Non-Invasive Imaging of Cysteine Cathepsin Activity in Solid Tumors Using a 64Cu-Labeled Activity-Based Probe

The papain family of cysteine cathepsins are actively involved in multiple stages of tumorigenesis. Because elevated cathepsin activity can be found in many types of human cancers, they are promising biomarkers that can be used to target radiological contrast agents for tumor detection. However, currently there are no radiological imaging agents available for these important molecular targets. We report here the development of positron emission tomography (PET) radionuclide-labeled probes that target the cysteine cathepsins by formation of an enzyme activity-dependent bond with the active site cysteine. These probes contain an acyloxymethyl ketone (AOMK) functional group that irreversibly labels the active site cysteine of papain family proteases attached to a 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA) tag for labeling with 64Cu for PET imaging studies. We performed biodistribution and microPET imaging studies in nude mice bearing subcutaneous tumors expressing various levels of cysteine cathepsin activity and found that the extent of probe uptake by tumors correlated with overall protease activity as measured by biochemical methods. Furthermore, probe signals could be reduced by pre-treatment with a general cathepsin inhibitor. We also found that inclusion of a Cy5 tag on the probe increased tumor uptake relative to probes lacking this fluorogenic dye. Overall, these results demonstrate that small molecule activity-based probes carrying radio-tracers can be used to image protease activity in living subjects

An Optimized, Chemically Regulated Gene Expression System for Chlamydomonas

BACKGROUND: Chlamydomonas reinhardtii is a model system for algal and cell biology and is used for biotechnological applications, such as molecular farming or biological hydrogen production. The Chlamydomonas metal-responsive CYC6 promoter is repressed by copper and induced by nickel ions. However, induction by nickel is weak in some strains, poorly reversible by chelating agents like EDTA, and causes, at high concentrations, toxicity side effects on Chlamydomonas growth. Removal of these bottlenecks will encourage the wide use of this promoter as a chemically regulated gene expression system. METHODOLOGY: Using a codon-optimized Renilla luciferase as a reporter gene, we explored several strategies to improve the strength and reversibility of CYC6 promoter induction. Use of the first intron of the RBCS2 gene or of a modified TAP medium increases the strength of CYC6 induction up to 20-fold. In the modified medium, induction is also obtained after addition of specific copper chelators, like TETA. At low concentrations (up to 10 microM) TETA is a more efficient inducer than Ni, which becomes a very efficient inducer at higher concentrations (50 microM). Neither TETA nor Ni show toxicity effects at the concentrations used. Unlike induction by Ni, induction by TETA is completely reversible by micromolar copper concentrations, thus resulting in a transient "wave" in luciferase activity, which can be repeated in subsequent growth cycles. CONCLUSIONS: We have worked out a chemically regulated gene expression system that can be finely tuned to produce temporally controlled "waves" in gene expression. The use of cassettes containing the CYC6 promoter, and of modified growth media, is a reliable and economically sustainable system for the temporally controlled expression of foreign genes in Chlamydomonas

General self-efficacy and the effect of hospital workplace violence on doctors’ stress and job satisfaction in China

Objectives: This study aims at exploring associations of general self-efficacy (GSE), workplace violence and doctors' work-related attitudes. Material and Methods: In this study a cross-sectional survey design was applied. Questionnaires were administrated to 758 doctors working in 9 hospitals of Zhengzhou, Henan province, China, between June and October 2010. General information on age, gender, and years of working was collected, and the doctors' experience and witnessing workplace violence, job satisfaction, job initiative, occupational stress as well as GSE were measured. General linear regression analysis was performed in association analyses. Results: Both experiencing and witnessing workplace violence were significantly positively correlated with the level of occupational stress but significantly negatively correlated with job satisfaction, job initiative, and GSE. General self-efficacy significantly modified relationships between both experiencing and witnessing workplace violence with occupational stress (β = 0.49 for experiencing violence; β = 0.43 for witnessing violence; p 0.05). The levels of occupational stress declined significantly with the increase of GSE, while job satisfaction increased significantly along with its increase. The effects of GSE on occupational stress and job satisfaction weakened as the frequency of violence increased. Conclusions: The findings suggest that GSE can modify effects of workplace violence on health care workers' stress and job satisfaction. Enhancing GSE in combination with stress reduction may lead to facilitating health care workers' recovery from workplace violence, and thereby improving their work-related attitudes

Parameters for the mathematical modelling of Clostridium difficile acquisition and transmission: a systematic review

INTRODUCTION: Mathematical modelling of Clostridium difficile infection dynamics could contribute to the optimisation of strategies for its prevention and control. The objective of this systematic review was to summarise the available literature specifically identifying the quantitative parameters required for a compartmental mathematical model of Clostridium difficile transmission. METHODS: Six electronic healthcare databases were searched and all screening, data extraction and study quality assessments were undertaken in duplicate. Results were synthesised using a narrative approach. RESULTS: Fifty-four studies met the inclusion criteria. Reproduction numbers for hospital based epidemics were described in two studies with a range from 0.55 to 7. Two studies provided consistent data on incubation periods. For 62% of cases, symptoms occurred in less than 4 weeks (3-28 days) after infection. Evidence on contact patterns was identified in four studies but with limited data reported for populating a mathematical model. Two studies, including one without clinically apparent donor-recipient pairs, provided information on serial intervals for household or ward contacts, showing transmission intervals of <1 week in ward based contacts compared to up to 2 months for household contacts. Eight studies reported recovery rates of between 75%-100% for patients who had been treated with either metronidazole or vancomycin. Forty-nine studies gave recurrence rates of between 3% and 49% but were limited by varying definitions of recurrence. No study was found which specifically reported force of infection or net reproduction numbers. CONCLUSIONS: There is currently scant literature overtly citing estimates of the parameters required to inform the quantitative modelling of Clostridium difficile transmission. Further high quality studies to investigate transmission parameters are required, including through review of published epidemiological studies where these quantitative estimates may not have been explicitly estimated, but that nonetheless contain the relevant data to allow their calculation

The Study of Irregular Migration

AbstractThe study of irregular migration as a specific social phenomenon took off during the 70s in the US. Since then, the academic interest has continually grown and spread, first to Europe and, in the last years, to other regions worldwide. This interest can certainly be related to the increasing attention paid to the study of migrations more in general (Castles & Miller, 1993). The trend can be linked to those broad and complex social and economic changes, often subsumed under the concept of globalization. The specific focus on irregular migration, though gaining momentum throughout the 1980s, reached preeminent attention in the 1990s. On both sides of the Atlantic, the explosion of the so-called "migration crisis" (Zolberg & Benda, 2001) and the emergence of irregular migration as a widespread social fact raised the attention of public opinion and academics alike. Moreover, in recent years, what seemed at first to be an issue concerning only the high-income regions of the planet, now involves also medium and low-income ones, making irregular migration a truly global structural phenomenon (Cvajner & Sciortino, 2010a; Düvell, 2006)