Patient-to-patient spread of a single strain of Corynebacterium striatum causing infections in a surgical intensive care unit

Over a 12-month period, Corynebacterium striatum strains were isolated from clinical specimens from 14 patients admitted to a surgical intensive care unit. These isolates were identical by morphology and biotype and displayed the same antibiogram. Ten isolates were found to be the sole possible pathogen. These 10 isolates were from six patients, three of whom had signs of infection at the time of positive culture. Further typing was performed by random amplification of polymorphic DNA analysis, by which all strains were identical and were found to differ to various degrees from reference strains and from isolates found in clinical samples from other wards. In a case-control study the only independent risk factor for acquiring the strain was intubation for longer than 24 h (odds ratio, 20.09; 95

Laboratory-based surveillance in the molecular era: The typened model, a joint data-sharing platform for clinical and public health laboratories

Laboratory-based surveillance, one of the pillars of monitoring infectious disease trends, relies on data produced in clinical and/or public health laboratories. Currently, diagnostic laboratories worldwide submit strains or samples to a relatively small number of reference laboratories for characterisation and typing. However, with the introduction of molecular diagnostic methods and sequencing in most of the larger diagnostic and university hospital centres in high-income countries, the distinction between diagnostic and reference/public health laboratory functions has become less clear-cut. Given these developments, new ways of networking and data sharing are needed. Assuming that clinical and public health laboratories may be able to use the same data for their own purposes when sequence-based testing and typing are used, we explored ways to develop a collaborative approach and a jointly owned database (TYPENED) in the Netherlands. The rationale was that sequence data - whether produced to support clinical care or for surveillance -can be aggregated to meet both needs. Here we describe the development of the TYPENED approach and supporting infrastructure, and the implementation of a pilot laboratory network sharing enterovirus sequences and metadata

Cosmological Applications of Gravitational Lensing

The last decade has seen an enormous increase of activity in the field of gravitational lensing, mainly driven by improvements of observational capabilities. I will review the basics of gravitational lens theory, just enough to understand the rest of this contribution, and will then concentrate on several of the main applications in cosmology. Cluster lensing, and weak lensing, will constitute the main part of this review.Comment: 26 pages, including 2 figures (a third figure can be obtained from the author by request) gziped and uuencoded postscript file; to be published in Proceedings of the Laredo Advanced Summer School, Sept. 9

Pharmacodynamic Monitoring of Calcineurin Inhibition Therapy: Principles, Performance, and Perspectives

The calcineurin inhibitors (CNIs) cyclosporin A and tacrolimus are immunosuppressive drugs used extensively in allograft recipients. These drugs show large interindividual pharmacokinetic variation and are associated with severe adverse affects, including nephrotoxicity and cardiovascular disease. In current practice, CNIs are combined with other immunosuppressive drugs such as steroids and mycophenolate mofetil. Dosage is titrated based oil blood concentration measurement. For further optimization of calcineurin (CN) inhibition therapy, new monitoring strategies are required. Pharmacodynamic-monitoring strategies constitute novel approaches for optimization of CNIs therapy. This review focuses on the general aspects Of immunosuppressive drug pharmacodynamic monitoring and describes the methodologies used for monitoring CN inhibition therapy. Two different types of pharmacodynamic-monitoring strategies can be distinguished: (1) enzymatic strategies, which monitor inhibition of drug-target enzyme activity, and (2) immunologic strategies, which measure cellular responsiveness after in vitro simulated immunologic responses. Enzymatic tests are drug type-specific markers in which CN activity is directly determined. Immunologic strategies measure immune responsiveness at several levels, such as mRNA transcripts (intracellular) concentrations/ excretion of cytokines, expression of surface activation markers, and cell proliferation. This review also discusses analytical issues and clinical experience with these techniques. The call for new methodologies to evaluate immunosuppressive therapy has led to the development of a large variety of pharmacodynamic-monitoring strategies. The first reports of their clinical relevance are available, but further understanding of the analytical and clinical variables involved are required for the development of accurate, reproducible, and clinically relevant markers

Pharmacodynamic Monitoring of Calcineurin Inhibition Therapy: Principles, Performance, and Perspectives

The calcineurin inhibitors (CNIs) cyclosporin A and tacrolimus are immunosuppressive drugs used extensively in allograft recipients. These drugs show large interindividual pharmacokinetic variation and are associated with severe adverse affects, including nephrotoxicity and cardiovascular disease. In current practice, CNIs are combined with other immunosuppressive drugs such as steroids and mycophenolate mofetil. Dosage is titrated based oil blood concentration measurement. For further optimization of calcineurin (CN) inhibition therapy, new monitoring strategies are required. Pharmacodynamic-monitoring strategies constitute novel approaches for optimization of CNIs therapy. This review focuses on the general aspects Of immunosuppressive drug pharmacodynamic monitoring and describes the methodologies used for monitoring CN inhibition therapy. Two different types of pharmacodynamic-monitoring strategies can be distinguished: (1) enzymatic strategies, which monitor inhibition of drug-target enzyme activity, and (2) immunologic strategies, which measure cellular responsiveness after in vitro simulated immunologic responses. Enzymatic tests are drug type-specific markers in which CN activity is directly determined. Immunologic strategies measure immune responsiveness at several levels, such as mRNA transcripts (intracellular) concentrations/ excretion of cytokines, expression of surface activation markers, and cell proliferation. This review also discusses analytical issues and clinical experience with these techniques. The call for new methodologies to evaluate immunosuppressive therapy has led to the development of a large variety of pharmacodynamic-monitoring strategies. The first reports of their clinical relevance are available, but further understanding of the analytical and clinical variables involved are required for the development of accurate, reproducible, and clinically relevant markers.Nephrolog

The location of coat protein and viral RNAs of alfalfa mosaic virus in infected tobacco leaves and protoplasts

The location of coat protein of alfalfa mosaic virus (AIMV) strain 425 was determined in protoplasts isolated from infected tobacco leaves and in in vitro inoculated tobacco protoplasts, using immunocytochemistry on ultrathin frozen sections labeled with colloidal gold. In infected tobacco leaves 5 days postinoculation (p.i.) coat protein is present in the cytoplasm and nucleus, especially around the nucleolus. In in vitro inoculated tobacco protoplasts coat protein was not present in the nucleus 6 hr p.i. These results indicate that the presence of coat protein in the nucleus is not necessary for viral replication. However, coat protein could be detected in the nucleus 48 hr p.i. Probably coat protein or virus particles accumulate in the nucleus late in infection. Minus-strand RNA, as part of the replication complex, could be detected in a 650 g pellet fraction of infected tobacco leaves but not in the nucleus, suggesting that replication of AIMV occurs outside the nucleus

Extraction of collagen from fish skins and its use in manufacture of biopolymer films

The aim of this study was to extract collagen from fish skins and investigate the physical properties of the biodegradable films formed from the extracted fish collagen. Extraction of collagen using hydrogen peroxide or enzymatic methods proved to be unsuccessful. A white collagen substance was successfully extracted using a hydrochloric acid extraction method; however, it was unsuitable for the formation of edible films. An acetic acid extraction process successfully yielded a collagen substance, which could be used to form biodegradable collagen films. Differences in the mechanical film properties, tensile strength, Young's modulus, elongation and water vapor permeability were found between collagen films from different fish species. Films formed from collagen of New Zealand (NZ) hoki and NZ ling had greater elongation (%EL), tensile strength (TS) and elasticity in comparison with similar films from Irish (IRL) fish species. The addition of plasticizer to the collagen films had an effect on the mechanical properties of the film

A spectrophotometric assay for routine measurement of mammalian target of rapamycin activity in cell lysates

The mammalian target of rapamycin (mTOR) is an important mediator in the PI3K/AKT signaling pathway. mTOR is the target of immunosuppressive drugs, such as rapamycin and everolimus, that are used in transplant patients but also for the treatment of various cancers. We have developed a method for mTOR activity measurement in cell lysates that measures the phosphorylation of p70 S6 kinase by an enzyme linked immunosorbent assay (ELISA) protocol. Using an optimized lysis composition, activity could be measured in the peripheral blood mononuclear cells (PBMCs) isolated from blood. For the PBMCs, intra- and interassay variations of 7 and 10%, respectively, were found using one lot number of the kit. With different lot numbers, the interassay variation increased up to 21%. Activity remained constant for PBMC pool samples on storage for a period of more than 7 months. Activity could also be measured in CD3+ T-cells isolated from blood. In vitro experiments revealed maximum mTOR inhibition of 30% in PBMCs and 44% in T-cells. The in vitro inhibition in PBMCs could also be demonstrated by Western blotting. The mTOR activity measurements may be used to show in vivo inhibition in renal allograft patients during everolimus treatment and to study mTOR activity in various (tumor) cell types. (C) 2010 Elsevier Inc. All rights reserved.Nephrolog