We have identified a novel antisense ODN sequence (ODN 2009) that effectively reduces the viability of small-cell lung cancer cells by reducing Bcl-2 levels and facilitating apoptosis.

Background:Work with primary cells is inherently limited by source availability and life span in culture. Flow cytometry offers extensive analytical opportunities but generally requires high cell numbers for an experiment. Methods: We have developed assays on a microfluidic system, which allow flow cytometric analysis of apoptosis and protein expression with a minimum number of fluorescently stained primary cells. In this setup, the cells are moved by pressure-driven flow inside a network of microfluidic channels and are analyzed individually by twochannel fluorescence detection. For some assays the staining reactions can be performed on-chip and the analysis is done without further washing steps. Results:We have successfully applied the assays to evaluate (a) activation of E-selectin (CD62E) expression by

L ab-on-a-chip technology achieves a reduction of sample and reagent volume and automates complex laboratory processes. Here, we present the implementation of cell assays on a microfluidic platform using disposable microfluidic chips. The applications are based on the controlled movement of cells by pressure-driven flow inside networks of microfluidic channels. Cells are hydrodynamically focused and pass the fluorescence detector in single file. Initial applications are the determination of protein expression and apoptosis parameters. The microfluidic system allows unattended measurement of six samples per chip. Results obtained with the microfluidic chips showed good correlation with data obtained using a standard flow cytometer.

No abstract

No abstract

No abstract

e20652 Background: Pneumocystis pneumonia (PCP) is a frequent and serious infectious complication in tumor patients. PCP is caused by Pneumocystis jirovecii, a fungal pathogen which is uncultivable in vitro. Highly-sensitive and time-efficient diagnostic tools are needed for early PCP treatment, especially in patients with impaired immune status. Methods: We evaluated a prototype of a new non-invasive molecular diagnostic tool (Unyvero System) using polymerase chain reaction (PCR) for P. jirovecii detection in respiratory samples (bronchoalveolar lavage fluid, tracheal aspirates, sputum) of hospitalized patients suffering from pneumonia. Results were obtained in an open-label, multicentric, non-randomized clinical study (CS-2011) conducted in 5 European hospitals. Unyvero system test results were compared to study site specific PCR results for P. jirovecii detection. Discrepant results were analysed for clinical relevance of molecular diagnostic tool results. Results: P. jirovecii was detected in 11 patients (1.49 %) out of 739 evaluable study patients: Of these, 8 cases (1.08 %) were detected only with the Unyvero System, 2 cases only with site-established techniques and 1 case with both methods. An independent academic cross-check of these 8 Unyvero System -positive cases showed that 5 cases were false positives due to signal reading failure. The remaining 3 cases could be confirmed as true positives by control PCP-PCR, indicating Unyvero P. jiroveciidetection results of 2 false negatives, 4 true positives and 5 false positives. Conclusions: The overall detection rate for P. jirovecii was low (11 out of 739) in this study population. Although more positive cases were found using the Unyvero System, it remains unclear if this was of clinical significance, especially as two clinically confirmed PCP cases could not be detected. The Unyvero System prototypes had a rate of 0.7% false positive results (5 out of 739 samples) which might have led to an overtreatment under real conditions. However, the correct detection of 4 positive cases within 3 hours might render the Unyvero System a valuable diagnostic future tool.

No abstract