Protein fragment complementation assays (PCAs) based on different reporter proteins have been described as powerful tools for monitoring dynamic protein-protein interactions in living cells. The present study describes the construction of a PCA system based on genetic splitting of TEM-1 beta-lactamase for the selection of proteins specifically interacting in the periplasm of Escherichia coli bacterial cells, and its application for the selection of affibody molecules binding human tumour necrosis factor-alpha (TNF-alpha) from a combinatorial library. Vectors encoding individual members of a naïve 10(9) affibody protein library fused to a C-terminal fragment of the beta-lactamase reporter were distributed via phage infection to a culture of cells harbouring a common construct encoding a fusion protein between a non-membrane anchored version of a human TNF-alpha target and the N-terminal segment of the reporter.
View Article and Find Full Text PDFCell-free circulating DNA isolated from the plasma of individuals with cancer has been shown to harbor cancer-associated changes in DNA methylation, and thus it represents an attractive target for biomarker discovery. However, the reliable detection of DNA methylation changes in body fluids has proven to be technically challenging. Here we describe a novel combination of methods that allows quantitative and sensitive detection of DNA methylation in minute amounts of DNA present in body fluids (quantitative Methylation Analysis of Minute DNA amounts after whole Bisulfitome Amplification, qMAMBA).
View Article and Find Full Text PDFBackground: Approximately 10-20% of stroke cases have normal blood pressure (BP). The objective of this study was to explore whether the risk of ischemic stroke is related to the carotid intima-media thickness (CIMT) and atherosclerotic lesions in a cohort of subjects with normal BP.
Methods: Common CIMT and the presence of carotid plaque were determined by B-mode ultrasound in 6,103 subjects, randomly recruited between 1991 and 1994 from the 'Malmo Diet and Cancer' study.
Background: Few studies have focused on the quality of secondary prevention among long-term stroke survivors. This study explores the intensity of medical intervention and the impact of lifestyle and other risk factors on the long-term stroke prognosis in a population-based setting of Swedish individuals with a history of stroke.
Methods: A population-based cohort (n = 28,449) in Malmo, Sweden, was recruited during 1991-1996.
Objective: Breast cancers detected between screening examinations can influence the sensitivity of a screening programme. Studies of the prognosis of these so-called interval breast cancers show diverging results. We investigated the course of interval breast cancer over time in the Malmö Mammographic Screening Trial (MMST) 1976-86 and the Malmö Mammographic Service Screening Programme (MMSSP) 1990-99.
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