A complementary role of multiparameter flow cytometry and high-throughput sequencing for minimal residual disease detection in chronic lymphocytic leukemia: an European Research Initiative on CLL study.

In chronic lymphocytic leukemia (CLL) the level of minimal residual disease (MRD) after therapy is an independent predictor of outcome. Given the increasing number of new agents being explored for CLL therapy, using MRD as a surrogate could greatly reduce the time necessary to assess their efficacy. In this European Research Initiative on CLL (ERIC) project we have identified and validated a flow-cytometric approach to reliably quantitate CLL cells to the level of 0.0010% (10(-5)). The assay comprises a core panel of six markers (i.e. CD19, CD20, CD5, CD43, CD79b and CD81) with a component specification independent of instrument and reagents, which can be locally re-validated using normal peripheral blood. This method is directly comparable to previous ERIC-designed assays and also provides a backbone for investigation of new markers. A parallel analysis of high-throughput sequencing using the ClonoSEQ assay showed good concordance with flow cytometry results at the 0.010% (10(-4)) level, the MRD threshold defined in the 2008 International Workshop on CLL guidelines, but it also provides good linearity to a detection limit of 1 in a million (10(-6)). The combination of both technologies would permit a highly sensitive approach to MRD detection while providing a reproducible and broadly accessible method to quantify residual disease and optimize treatment in CLL.

Smoking cessation after an acute coronary syndrome: immediate quitters are successful quitters.

BACKGROUND: Cardiovascular disease (CVD) prevention guidelines stress the importance of smoking cessation and recommend intensive follow-up. To guide the development of such cessation support strategies, we analysed the characteristics that are associated with successful smoking cessation after an acute coronary syndrome (ACS). METHODS: We used data from the Randomised Evaluation of Secondary Prevention for ACS patients coordinated by Outpatient Nurse SpEcialists (RESPONSE) trial (n = 754). This was designed to quantify the impact of a nurse-coordinated prevention program, focusing on healthy lifestyles, traditional CVD risk factors and medication adherence. For the current analysis we included all smokers (324/754, 43 %). Successful quitters were defined as those who reported abstinence at 1 year of follow-up. RESULTS: The majority of successful quitters quit immediately after the ACS event and remained abstinent through 1 year of follow-up, without extra support (128/156, 82 %). Higher education level (33 vs. 15 %, p < 0.01), no history of CVD (87 vs. 74 %, p < 0.01) and being on target for LDL-cholesterol level at 1 year (78 vs. 63 %, p < 0.01) were associated with successful quitting. CONCLUSION: The majority of successful quitters at 1 year stopped immediately after their ACS. Patients in this group showed that it was within their own ability to quit, and they did not relapse through 1 year of follow-up. Our study indicates that in a large group of patients who quit immediately after a life-threatening event, no relapse prevention program is needed.

Ultraviolet laser-induced fluorescence of human stomach tissues: detection of cancer tissues by imaging techniques.

BACKGROUND AND OBJECTIVE: The background for this work was several literature reports on applications of the fluorescence methods to detection and localization of human cancers. The objective of our study has been to investigate if such an approach could be applied for the detection of gastric cancers. STUDY DESIGN/MATERIALS AND METHODS: Our study was designed in such a way that spectrally resolved images of laser-induced fluorescence of human gastric mucosa were collected and assessed from a point of view of elaborating an algorithm allowing for a differentiation between malignant and premalignant lesions and areas of normal mucosa. The method involved exciting the autofluorescence with ultraviolet light (325 nm, He-Cd laser). The images were recorded in vitro in six regions of a visible spectrum using a cooled CCD camera. The material for study was 21 resected specimens for which altogether 72 surface areas were examined. RESULTS: The main result is the observation that a difference of the fluorescence intensities measured at 440 nm and 395 nm, both normalized to intensity measured at 590 nm, differs significantly for the tissues of interest. CONCLUSION: Using that difference as a diagnostic parameter, it was possible to classify malignant tumor tissues with a sensitivity of 96% and a predictive value of 42%, whereas the same approach applied to abnormal but not tumor stomach tissues gave values of 80%, and 98%, respectively.