Molecular cloning and biochemical characterization of Xaa-Pro dipeptidyl-peptidase from Streptococcus mutans and its inhibition by anti-human DPP IV drugs

Streptococcus mutans harbours an intracellular, human DPP IV-analogous enzyme Xaa-Pro dipeptidyl-peptidase (EC 3.4.14.11). According to previous reports, an extracellular isozyme in S. gordonii and S. suis has been associated with virulence. Speculating that even an intracellular form may aid in virulence of S. mutans, we have tried to purify, characterize and evaluate enzyme inhibition by specific inhibitors. The native enzyme was partially purified by ion-exchange and gel filtration chromatography. Owing to low yield, the enzyme was overexpressed in Lactococcus lactis and purified by affinity chromatography. The recombinant enzyme (rSm-XPDAP) had a specific activity of 1070 U mg-1, while the Vmax and Km were 7 μM min-1 and 89 ± 7 μM (n = 3), respectively. The serine protease inhibitor phenylmethylsulphonyl fluoride and a DPP IV-specific inhibitor diprotin A proved to be active against rSm-XPDAP. As a novel approach, the evaluation of the effect of anti-human DPP IV (AHD) drugs on rSm-XPDAP activity found saxagliptin to be effective to some extent (Ki = 129 ± 16 μM), which may lead to the synthesis and development of a new class of antimicrobial agents

A Review of Exposure Assessment Methods in Epidemiological Studies on Incinerators

Incineration is a common technology for waste disposal, and there is public concern for the health impact deriving from incinerators. Poor exposure assessment has been claimed as one of the main causes of inconsistency in the epidemiological literature. We reviewed 41 studies on incinerators published between 1984 and January 2013 and classified them on the basis of exposure assessment approach. Moreover, we performed a simulation study to explore how the different exposure metrics may influence the exposure levels used in epidemiological studies. 19 studies used linear distance as a measure of exposure to incinerators, 11 studies atmospheric dispersion models, and the remaining 11 studies a qualitative variable such as presence/absence of the source. All reviewed studies utilized residence as a proxy for population exposure, although residence location was evaluated with different precision (e.g., municipality, census block, or exact address). Only one study reconstructed temporal variability in exposure. Our simulation study showed a notable degree of exposure misclassification caused by the use of distance compared to dispersion modelling. We suggest that future studies (i) make full use of pollution dispersion models; (ii) localize population on a fine-scale; and (iii) explicitly account for the presence of potential environmental and socioeconomic confounding

The coherent dynamics of photoexcited green fluorescent proteins

The coherent dynamics of vibronic wave packets in the green fluorescent protein is reported. At room temperature the non-stationary dynamics following impulsive photoexcitation displays an oscillating optical transmissivity pattern with components at 67 fs (497 cm-1) and 59 fs (593 cm-1). Our results are complemented by ab initio calculations of the vibrational spectrum of the chromophore. This analysis shows the interplay between the dynamics of the aminoacidic structure and the electronic excitation in the primary optical events of green fluorescent proteins.Comment: accepted for publication in Physical Review Letter

Performance of the diamond active target prototype for the PADME experiment at the DAΦ\PhiNE BTF

The PADME experiment at the DA$\Phi$NE Beam-Test Facility (BTF) is designed to search for the gauge boson of a new $\rm U(1)$ interaction in the process e$^+$e$^-\rightarrow\gamma$+$\rm A'$, using the intense positron beam hitting a light target. The $\rm A'$, usually referred as dark photon, is assumed to decay into invisible particles of a secluded sector and it can be observed by searching for an anomalous peak in the spectrum of the missing mass measured in events with a single photon in the final state. The measurement requires the determination of the 4-momentum of the recoil photon, performed by a homogeneous, highly segmented BGO crystals calorimeter. A significant improvement of the missing mass resolution is possible using an active target capable to determine the average position of the positron bunch with a resolution of less than 1 mm. This report presents the performance of a real size $\rm (2x2 cm^2)$ PADME active target made of a thin (50 $\mu$m) diamond sensor, with graphitic strips produced via laser irradiation on both sides. The measurements are based on data collected in a beam test at the BTF in November 2015.Comment: 7 pages, 10 figure

Nanoscale Nonlinear Spectroscopy with Electron Beams

We theoretically demonstrate the ability of electron beams to probe the nonlinear photonic response with nanometer spatial resolution, well beyond the capabilities of existing optical techniques. Although the interaction of electron beams with photonic modes is generally weak, the use of optical pumping produces stimulated electron–light interactions that can exceed order-unity probabilities in photon-induded near field electron microscopy (PINEM). Here, we demonstrate that PINEM can locally and quantitatively probe the nonlinear optical response. Specifically, we predict a dependence of PINEM electron spectra on the sample nonlinearity that can reveal the second-harmonic (SH) response of optical materials with nanometer resolution, observed through asymmetries between electron energy losses and gains. We illustrate this concept by showing that PINEM spectra are sensitive to the SH near field of centrosymmetric structures and by finding substantial spectral asymmetries in geometries for which the linear interaction is reduced