluorescence, at 6, 24, 48 and 96 h post injection. Expression of signalling molecules p-ERK-1, p-ERK-2 and p-Smad1/5/8 in AECs peaked at 6 h post injection, coincident with maximal inflammation and prior to airway denudation which occurred at 24 h. While airways were re-epithelialised by 48 h, AEC proliferation peaked later at 96 h post 1-NN injection. In vitro, BMP4 induced a mesenchymal-like morphology in normal AECs, downregulated E-cadherin expression and increased migration in a wound closure assay. Thus, following acute injury, increased BMP signalling in AECs coincides with inflammation and precedes airway denudation and re-epithelialisation. Our data indicate that, similar to its role in controlling tissue architecture during development, BMP signalling regulates regeneration of the airways following acute injury, involving downregulation of E-cadherin and induction of migration in AECs.