Self-assembling protein microarrays

Protein microarrays provide a powerful tool for the study of protein function. However, they are not widely used, in part because of the challenges in producing proteins to spot on the arrays. We generated protein microarrays by printing complementary DNAs onto glass slides and then translating target proteins with mammalian reticulocyte lysate. Epitope tags fused to the proteins allowed them to be immobilized in situ. This obviated the need to purify proteins, avoided protein stability problems during storage, and captured sufficient protein for functional studies. We used the technology to map pairwise interactions among 29 human DNA replication initiation proteins, recapitulate the regulation of Cdt1 binding to select replication proteins, and map its geminin-binding domain.     

Temporal specificity in the cortical plasticity of visual space representation

The circuitry and function of mammalian visual cortex are shaped by patterns of visual stimuli, a plasticity likely mediated by synaptic modifications. In the adult cat, asynchronous visual stimuli in two adjacent retinal regions controlled the relative spike timing of two groups of cortical neurons with high precision. This asynchronous pairing induced rapid modifications of intracortical connections and shifts in receptive fields. These changes depended on the temporal order and interval between visual stimuli in a manner consistent with spike timing-dependent synaptic plasticity. Parallel to the cortical modifications found in the cat, such asynchronous visual stimuli also induced shifts in human spatial perception.     

Molecular layering of fluorinated ionic liquids at a charged sapphire (0001) surface

Room-temperature ionic liquids (RTILs) are promising candidates for a broad range of "green" applications, for which their interaction with solid surfaces plays a crucial role. In this high-energy x-ray reflectivity study, the temperature-dependent structures of three ionic liquids with the tris(pentafluoroethyl)trifluorophosphate anion in contact with a charged sapphire substrate were investigated with submolecular resolution. All three RTILs show strong interfacial layering, starting with a cation layer at the substrate and decaying exponentially into the bulk liquid. The observed decay length and layering period point to an interfacial ordering mechanism, akin to the charge inversion effect, which is suggested to originate from strong correlations between the unscreened ions. The observed layering is expected to be a generic feature of RTILs at charged interfaces.     

Deep Subseafloor Fungi as an Untapped Reservoir of Amphipathic Antimicrobial Compounds

The evolving global threat of antimicrobial resistance requires a deep renewal of the antibiotic arsenal including the isolation and characterization of new drugs. Underexplored marine ecosystems may represent an untapped reservoir of novel bioactive molecules. Deep-sea fungi isolated from a record-depth sediment core of almost 2000 m below the seafloor were investigated for antimicrobial activities. This antimicrobial screening, using 16 microbial targets, revealed 33% of filamentous fungi synthesizing bioactive compounds with activities against pathogenic bacteria and fungi. Interestingly, occurrence of antimicrobial producing isolates was well correlated with the complexity of the habitat (in term of microbial richness), as higher antimicrobial activities were obtained at specific layers of the sediment core. It clearly highlights complex deep-sea habitats as chemical battlefields where synthesis of numerous bioactive compounds appears critical for microbial competition. The six most promising deep subseafloor fungal isolates were selected for the production and extraction of bioactive compounds. Depending on the fungal isolates, antimicrobial compounds were only biosynthesized in semi-liquid or solid-state conditions as no antimicrobial activities were ever detected using liquid fermentation. An exception was made for one fungal isolate, and the extraction procedure designed to extract amphipathic compounds was successful and highlighted the amphiphilic profile of the bioactive metabolites.     

Ramifications of crop residue loading for soil microbial community composition,activity and nutrient supply

Variable results have been reported on the effects of crop residue loads on soil microbial properties. We investigated changes in soil bacterial composition, β-glucosidase enzyme activity and nutrient bioavailability in response to wheat residue loading. The treatments included three levels of above-ground wheat residues (removed, retained or supplemented), with or without fertilizer N. Bacteroidetes, Firmicutes and Verrucomicrobia (the first two are copiotrophs) were less abundant where residues were removed than where residues were retained or supplemented, but the reverse was true for Actinobacteria, Cyanobacteria, Chloroflexi and Nitrospirae (all oligotrophs, although some Actinobacteria can be copiotrophic). Actinobacteria were also less abundant where fertilizer N was applied, and the abundances of their genera (including Arthrobacter and Mycobacterium) increased where residues were removed, confirming that they were oligotrophic in this study. β-diversity showed similar differences in the bacterial community structures because of residue management, but α-diversity was not affected by residue management or N fertilizer. β-glucosidase enzyme activities increased as C inputs increased with residue manipulation and N fertilizer. The enzyme activities increased with increasing residue loading in the 0–15 cm soil depth, but decreased with soil depth. Soil K supply increased with increasing residue loading, but nitrate-N supply was highest with residue retention. These results demonstrate remarkable resilience of soil microbial functioning under a wide range of crop residue inputs, without adverse effects on enzyme activity attributable to inorganic N fertilizer. The increasing β-glucosidase activity with increasing residue loading probably explains why crop residue return does not always increase soil C stocks.     

Effects of graded doses of propofol for anesthesia induction on cardiovascular parameters and intraocular pressures in normal dogs

ObjectiveTo determine the effects of graded doses of propofol on cardiovascular parameters and intraocular pressures (IOP) in normal dogs.Study designProspective, randomized, modified Latin square, cross-over experimental study.AnimalsEleven adult random-source dogs weighing 20.2 ± 5.7 kg.MethodsThere were three treatment groups: propofol 8 mg kg^?1 intravenous (IV) until loss of jaw tone (Group P), propofol until loss of jaw tone +20% (Group P20), and propofol until loss of jaw tone +50% (Group P50). Atracurium 0.1 mg kg^?1 IV was administered in all treatments immediately after the propofol. All dogs received the three treatments in a randomized order, with at least a one week interval between treatments. Direct arterial blood pressure and IOP by applanation tonometry were obtained at baseline, after 5 minutes of pre-oxygenation (before induction), before, and after intubation. Blood gas samples were obtained at baseline, after pre-oxygenation, and before intubation.ResultsThere was no significant difference in IOP readings at any time point among groups. The IOP was significantly higher before intubation versus before induction in all three groups. There was a significantly smaller change in systolic, mean (MAP), and diastolic (DAP) arterial pressures in the P50 group compared with the P group after intubation. There was a significantly smaller change in MAP and DAP in the P50 group compared with the P20 group after intubation. The increase in CO₂ from before induction to before intubation was significantly greater in the P50 group than in the P or P20 groups.Conclusions and clinical relevanceGraded doses of propofol did not affect the increase in IOP observed with propofol induction in normal dogs. Higher doses of propofol are of no apparent additional benefit in animals who cannot tolerate an abrupt increase in IOP but may be of benefit in dogs who cannot tolerate an abrupt increase in blood pressure accompanying orotracheal intubation.