Utilizing Malachite Green Derivatives to Diversify Fluorogen-Activating Proteins (FAPs)’ Applications

The field of cell imaging has been greatly advanced through the use of genetically encoded fluorescent probes. By genetically expressing to proteins of interest, fluorescent proteins have enabled noninvasively imaging of live cells to study protein-protein interaction, protein trafficking and protein expression and localization. In our group, we are dedicated to the development of protein reporters (FAPs) that can activate fluorescence from originally darkmolecules (fluorogens). Among these fluoromodules, FAPdL5** is a 25 kDa binder for malachite green (MG) derivatives that functions throughout living cells with thousands fold fluorescence activation and a low-picomolar dissociation constant. With rational designed MG derivatives, distinct applications have been found in single molecule imaging, physiological pH measurementsand protein detection as recombinant affinity probes. However, in previous applications, developments of functionalized MG derivatives were focused on energy-transfer pairs which a donor that signals the environmental change is usually covalently linked to MG molecule. Littlemodification has been done with MG chromophore itself.In this thesis, we explored the effects brought by introduction of different modifications on MG. First, by varying the charge of the linker, we have proposed and synthesized a series of MG derivatives with different net charge that directly alters the cell permeability. Secondly, a series of fluorinated MG derivatives were synthesized to investigate fluorination effect on spectra and affinity of MG-dL5**. Thirdly, a number of MG analogues with distinct absorption spectra were made to serve the purpose of multi-color imaging. Finally, a heavy-atom substituted MG (MG-2I)was made, which greatly increases singlet oxygen quantum yield upon binding to dL5**. This MG-2I/dL5** complex enables protein inactivation and targeted cell killing and rapid targeted lineage ablation in living larval and adult zebrafish

Cavity Shaving plus Lumpectomy versus Lumpectomy Alone for Patients with Breast Cancer Undergoing Breast-Conserving Surgery: A Systematic Review and Meta-Analysis

<div><p>The margin status is a well-established prognostic predictor for patients undergoing breast-conserving surgery (BCS). Recent data suggested that cavity shaving in addition to lumpectomy might be a promising approach for improving the clinical outcomes. We aimed to compare the efficacy and safety between cavity shaving plus lumpectomy and lumpectomy alone with a systematic review and meta-analysis. We searched the PubMed, Embase, and Cochrane CENTRAL databases for studies comparing cavity shaving with lumpectomy before June 10, 2016. Both comparative studies and self-control studies were included. A random-effects model was used to estimate the odds ratios (ORs) for positive margin rate, reoperation rate, recurrence rate, and weighted mean difference (WMD) for excised tissue volume. Twenty-six studies were included in the meta-analysis. The cavity shaving group had a significantly lower positive margin rate than the BCS-alone group (16.4% vs. 31.9%; OR = 0.41, 95% CI 0.32–0.53, P < 0.05). Cavity shaving was associated with a significantly decreased rate of reoperation (OR = 0.42, 95% CI 0.30–0.59, P < 0.05). The overall locoregional rate was low for cavity shaving and BCS-alone (3% vs. 4%). Cavity shaving had no significant effect on the risk of locoregional recurrence (OR = 0.86, 95% CI 0.32–2.35; P = 0.78). The excised tissue volume did not differ substantially between cavity shaving and BCS alone (WMD = −23.88, 95% CI −55.20 to 7.44, P = 0.14). For patients undergoing BCS, additional cavity shaving was an effective method to decrease the positive margin rate and avoid reoperation. The addition of cavity shaving did not appear to have excessive excised tissue volume compared with partial mastectomy alone.</p></div

Characteristics of the Included Studies.

Characteristics of the Included Studies.</p

The Publication Bias Shown by Funnel Plots.

<p>(A) Funnel plot for studies comparing the positive margin rate; (B) Funnel plot for studies comparing the reoperation rate.</p

Subcellular distribution of the test samples.

<p>Subcellular distribution of the test samples.</p

Forest Plot Comparing the Locoregional Recurrence Rate for Lumpectomy With and Without Cavity Shaving.

<p>Forest Plot Comparing the Locoregional Recurrence Rate for Lumpectomy With and Without Cavity Shaving.</p

Forest Plot Comparing the Positive Margin Rate for Partial Mastectomy With and Without Cavity Shaving.

<p>Forest Plot Comparing the Positive Margin Rate for Partial Mastectomy With and Without Cavity Shaving.</p

The experimental results (mean) on human protein data sets for investigating the usefulness of the correlations among the locations.

<p>The experimental results (mean) on human protein data sets for investigating the usefulness of the correlations among the locations.</p

Flow Diagram Showing the Study Selection Process.

<p>Flow Diagram Showing the Study Selection Process.</p