Divalent copper complexes as influenza A M2 inhibitors.

New M2 blockers effective against the ubiquitous amantadine-resistant S31N M2 mutation in influenza A are needed. Six copper complexes, 2, 4, 6, 8, 9, and 10, were synthesized and found to block both wild type and S31N M2. Free Cu2+ also blocks M2 S31N but not S31N/H37A. The copper complexes do not block M2 H37A (either S31 or S31N). The complexes were effective against three influenza A strains in cell-culture assays, but less toxic to cells than CuCl2. For example 4, Cu(cyclooctylamineiminodiacetate), which was stable at pH > 4 in the buffers used, had an EC50 against A/Calif/07/2009 H1N1 of 0.7 ± 0.1 µM with a CC50 of 147 µM (therapeutic index, averaged over three strains, 67.8). In contrast, CuCl2 had an EC50 of 3.8 ± 0.9 µM and CC50 of 19 µM. Because M2 H37 is highly conserved, these complexes show promise for further testing as drugs against all strains of influenza A.

Heavy metal tolerance genes alter cellular thermodynamics in Pseudomonas putida and river Pseudomonas spp. and influence amebal predation.

Predation rates were measured for two Acanthamoeba castellanii strains feeding on metal-tolerant and metal-sensitive strains of Pseudomonas putida and compared with cellular thermodynamic data. Predation rates by A. castellanii strain ATCC 30010 correlated with cell volume of the prey. To explore whether this observation could be environmentally relevant, pseudomonad species were isolated from a pristine and a metal-contaminated river and were paired based on phylogenetic and physiological relatedness. Then, cellular thermodynamics and predation rates were measured on the most similar pseudomonad pair. Under cadmium stress, the strain from contaminated river sediments, Pseudomonas sp. CF150, exited metabolic dormancy faster than its pair from pristine sediments, Pseudomonas sp. N9, but consumed available resources less efficiently (more energy was lost as heat). Predation rates by both strains of ameba were greater on Pseudomonas sp. CF150 than on Pseudomonas sp. N9 at the highest cadmium concentration.

Pedobacter nyackensis sp. nov., Pedobacter alluvionis sp. nov. and Pedobacter borealis sp. nov., isolated from Montana flood-plain sediment and forest soil.

Three Gram-negative, rod-shaped, non-spore-forming eubacterial strains were isolated in western Montana, USA, and subjected to taxonomic studies. Strains NWG-II14(T) and NWER-II11(T) were isolated from hyporheic sediments of a large alluvial flood plain, whereas strain G-1(T) was isolated from a conifer forest soil. On the basis of 16S rRNA gene sequence similarity, strains NWG-II14(T), NWER-II11(T) and G-1(T) were shown to belong to the family Sphingobacteriaceae and are most closely related to various species of the genus Pedobacter. The results of molecular, physiological and biochemical tests allowed genotypic and phenotypic differentiation of these three strains from 23 Pedobacter species with validly published names. The three isolates therefore represent novel species, for which the names Pedobacter nyackensis sp. nov. (type strain NWG-II14(T) =DSM 19625(T) =LMG 24260(T)), Pedobacter alluvionis sp. nov. (type strain NWER-II11(T) =DSM 19624(T) =LMG 24258(T)) and Pedobacter borealis sp. nov. (type strain G-1(T) =DSM 19626(T) =LMG 24259(T)) are proposed.

Habitat heterogeneity and associated microbial community structure in a small-scale floodplain hyporheic flow path.

The Nyack floodplain is located on the Middle Fork of the Flathead River, an unregulated, pristine, fifth-order stream in Montana, USA, bordering Glacier National Park. The hyporheic zone is a nutritionally heterogeneous floodplain component harboring a diverse array of microbial assemblages essential in fluvial biogeochemical cycling, riverine ecosystem productivity, and trophic interactions. Despite these functions, microbial community structure in pristine hyporheic systems is not well characterized. The current study was designed to assess whether physical habitat heterogeneity within the hyporheic zone of the Nyack floodplain was sufficient to drive bacterial beta diversity between three different hyporheic flow path locations. Habitat heterogeneity was assessed by measuring soluble reactive phosphorous, nitrate, dissolved organic carbon, dissolved oxygen, and soluble total nitrogen levels seasonally at surface water infiltration, advection, and exfiltration zones. Significant spatial differences were detected in dissolved oxygen and nitrate levels, and seasonal differences were detected in dissolved oxygen, nitrate, and dissolved organic carbon levels. Denaturing gradient gel electrophoresis (DGGE) and cell counts indicated that bacterial diversity increased with abundance, and DGGE fingerprints covaried with nitrate levels where water infiltrated the hyporheic zone. The ribosomal gene phylogeny revealed that hyporheic habitat heterogeneity was sufficient to drive beta diversity between bacterial assemblages. Phylogenetic (P) tests detected sequence disparity between the flow path locations. Small distinct lineages of Firmicutes, Actinomycetes, Planctomycetes, and Acidobacteria defined the infiltration zone and alpha- and beta-proteobacterial lineages delineated the exfiltration and advection zone communities. These data suggest that spatial habitat heterogeneity drives hyporheic microbial community development and that attempts to understand functional differences between bacteria inhabiting nutritionally heterogeneous hyporheic environments might begin by focusing on the biology of these taxa.

Brain mapping of deception and truth telling about an ecologically valid situation: functional MR imaging and polygraph investigation--initial experience.

PURPOSE: To examine the neural correlates during deception and truth telling by using a functional magnetic resonance (MR) imaging technique and an ecologically valid task and to compare the results with those of a standard polygraph examination. MATERIALS AND METHODS: All subjects gave written informed consent for this HIPAA-approved study, which was approved by the institutional review board of Drexel University. Eleven healthy subjects (five female and six male subjects; mean age, 28.9 years) were randomly assigned to the group of guilty subjects or the group of nonguilty subjects. Each group consisted of two separate functional MR imaging conditions: "lie-only condition" and "truth-only condition." The lie-only condition was used to compare brain activity during a known lie to control questions and a subjective lie to relevant questions. The truth-only condition was used to compare brain activity during a known truthful response to control questions and a subjective truthful response to relevant questions. Functional MR images were acquired with an echo-planar sequence, and statistical analysis was performed. Physiologic responses were measured with a standard four-channel polygraph instrument. RESULTS: During the deception process, specific areas of the frontal lobe (left medial and left inferior frontal lobes), temporal lobe (right hippocampus and right middle temporal gyrus), occipital lobe (left lingual gyrus), anterior cingulate, right fusiform gyrus, and right sublobar insula were significantly active. During the truth telling process, specific areas of the frontal (left subcallosal gyrus or lentiform nucleus) and temporal (left inferior temporal gyrus) lobes were significantly active. The polygraph examination revealed 92% accuracy in deceptive subjects and 70% accuracy in truthful subjects. CONCLUSION: Specific areas of the brain involved in deception or truth telling can be depicted with functional MR imaging.

Integrated zone comparison polygraph technique accuracy with scoring algorithms.

The Integrated Zone Comparison Technique (IZCT) was utilized with computerized polygraph instrumentation as part of a blind study in the detection of deception. Three scoring algorithms: ASIT Poly Suite (Academy for Scientific Investigative Training's Horizontal Scoring and Algorithm for Chart Interpretation), PolyScore 5.5, and the Objective Scoring System (OSS) were assessed in the interpretation of the charts generated. Where "Inconclusives" were excluded, accuracy for the IZCT with all three algorithms was 100%. When "Inconclusives" were counted as errors, overall accuracy for the IZCT with ASIT Poly Suite was 90% and accuracy with PolyScore and the Objective Scoring System was 72%.

Relationship between communities and processes; new insights from a field study of a contaminated ecosystem.

We used a 93-year-old mine waste contamination gradient in alluvial soil to explore the relationship between ecosystem level functioning and community structure in a chronically stressed ecosystem. The sensitivity of broad functional parameters (in situ soil respiration, microbial biomass, above and below ground plant biomass) and microbial diversity [phospholipid fatty acid (PLFA) abundance and richness] were compared. Functional responses were linear with respect to contaminants while thresholds were detected in the community structural response to contamination along the gradient. For example, in situ soil respiration was negatively and linearly correlated to contamination concentration (R = -0.783, P < 0.01), but changes in microbial community structure only became evident where contaminant concentrations were greater than 28 times above background levels. Our results suggest that functional redundancy does not prevent depression of ecosystem function in the long-term.