Carboxypeptidase O is a Lipid Droplet-associated Enzyme Able to Cleave both Acidic and Polar C-terminal Amino Acids

Carboxypeptidase O (CPO) is a member of the M14 family of metallocarboxypeptidases with a preference for the cleavage of C-terminal acidic amino acids. CPO is largely expressed in the small intestine, although it has been detected in other tissues such as the brain and ovaries. CPO does not contain a prodomain, nor is it strongly regulated by pH, and hence appears to exist as a constitutively active enzyme. The goal of this study was to investigate the intracellular distribution and activity of CPO in order to predict physiological substrates and function. The distribution of CPO, when expressed in MDCK cells, was analyzed by immunofluorescence microscopy. Soon after addition of nutrient-rich media, CPO was found to associate with lipid droplets, causing an increase in lipid droplet quantity. As media became depleted, CPO moved to a broader ER distribution, no longer impacting lipid droplet numbers. Membrane cholesterol levels played a role in the distribution and in vitro enzymatic activity of CPO, with cholesterol enrichment leading to decreased lipid droplet association and enzymatic activity. The ability of CPO to cleave C-terminal amino acids within the early secretory pathway (in vivo) was examined using Gaussia luciferase as a substrate, C-terminally tagged with variants of an ER retention signal. While no effect of cholesterol was observed, these data show that CPO does function as an active enzyme within the ER where it removes C-terminal glutamates and aspartates, as well as a number of polar amino acids

Exosomes: From potential culprits to new therapeutic promise in the setting of cardiac fibrosis

Fibrosis is a significant global health problem associated with many inflammatory and degenerative diseases affecting multiple organs, individually or simultaneously. Fibrosis develops when extracellular matrix (ECM) remodeling becomes excessive or uncontrolled and is associated with nearly all forms of heart disease. Cardiac fibroblasts and myofibroblasts are the main effectors of ECM deposition and scar formation. The heart is a complex multicellular organ, where the various resident cell types communicate between themselves and with cells of the blood and immune systems. Exosomes, which are small extracellular vesicles, (EVs), contribute to cell-to-cell communication and their pathophysiological relevance and therapeutic potential is emerging. Here, we will critically review the role of endogenous exosomes as possible fibrosis mediators and discuss the possibility of using stem cell-derived and/or engineered exosomes as anti-fibrotic agents

Use of Colour Duplex Ultrasound as a First Line Surveillance Tool Following EVAR is Associated with a Reduction in Cost Without Compromising Accuracy

CT scanning remains the postoperative surveillance imaging modality of choice following EVAR. Concerns regarding cost, exposure to ionising radiation and intravenous contrast have led to a search for a less expensive, equally efficacious and safer method of monitoring EVAR patients after endograft deployment. This study evaluated the cost saving obtained if CDUS was employed as a first line surveillance tool following EVAR, as well as comparing the two entities in terms of efficacy. Patients & methods: Postoperative surveillance CTs and CDUS scans in the 145 patients who have undergone EVAR from 1st June 2003 to 1st July 2010 were compared for the detection of endoleak and determination of residual sac size. Results: Adopting a protocol where CDUS was employed as the first line surveillance tool following EVAR would result in a reduction in the number of postoperative CTs required in 2010 from 235 to 36. Based on 2010 costings, this would equate to an estimated reduction in expenditure from V117,500 to V34,915 a saving of V82,585. CDUS had a sensitivity of 100% and a specificity of 85% in the detection of endoleaks compared to CT. The positive predictive value was 28% and negative predictive value 100%. The Pearson Coefficient correlation of 0.96 indicates a large degree of correlation between CDUS and CT when measuring residual aneurysm size following EVAR. Conclusion: CDUS can replace CT as the first line surveillance tool following EVAR. This is associated with a significant reduction in the cost of surveillance without any loss of imaging accurac

Use of Colour Duplex Ultrasound as a First Line Surveillance Tool Following EVAR is Associated with a Reduction in Cost Without Compromising Accuracy

CT scanning remains the postoperative surveillance imaging modality of choice following EVAR. Concerns regarding cost, exposure to ionising radiation and intravenous contrast have led to a search for a less expensive, equally efficacious and safer method of monitoring EVAR patients after endograft deployment. This study evaluated the cost saving obtained if CDUS was employed as a first line surveillance tool following EVAR, as well as comparing the two entities in terms of efficacy. Patients & methods: Postoperative surveillance CTs and CDUS scans in the 145 patients who have undergone EVAR from 1st June 2003 to 1st July 2010 were compared for the detection of endoleak and determination of residual sac size. Results: Adopting a protocol where CDUS was employed as the first line surveillance tool following EVAR would result in a reduction in the number of postoperative CTs required in 2010 from 235 to 36. Based on 2010 costings, this would equate to an estimated reduction in expenditure from V117,500 to V34,915 a saving of V82,585. CDUS had a sensitivity of 100% and a specificity of 85% in the detection of endoleaks compared to CT. The positive predictive value was 28% and negative predictive value 100%. The Pearson Coefficient correlation of 0.96 indicates a large degree of correlation between CDUS and CT when measuring residual aneurysm size following EVAR. Conclusion: CDUS can replace CT as the first line surveillance tool following EVAR. This is associated with a significant reduction in the cost of surveillance without any loss of imaging accurac

Influence of the annealing ambient on structural and optical properties of rare earth implanted GaN

GaN films were implanted with Er and Eu ions and rapid thermal annealing was performed at 1000, 1100 and 1200 ⁰C in vacuum, in flowing nitrogen gas or a mixture of NH₃ and N₂. Rutherford backscattering spectrometry in the channeling mode was used to study the evolution of damage introduction and recovery in the Ga sublattice and to monitor the rare earth profiles after annealing. The surface morphology of the samples was analyzed by scanning electron microscopy and the optical properties by room temperature cathodoluminescence (CL). Samples annealed in vacuum and N₂ already show the first signs of surface dissociation at 1000 ⁰C. At higher temperature, Ga droplets form, at the surface. However, samples annealed in NH₃+N₂ exhibit a very good recovery of the lattice along with a smooth surface. These samples also show the strongest CL intensity for the rare earth related emissions in the green (for Er) and red (for Eu). After annealing at 1200 ⁰C in NH₃+N₂ the Eu implanted sample reveals the channeling qualities of an unimplanted sample and a strong increase of CL intensity is observed

A Cluster of Legionnaires' Disease and Associated Pontiac Fever Morbidity in Office Workers, Dublin, June-July 2008

In June and July 2008, two office workers were admitted to a Dublin hospital with Legionnaires' disease. Investigations showed that cooling towers in the basement car park were the most likely source of infection. However, positive results from cooling tower samples by polymerase chain reaction (PCR) did not correlate with subsequent culture results. Also, many employees reported Pontiac fever-like morbidity following notification of the second case of Legionnaires' disease. In total, 54 employees attended their general practitioner or emergency department with symptoms of Legionnaires' disease or Pontiac fever. However, all laboratory tests for Legionnaires' disease or Pontiac fever were negative. In this investigation, email was used extensively for active case finding and provision of time information to employees and medical colleagues. We recommend clarification of the role of PCR in the diagnosis of legionellosis and also advocate for a specific laboratory test for the diagnosis of the milder form of legionellosis as in Pontiac fever

Genome sequences and comparative genomics of two Lactobacillus ruminis strains from the bovine and human intestinal tracts

peer-reviewedBackground: The genus Lactobacillus is characterized by an extraordinary degree of phenotypic and genotypic diversity, which recent genomic analyses have further highlighted. However, the choice of species for sequencing has been non-random and unequal in distribution, with only a single representative genome from the L. salivarius clade available to date. Furthermore, there is no data to facilitate a functional genomic analysis of motility in the lactobacilli, a trait that is restricted to the L. salivarius clade. Results: The 2.06 Mb genome of the bovine isolate Lactobacillus ruminis ATCC 27782 comprises a single circular chromosome, and has a G+C content of 44.4%. In silico analysis identified 1901 coding sequences, including genes for a pediocin-like bacteriocin, a single large exopolysaccharide-related cluster, two sortase enzymes, two CRISPR loci and numerous IS elements and pseudogenes. A cluster of genes related to a putative pilin was identified, and shown to be transcribed in vitro. A high quality draft assembly of the genome of a second L. ruminis strain, ATCC 25644 isolated from humans, suggested a slightly larger genome of 2.138 Mb, that exhibited a high degree of synteny with the ATCC 27782 genome. In contrast, comparative analysis of L. ruminis and L. salivarius identified a lack of long-range synteny between these closely related species. Comparison of the L. salivarius clade core proteins with those of nine other Lactobacillus species distributed across 4 major phylogenetic groups identified the set of shared proteins, and proteins unique to each group. Conclusions: The genome of L. ruminis provides a comparative tool for directing functional analyses of other members of the L. salivarius clade, and it increases understanding of the divergence of this distinct Lactobacillus lineage from other commensal lactobacilli. The genome sequence provides a definitive resource to facilitate investigation of the genetics, biochemistry and host interactions of these motile intestinal lactobacilli

Epistemic and social scripts in computer-supported collaborative learning

Collaborative learning in computer-supported learning environments typically means that learners work on tasks together, discussing their individual perspectives via text-based media or videoconferencing, and consequently acquire knowledge. Collaborative learning, however, is often sub-optimal with respect to how learners work on the concepts that are supposed to be learned and how learners interact with each other. One possibility to improve collaborative learning environments is to conceptualize epistemic scripts, which specify how learners work on a given task, and social scripts, which structure how learners interact with each other. In this contribution, two studies will be reported that investigated the effects of epistemic and social scripts in a text-based computer-supported learning environment and in a videoconferencing learning environment in order to foster the individual acquisition of knowledge. In each study the factors ‘epistemic script’ and ‘social script’ have been independently varied in a 2×2-factorial design. 182 university students of Educational Science participated in these two studies. Results of both studies show that social scripts can be substantially beneficial with respect to the individual acquisition of knowledge, whereas epistemic scripts apparently do not to lead to the expected effects

Soft exosuits increase walking speed and distance after stroke

Finalist for best poster abstract and presentatio