Rapid antimicrobial susceptibility testing of urinary tract isolates and samples by flow cytometry

A multiparametric flow cytometry antimicrobial susceptibility test was developed and its performance was evaluated on clinical urine isolates and samples in comparison with standard methods. Alterations in cytoplasmic membrane integrity were monitored by propidium iodide, and the anionic probe bis-(1,3-dibutylbarbituric acid) trimethine oxonol (DiBAC4(3)) was used to measure changes in membrane potential. Microbial size and cellular content were analysed by light scattering. Twelve antibiotics were tested on 6 ATCC control strains, 22 urine isolates and 19 clinical urine samples, variously containing Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, Proteus mirabilis, Enterococcus faecalis, Staphylococcus aureus, S. saprophyticus and S. epidermidis. Agreement between the flow cytometry results, broth microdilution and disk diffusion tests was 93.9% (n = 328 tests). Of the 20 discrepancies observed, 18 were for species other than E. coli. Perfect correlation was obtained with five antibiotics, whereas norfloxacin, nitrofurantoin and tetracycline were responsible for 13(65%) of the 20 discrepancies.     

The <Emphasis Type="Italic">i</Emphasis><Superscript>2</Superscript><Emphasis Type="Italic">Snake</Emphasis> Robotic Platform for Endoscopic Surgery

Endoscopic procedures have transformed minimally invasive surgery as they allow the examination and intervention on a patient’s anatomy through natural orifices, without the need for external incisions. However, the complexity of anatomical pathways and the limited dexterity of existing instruments, limit such procedures mainly to diagnosis and biopsies. This paper proposes a new robotic platform: the Intuitive imaging sensing navigated and kinematically enhanced (\(i^{2}Snake\)) robot that aims to improve the field of endoscopic surgery. The proposed robotic platform includes a snake-like robotic endoscope equipped with a camera, a light-source and two robotic instruments, supported with a robotic arm for global positioning and for insertion of the \(i^{2}Snake,\) and a master interface for master–slave teleoperation. The proposed robotic platform design focuses on ergonomics and intuitive control. The control workflow was first validated in simulation and then implemented on the robotic platform. The results are consistent with the simulation and show the clear clinical potential of the system. Limitations such as tendon backlash and elongation over time will be further investigated by means of combined hardware and software solutions. In conclusion, the proposed system contributes to the field of endoscopic surgical robots and could allow to perform more complex endoscopic surgical procedures while reducing patient trauma and recovery time.     

Attempted Passive Prophylaxis with a Monoclonal Anti-Burkholderia Pseudomallei Exopolysaccharide Antibody in a Murine Model of Melioidosis

Melioidosis is a severe gram-negative infection caused by the facultative intracellular bacterium Burkholderia pseudomallei, which is responsible for a broad spectrum of symptoms in both humans and animals. No licensed vaccine currently exists. This study evaluated the protective effect of a monoclonal antibody (Mab Ps6F6) specific to B. pseudomallei exopolysaccharide in an outbred murine model of sub-acute melioidosis. When administered before the infectious challenge, Ps6F6 significantly increased resistance to infection and restrained bacterial burden in the spleen over a 30-days period. Patterns of IFN-γ production were similar in the treated and non treated groups of mice. However, Ps6F6 lowered IFN-γ levels over the duration of the assay period, except on day 1, suggesting a transient and rapid production of IFN-γ under Ps6F6 control. Minor but persisting increases occurred in IL-12 levels while TNF-α was detected only in the controls at the later stages of infection. No IL-10 secretion was detected in both groups of mice. These data suggest that passive prophylaxis with Mab Ps6F6 provide a moderate and transient induction of inflammatory responses in infected mice but failed to trigger a sterilizing protective immunity.     

N1pc reversal following repeated eccentric visual stimulation

Early event‐related potential (ERP) hemispheric asymmetries recorded at occipitoparietal sites are usually observed following the sudden onset of a lateral peripheral stimulus. This is usually reflected in an onset‐locked larger N1 over the posterior contralateral hemisphere relative to the ipsilateral hemisphere, an early ERP asymmetry labeled N1pc. When the peripheral sudden onset is followed by a central stimulus, or by a bilaterally balanced visual array of stimuli, these events evoke a reversed N1pc, that is, a larger N1 over the hemisphere ipsilateral to the peripheral sudden onset. This N1pc reversal has been taken as evidence for a remapping of the visual space from an absolute, retinally based frame of reference to a relative, attentionally based frame of reference that codes the spatial positions of objects relative to the peripheral sudden onset, rather than relative to the fovea. Here, we pit the reference frame‐remapping account against an alternative account based on reduced neural reactivity following the peripheral sudden onset. In three experiments, we varied the spatial location of an object relative to a preceding sudden onset, and tested the opposite predictions generated by the frame‐remapping and the reduced neural reactivity accounts. Taken together, the results from the present experiments were consistent with the reduced neural reactivity account and inconsistent with the frame‐remapping account.     

DOES VISUAL SUBORDINATE-LEVEL CATEGORISATION ENGAGE THE FUNCTIONALLY DEFINED FUSIFORM FACE AREA?

Functional magnetic resonance imaging was used to compare brain activation associated with basic-level (e.g. bird) and subordinate-level (e.g. eagle) processing for both visual and semantic judgements. We localised the putative face area for 11 subjects, who also performed visual matching judgements for pictures and aurally presented words. The middle fusiform and occipital gyri were recruited for subordinate minus basic visual judgements, reflecting additional perceptual processing. When the face area was localised individually for each subject, analyses in the middle fusiform gyri revealed that subordinate-level processing activated the individuals face area. We propose that what is unique about the way faces engage this region is the focal spatial distribution of the activation rather than the recruitment of the face per se. Eight subjects also performed semantic judgements on aurally presented basic- and subordinate-level words. The parahippocampal gyri were more activated for subordinate-level than basic-level semantic judgements. Finally, the left posterior inferior temporal gyrus was activated for subordinate-level judgements, both visual and semantic, as well as during passive viewing of faces.     

Functional proteomics mapping of a human signaling pathway

Access to the human genome facilitates extensive functional proteomics studies. Here, we present an integrated approach combining large-scale protein interaction mapping, exploration of the interaction network, and cellular functional assays performed on newly identified proteins involved in a human signaling pathway. As a proof of principle, we studied the Smad signaling system, which is regulated by members of the transforming growth factor beta (TGFbeta) superfamily. We used two-hybrid screening to map Smad signaling protein-protein interactions and to establish a network of 755 interactions, involving 591 proteins, 179 of which were poorly or not annotated. The exploration of such complex interaction databases is improved by the use of PIMRider, a dedicated navigation tool accessible through the Web. The biological meaning of this network is illustrated by the presence of 18 known Smad-associated proteins. Functional assays performed in mammalian cells including siRNA knock-down experiments identified eight novel proteins involved in Smad signaling, thus validating this integrated functional proteomics approach.     

The nicotinic acetylcholine receptor gene family of the honey bee, Apis mellifera

Nicotinic acetylcholine receptors (nAChRs) mediate fast cholinergic synaptic transmission and play roles in many cognitive processes. They are under intense research as potential targets of drugs used to treat neurodegenerative diseases and neurological disorders such as Alzheimer's disease and schizophrenia. Invertebrate nAChRs are targets of anthelmintics as well as a major group of insecticides, the neonicotinoids. The honey bee, Apis mellifera, is one of the most beneficial insects worldwide, playing an important role in crop pollination, and is also a valuable model system for studies on social interaction, sensory processing, learning, and memory. We have used the A. mellifera genome information to characterize the complete honey bee nAChR gene family. Comparison with the fruit fly Drosophila melanogaster and the malaria mosquito Anopheles gambiae shows that the honey bee possesses the largest family of insect nAChR subunits to date (11 members). As with Drosophila and Anopheles, alternative splicing of conserved exons increases receptor diversity. Also, we show that in one honey bee nAChR subunit, six adenosine residues are targeted for RNA A-to-I editing, two of which are evolutionarily conserved in Drosophila melanogaster and Heliothis virescens orthologs, and that the extent of editing increases as the honey bee lifecycle progresses, serving to maximize receptor diversity at the adult stage. These findings on Apis mellifera enhance our understanding of nAChR functional genomics and provide a useful basis for the development of improved insecticides that spare a major beneficial insect species.     

Initiation of pre-B cell receptor signaling: common and distinctive features in human and mouse

B cell development in the bone marrow is a highly regulated process and expression of a functional pre-BCR represents a crucial checkpoint, common to human and mouse. In this review, we discuss pre-BCR analogies and differences between the two species leading to pre-B cell differentiation and proliferation. In addition, the mechanisms triggering pre-BCR activation are reviewed, taking into account the recent report of heparan sulfates and galectin 1 as stromal cell-derived pre-BCR ligands. Finally, ligand-induced pre-BCR activation models are proposed on the bases of the differences reported for pre-BCR and IL7 dependencies in the two species.