Genome-wide bacterial toxicity screening uncovers the mechanisms of toxicity of a cationic polystyrene nanomaterial

By exploiting a genome-wide collection of bacterial single-gene deletion mutants, we have studied the toxicological pathways of a 60-nm cationic (amino-functionalized) polystyrene nanomaterial (PS-NH(2)) in bacterial cells. The IC(50) of commercially available 60 nm PS-NH(2) was determined to be 158 μg/mL, the IC(5) is 108 μg/mL, and the IC(90) is 190 μg/mL for the parent E. coli strain of the gene deletion library. Over 4000 single nonessential gene deletion mutants of Escherichia coli were screened for the growth phenotype of each strain in the presence and absence of PS-NH(2). This revealed that genes clusters in the lipopolysaccharide biosynthetic pathway, outer membrane transport channels, ubiquinone biosynthetic pathways, flagellar movement, and DNA repair systems are all important to how this organism responds to cationic nanomaterials. These results, coupled with those from confirmatory assays described herein, suggest that the primary mechanisms of toxicity of the 60-nm PS-NH(2) nanomaterial in E. coli are destabilization of the outer membrane and production of reactive oxygen species. The methodology reported herein should prove generally useful for identifying pathways that are involved in how cells respond to a broad range of nanomaterials and for determining the mechanisms of cellular toxicity of different types of nanomaterials.     

Absence of sterol biosynthesis in oyster tissue culture

An oyster heart tissue culture was used to investigate the ability of the American oyster (Crassostrea virginica) to synthesize sterols from [¹⁴C]acetate. Lipids were extracted from cultured oyster heart cells and no label was found in the purified fatty acid fraction, 90% of which was located in the saturated and monounsaturated fractions. Although these results cannot rule out the possibility of sterol synthesis in other oyster tissues, this and previous evidence are in support of the hypothesis that the American oyster is unable to synthesize sterols.     

Performance evaluation of canine-associated Bacteroidales assays in a multi-laboratory comparison study

The contribution of fecal pollution from dogs in urbanized areas can be significant and is an often underestimated problem. Microbial source tracking methods (MST) utilizing quantitative PCR of dog-associated gene sequences encoding 16S rRNA of Bacteroidales are a useful tool to estimate these contributions. However, data about the performance of available assays are scarce. The results of a multi-laboratory study testing two assays for the determination of dog-associated Bacteroidales (DogBact and BacCan-UCD) on 64 single and mixed fecal source samples created from pooled fecal samples collected in California are presented here. Standardization of qPCR data treatment lowered inter-laboratory variability of sensitivity and specificity results. Both assays exhibited 100% sensitivity. Normalization methods are presented that eliminated random and confirmed non-target responses. The combination of standardized qPCR data treatment, use of normalization via a non-target specific Bacteroidales assay (GenBac3), and application of threshold criteria improved the calculated specificity significantly for both assays. Such measures would reasonably improve MST data interpretation not only for canine-associated assays, but for all qPCR assays used in identifying and monitoring fecal pollution in the environment.     

Dysfunctional cGMP Signaling Leads to Age-Related Retinal Vascular Alterations and Astrocyte Remodeling in Mice

The nitric oxide–guanylyl cyclase-1–cyclic guanylate monophosphate (NO–GC-1–cGMP) pathway is integral to the control of vascular tone and morphology. Mice lacking the alpha catalytic domain of guanylate cyclase (GC1^−/−) develop retinal ganglion cell (RGC) degeneration with age, with only modest fluctuations in intraocular pressure (IOP). Increasing the bioavailability of cGMP in GC1^−/− mice prevents neurodegeneration independently of IOP, suggesting alternative mechanisms of retinal neurodegeneration. In continuation to these studies, we explored the hypothesis that dysfunctional cGMP signaling leads to changes in the neurovascular unit that may contribute to RGC degeneration. We assessed retinal vasculature and astrocyte morphology in young and aged GC1^−/− and wild type mice. GC1^−/− mice exhibit increased peripheral retinal vessel dilation and shorter retinal vessel branching with increasing age compared to Wt mice. Astrocyte cell morphology is aberrant, and glial fibrillary acidic protein (GFAP) density is increased in young and aged GC1^−/− mice, with areas of dense astrocyte matting around blood vessels. Our results suggest that proper cGMP signaling is essential to retinal vessel morphology with increasing age. Vascular changed are preceded by alterations in astrocyte morphology which may together contribute to retinal neurodegeneration and loss of visual acuity observed in GC1^−/− mice.     

Low energy ballasted flotation

A novel process which involves the replacement or supplementation of bubbles in the dissolved air flotation process with low density beads is presented. The work comprised a series of bench-scale flotation trials treating three commonly encountered algal species (Microcystis, Melosira and Chlorella) that were removed in a flotation cell configured as either: conventional dissolved air flotation (DAF); ballasted flotation using low density 70 μm glass beads with a density of 100 kg m⁻³; or a hybrid process of ballasted flotation combined with conventional DAF. Results indicated that the bead only system was capable of achieving better residual turbidity than standard DAF at bead concentrations of 500 mg L⁻¹. Addition of beads in combination with standard DAF reduced turbidity further to even lower residual turbidity levels. Algae removal was improved when glass beads were dosed, but removal was dependent on algal species. Microcystis was removed by 97% for bead only systems and this removal did not change significantly with the addition of air bubbles. Melosira was the next best removed algae with bead only dosed systems giving similar removals to that achieved by standard DAF using a 10% air recycle ratio (81 and 76% removal respectively). Chlorella was the least well removed algae by bead only systems (63% removal). However, removal was rapidly improved to 86% by the addition of air bubbles using only a 2% recycle ratio. Energy estimations suggested that at least a 50% energy reduction could be achieved using the process offering a potential route for future development of low energy separation processes for algae removal.     

A category adjustment approach to memory for spatial location in natural scenes.

Memories for spatial locations often show systematic errors toward the central value of the surrounding region. This bias has been explained using a Bayesian model in which fine-grained and categorical information are combined (Huttenlocher, Hedges, & Duncan, 1991). However, experiments testing this model have largely used locations contained in simple geometric shapes. Use of this paradigm raises 2 issues. First, do results generalize to the complex natural world? Second, what types of information might be used to segment complex spaces into constituent categories? Experiment 1 addressed the 1st question by showing a bias toward prototypical values in memory for spatial locations in complex natural scenes. Experiment 2 addressed the 2nd question by manipulating the availability of basic visual cues (using color negatives) or of semantic information about the scene (using inverted images). Error patterns suggest that both perceptual and conceptual information are involved in segmentation. The possible neurological foundations of location memory of this kind are discussed. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Serotonin 5-HT2 receptors in schizophrenia: a PET study using [18F]setoperone in neuroleptic-naive patients and normal subjects

BACKGROUND: In the FRISC trial, dalteparin 120 IU/kg body weight twice daily for unstable coronary artery disease was safe and reduced the risk of new coronary events. This risk reduction was maintained during the following extended treatment with a fixed dose of 7500 IU dalteparin once daily. METHODS AND RESULTS: Minor bleeding was more frequent in women compared with men: relative risk (CI) 2.88 (1.78 to 4.67) during the weight-adjusted and 2.36 (1.37 to 2.63) during the fixed dose treatment. The anti-Xa activity determined in samples (n = 175) obtained during the acute phase treatment was higher in women compared with men (P <.001) and in nonsmokers compared with smokers (<.001) in multiple regression analysis. Also, during the fixed-dose treatment (n = 131) an independent relation between anti-Xa activity and sex (P <.001), but not smoking habits, persisted. CONCLUSION: To improve future low-molecular-weight heparin dose regimens for the treatment of acute coronary syndromes, it might be important to consider the influence of sex and smoking habits.