Stable, monomeric variants of lambda Cro obtained by insertion of a designed beta-hairpin sequence

lambda Cro is a dimeric DNA binding protein. Random mutagenesis and a selection for Cro activity have been used to identify the contacts between Cro subunits that are crucial for maintenance of a stably folded structure. To obtain equivalent contacts in a monomeric system, a Cro variant was designed and constructed in which the antiparallel beta-ribbon that forms the dimer interface was replaced by a beta-hairpin. The engineered monomer has a folded structure similar to wild type, is significantly more stable than wild type, and exhibits novel half-operator binding activity.     

Plasma pharmacokinetics and pharmacodynamics of alfaxalone in neonatal foals after an intravenous bolus of alfaxalone following premedication with butorphanol tartrate

ObjectiveTo determine the pharmacokinetics and pharmacodynamics of the neurosteroid anaesthetic, alfaxalone, in neonatal foals after a single intravenous (IV) injection of alfaxalone following premedication with butorphanol tartrate.Study designProspective experimental study.AnimalsFive clinically healthy Australian Stock Horse foals of mean ± SD age of 12 ± 3 days and weighing 67.3 ± 12.4 kg.MethodsFoals were premedicated with butorphanol (0.05 mg kg^?1 IV) and anaesthesia was induced 10 minutes later by IV injection with alfaxalone 3 mg kg^?1. Cardiorespiratory variables (pulse rate, respiratory rate, direct arterial blood pressure, arterial blood gases) and clinical signs of anaesthetic depth were evaluated throughout anaesthesia. Venous blood samples were collected at strategic time points and alfaxalone plasma concentrations were assayed using liquid chromatography-mass spectrometry (LC/MS) and analysed by noncompartmental pharmacokinetic analysis.ResultsThe harmonic, mean ± SD plasma elimination half life (t½) for alfaxalone was 22.8 ± 5.2 minutes. The observed mean plasma clearance (Cl_p) and volume of distribution (Vd) were 19.9 ± 5.9 mL minute kg^?1 and 0.6 ± 0.2 L kg^?1, respectively. Overall, the quality of the anaesthetic inductions and recoveries was good and most monitored physiological variables were clinically acceptable in all foals, although some foals became hypoxaemic for a short period following recumbency. The mean durations of anaesthesia from induction to first movement and from induction to standing were 18.7 ± 7 and 37.2 ± 4.7 minutes, respectively.ConclusionsThe anaesthetic protocol used provided a predictable and consistent plane of anaesthesia in the five foals studied, with minimal cardiovascular depression. In foals, as in the adult horse, alfaxalone has a short elimination half life.Clinical relevanceAlfaxalone appears to be an adequate anaesthetic induction agent in foals and the pharmacokinetics suggest that, with continuous infusion, it might be suitable to provide more prolonged anaesthesia. Oxygen supplementation is recommended.     

Soil carbon and tree litter dynamics in a red cedar–scotch pine shelterbelt

Carbon sequestration in the woody biomass of shelterbelts has been investigated but there have been no measurements of the C stocks in soil and tree litter under this agroforestry practice. The objective of this study was to quantify C stored in surface soil layers and tree litter within and adjacent to a 35-year-old shelterbelt in eastern Nebraska, USA. The 2-row shelterbelt was composed of eastern red cedar (Juniperus virginiana) and scotch pine (Pinus sylvestris). A sampling grid was established across a section of the shelterbelt on Tomek silt loam (fine, smectitic, mesic Pachic Argiudolls). Four soil cores were collected at each grid point, divided into 0–7.5 and 7.5–15 cm depth increments, and composited by depth. Soil samples were analyzed for total, organic, and inorganic C, total N, texture, pH, and nutrient content. Under the shelterbelt, all surface litter in a 0.5 × 0.5 m² area at each grid point was collected prior to soil sampling, dried, weighed, sorted, and analyzed for total C and N. Average soil organic carbon (SOC) in the 0–15 cm layer within the shelterbelt (3,994 g m⁻²) was significantly greater than in the cultivated fields (3,623 g m⁻²). The tree litter contained an additional ∼1,300 g C m⁻². Patterns of litter mass and soil pH and texture suggested increased organic inputs by tree litter and deposition of wind-blown sediment may be responsible for greater SOC beneath the shelterbelt. Further research is needed to identify the mechanism(s) responsible for the observed patterns of SOC within and adjacent to the shelterbelt and to quantify the C in biomass and deeper soil layers.

Stenotrophomonas maltophilia as a part of normal oral bacterial flora in captive snakes and its susceptibility to antibiotics

Only little is known about normal oral bacterial flora in captive snakes containing Stenotrophomonas maltophilia. This microbe has been reported as a causative agent of numerous infections in reptiles. Therefore, the goal of the study was to detect its presence in the mouths of a significant number of healthy captive snakes and determining its susceptibility to antibiotics at 30 and 37 degrees C. The isolates were obtained in 1999-2005 from mouth swabs of 115 snakes of 12 genera and 22 species-most often Elaphe guttata (24 individuals; 20.9%). Susceptibility to 24 antibiotics was tested by the microdilution method. The microbe was demonstrated in 34 (29.6%) individuals. Overall, 47 strains of S. maltophilia were acquired. Evaluation using PFGE profiles and antibiograms resulted in confirmation of one strain of S. maltophilia in 23 (20.0%) individuals, two strains in nine (7.8%) and three in two (1.8%) snakes. All tested antibiotics were more effective at 37 degrees C, with the partial exception of cotrimoxazole and cefoperazone/sulbactam. At a temperature of 37 degrees C, the lowest frequency of resistance to levofloxacin (no resistant strains), cotrimoxazole and ofloxacin (97.9% of susceptible strains) was recorded. At 30 degrees C, the most active agents were cotrimoxazole (97.9% of susceptible strains), levofloxacin (91.5%) and ofloxacin (85.1%). In conclusion, S. maltophilia is present in the mouths of about one third of healthy captive snakes, showing good susceptibility to cotrimoxazole, some fluoroquinolones and aminoglycosides. The antibiotics (particularly aminoglycosides) are more effective at 37 degrees C.     

Short Communication. Quantification of loess proportions in Pleistocene periglacial slope deposits and Holocene colluvium using grain‐size data by laser diffractometry

Loess proportions in slope deposits have rarely been quantified, due to the lack of quantification methods. In this study, we used laser granulometry to analyze grain‐size distributions in slope deposits of a Buntsandstein landscape in central Germany. The consistent loess contents obtained from two independent models corroborate the validity of the calculated loess proportions and the applicability of the new R package EMMAgeo. Yet, some limitations in transferability need to be considered.     

Multidimensional optimality of microbial metabolism

Although the network topology of metabolism is well known, understanding the principles that govern the distribution of fluxes through metabolism lags behind. Experimentally, these fluxes can be measured by (13)C-flux analysis, and there has been a long-standing interest in understanding this functional network operation from an evolutionary perspective. On the basis of (13)C-determined fluxes from nine bacteria and multi-objective optimization theory, we show that metabolism operates close to the Pareto-optimal surface of a three-dimensional space defined by competing objectives. Consistent with flux data from evolved Escherichia coli, we propose that flux states evolve under the trade-off between two principles: optimality under one given condition and minimal adjustment between conditions. These principles form the forces by which evolution shapes metabolic fluxes in microorganisms' environmental context.