Multicomponent Synthesis of Diverse 1,4-Benzodiazepine Scaffolds

The 1,4-benzodiazepine (BDZ) scaffold is of particular interest in drug design due to a balanced ensemble of beneficial physicochemical properties including a semirigid and compact diazepine ring with spatial placements of several substituents, combined with low number of rotatable bonds, hydrogen bond donors and acceptors, and intermediate lipophilicity. As an alternative to traditional multistep sequential syntheses, we designed routes employing one-pot MCRs to accelerate access diverse BDZ scaffolds in two or three steps

Tunable Carbon-Dot-Based Dual-Emission Fluorescent Nanohybrids for Ratiometric Optical Thermometry in Living Cells

The use of carbon-dot-based dual-emission fluorescent nanohybrids (DEFNs) as versatile nanothermometry devices for spatially resolved temperature measurements in living cells is demonstrated. The carbon dots (CDs) are prepared in the organic phase and display tunable photoluminescence (PL) across a wide visible range by adjusting the excitation wavelengths and extend of N-doping. DEFNs are formed in a straightforward fashion from CDs (emitting blue PL) and gold nanoclusters (AuNCs, emitting red PL). The DEFNs display ideal single-excitation, dual-emission with two well-resolved, intensity-comparable fluorescence peaks, and function in optical thermometry with high reliability and accuracy by exploiting the temperature sensitivity of their fluorescence intensity ratio (blue/red). Furthermore, the DEFNs have been introduced into cells, exhibiting good biocompatibility, and have facilitated physiological temperature measurements in the range of 25–45 °C; the DEFNs can therefore function as “non-contact” tools for the accurate measurement of temperature and its gradient inside a living cell

Mycoplasma-associated multidrug resistance of hepatocarcinoma cells requires the interaction of P37 and Annexin A2

<div><p>Mycoplasma infection has been reported to be associated with cancer migration, invasion, epithelial-mesenchymal transition as well as the resistance to nucleoside analogues chemotherapeutic drugs. In this study, we found that the sensitivity of hepatocarcinoma cells to Cisplatin, Gemcitabine and Mitoxantrone was increased by mycoplasma elimination. Similar to the effect of anti-mycoplasma agent, interrupting the interaction between <i>Mycoplasma hyorhinis</i> membrane protein P37 and Annexin A2 of host cells using the N-terminal of ANXA2 polypeptide enhanced the sensitivity of HCC97L cells to Gemcitabine and Mitoxantrone. Meanwhile, we did not observe any changes in expression or distribution of multidrug resistance associated transporters, ATP-Binding Cassette protein B1, C1 and G2, on the removal of mycoplasma. These results suggest that mycoplasma induces a resistance to multiple drugs in hepatocarcinoma cells which required the interaction of P37 and Annexin A2. The pathway downstream this interaction needs to be explored.</p></div

The effect of A2PP/MXF on the sensitivity of HCC97L to chemotherapeutic drugs.

<p>(A, B and C) Cell viability of HCC97L cells treated using GEM alone, GEM with the presence 3μg/mL of MXF or GEM with with three different concentrations of A2PP, which were 40 μM, 120 μM and 160 μM respectively. (D) Cell viability of HCC97L cells treated using MX alone, MX with the presence of MXF or MX with 160 μM of A2PP. Statistical testing was performed by comparing the logIC₅₀ values by means of an extra-sum-of-squares <i>F</i> test. ****<i>P</i> < 0.0001, ***<i>P</i> < 0.001, **<i>P</i> < 0.01, *<i>P</i> < 0.05 as compared with each group. IC₅₀ values, <i>F</i> values, degrees of freedom (DFn, DFd) and P values were provided in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0184578#pone.0184578.s006" target="_blank">S6 Table</a>. (E) Cell viability of HCC97L cells treated with different concentrations of A2PP or vehicle DMSO for 72 h (×200; bar, 50 μm). Statistical significance was determined by using paired two-tailed student’s <i>t</i>-test. <i>P</i> values, <i>t</i>-values and degree of freedom were provided in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0184578#pone.0184578.s007" target="_blank">S7 Table</a>. Error bars indicate SD of a representative experiment out of three independent experiments performed in triplicate.</p

The expression and subcellular location changes of ABC transporter family proteins with or without MXF treatment.

<p>(A) Protein expressions of ABCB1, ABCC1 and ABCG2 in HCC-97L cells treated with MXF for 7 days compared with non-treated controls. Statistical significance was determined by using unpaired two-tailed student’s <i>t</i>-test. <i>P</i> values, <i>t</i>-values and degrees of freedom were provided in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0184578#pone.0184578.s008" target="_blank">S8 Table</a>. Error bars indicate SD of a representative experiment out of three independent experiments performed in triplicate. The subcellular locations of ABCB1 (B), ABC1 (C) and ABCG2 (D) in HCC-97L cells treated with MXF for 7 days or non-treated controls. ZO-1 was used to delimitate the membrane (×400; bar, 100 μm).</p

The cell viability of hepatocarcinoma cells treated with different chemotheraputic drugs alone or with the presence of anti-mycoplasma antibiotics.

<p>Cell viability of HCC97L (A/G/M and B/H/N), Hep3B (C/I/O and D/J/P) and PLC/PRF/5 cell (E/K/Q and F/L/R), which were treated with CDDP/GEM/MX with or without AZI/MXF at the indicated concentrations. Error bars indicate SD of a representative experiment out of three independent experiments performed in triplicate. Statistical testing was performed by comparing the logIC₅₀ values by means of an extra-sum-of-squares <i>F</i> test. ****<i>P</i> < 0.0001 as compared to the chemotherapeutic drug alone controls. IC₅₀ values, <i>F</i> values, degrees of freedom (DFn, DFd) and <i>P</i> values were provided in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0184578#pone.0184578.s005" target="_blank">S5 Table</a>.</p

Primers used for qPCR of mycoplasma detection.

<p>Primers used for qPCR of mycoplasma detection.</p

Agreement in segmented layer locations between the manual and EdgeSelect methods.

<p>Solid lines indicate the mean difference of the agreement between the two methods and the dashed lines indicate mean ± 2 standard deviations. The filled symbols are data points from patients and the open symbols are from normal subjects. ILM: inner limiting membrane; ISe: the inner-segment/ellipsoid interface; RPE: the retinal/retinal pigment epithelium interface; BM: the Bruch's membrane.</p

Development of a Semi-Automatic Segmentation Method for Retinal OCT Images Tested in Patients with Diabetic Macular Edema

<div><p>Purpose</p><p>To develop EdgeSelect, a semi-automatic method for the segmentation of retinal layers in spectral domain optical coherence tomography images, and to compare the segmentation results with a manual method.</p><p>Methods</p><p>SD-OCT (Heidelberg Spectralis) scans of 28 eyes (24 patients with diabetic macular edema and 4 normal subjects) were imported into a customized MATLAB application, and were manually segmented by three graders at the layers corresponding to the inner limiting membrane (ILM), the inner segment/ellipsoid interface (ISe), the retinal/retinal pigment epithelium interface (RPE), and the Bruch's membrane (BM). The scans were then segmented independently by the same graders using EdgeSelect, a semi-automated method allowing the graders to guide/correct the layer segmentation interactively. The inter-grader reproducibility and agreement in locating the layer positions between the manual and EdgeSelect methods were assessed and compared using the Wilcoxon signed rank test.</p><p>Results</p><p>The inter-grader reproducibility using the EdgeSelect method for retinal layers varied from 0.15 to 1.21 µm, smaller than those using the manual method (3.36–6.43 µm). The Wilcoxon test indicated the EdgeSelect method had significantly better reproducibility than the manual method. The agreement between the manual and EdgeSelect methods in locating retinal layers ranged from 0.08 to 1.32 µm. There were small differences between the two methods in locating the ILM (p = 0.012) and BM layers (p<0.001), but these were statistically indistinguishable in locating the ISe (p = 0.896) and RPE layers (p = 0.771).</p><p>Conclusions</p><p>The EdgeSelect method resulted in better reproducibility and good agreement with a manual method in a set of eyes of normal subjects and with retinal disease, suggesting that this approach is feasible for OCT image analysis in clinical trials.</p></div

Mean, standard deviation, and concordance correlation of the inter-grader reproducibility of the EdgeSelect and the Manual methods.

<p>Layers are: inner limiting membrane (ILM), inner segment/ellipsoid interface (ISe), retina/retinal pigment epithelium interface (RPE), and Bruch's membrane (BM).</p