Dynamic Python-Based Method Provides Quantitative Analysis of Intercellular Junction Organization During S. pneumoniae Infection of the Respiratory Epithelium

Many respiratory pathogens compromise epithelial barrier function during lung infection by disrupting intercellular junctions, such as adherens junctions and tight junctions, that maintain intercellular integrity. This includes Streptococcus pneumoniae, a leading cause of pneumonia, which can successfully breach the epithelial barrier and cause severe infections such as septicemia and meningitis. Fluorescence microscopy analysis on intercellular junction protein manipulation by respiratory pathogens has yielded major advances in our understanding of their pathogenesis. Unfortunately, a lack of automated image analysis tools that can tolerate variability in sample-sample staining has limited the accuracy in evaluating intercellular junction organization quantitatively. We have created an open source, automated Python computer script called “Intercellular Junction Organization Quantification” or IJOQ that can handle a high degree of sample-sample staining variability and robustly measure intercellular junction integrity. In silico validation of IJOQ was successful in analyzing computer generated images containing varying degrees of simulated intercellular junction disruption. Accurate IJOQ analysis was further confirmed using images generated from in vitro and in vivo bacterial infection models. When compared in parallel to a previously published, semi-automated script used to measure intercellular junction organization, IJOQ demonstrated superior analysis for all in vitro and in vivo experiments described herein. These data indicate that IJOQ is an unbiased, easy-to-use tool for fluorescence microscopy analysis and will serve as a valuable, automated resource to rapidly quantify intercellular junction disruption under diverse experimental conditions

Pore-forming activity of S. pneumoniae pneumolysin disrupts the paracellular localization of the epithelial adherens junction protein E-cadherin

Streptococcus pneumoniae, a common cause of community-acquired bacterial pneumonia, can cross the respiratory epithelial barrier to cause lethal septicemia and meningitis. S. pneumoniae pore-forming toxin pneumolysin (PLY) triggers robust neutrophil (PMN) infiltration that promotes bacterial transepithelial migration in vitro and disseminated disease in mice. Apical infection of polarized respiratory epithelial monolayers by S. pneumoniae at a multiplicity of infection (MOI) of 20 resulted in recruitment of PMNs, loss of 50% of the monolayer, and PMN-dependent bacterial translocation. Reducing the MOI to 2 decreased PMN recruitment two-fold and preserved the monolayer, but apical-to-basolateral translocation of S. pneumoniae remained relatively efficient. At both MOI of 2 and 20, PLY was required for maximal PMN recruitment and bacterial translocation. Co-infection by wild-type S. pneumoniae restored translocation by a PLY-deficient mutant, indicating that PLY can act in trans. Investigating the contribution of S. pneumoniae infection on apical junction complexes in the absence of PMN transmigration, we found that S. pneumoniae infection triggered the cleavage and mislocalization of the adherens junction (AJ) protein E-cadherin. This disruption was PLY-dependent at MOI of 2 and was recapitulated by purified PLY, requiring its pore-forming activity. In contrast, at MOI of 20, E-cadherin disruption was independent of PLY, indicating that S. pneumoniae encodes multiple means to disrupt epithelial integrity. This disruption was insufficient to promote bacterial translocation in the absence of PMNs. Thus, S. pneumoniae triggers cleavage and mislocalization of E-cadherin through PLY-dependent and -independent mechanisms, but maximal bacterial translocation across epithelial monolayers requires PLY-dependent neutrophil transmigration

A multi-detector array for high energy nuclear e+e- pair spectrosocopy

A multi-detector array has been constructed for the simultaneous measurement of energy- and angular correlation of electron-positron pairs produced in internal pair conversion (IPC) of nuclear transitions up to 18 MeV. The response functions of the individual detectors have been measured with mono-energetic beams of electrons. Experimental results obtained with 1.6 MeV protons on targets containing $^{11}$B and $^{19}$F show clear IPC over a wide angular range. A comparison with GEANT simulations demonstrates that angular correlations of $e^+e^-$ pairs of transitions in the energy range between 6 and 18 MeV can be determined with sufficient resolution and efficiency to search for deviations from IPC due to the creation and subsequent decay into $e^+e^-$ of a hypothetical short-lived neutral boson.Comment: 20 pages, 8 figure

Search for the radiative decay η→π0γγ\eta \to \pi^0 \gamma \gamma in the SND experiment at VEPP-2M

The $\eta \to \pi^0 \gamma \gamma$ decay was investigated by the SND detector at VEPP-2M $e^+e^-$ collider in the reaction $e^+e^-\to\phi\to \eta\gamma$. Here we present the results and some details of this study. We report an upper limit (90% c.l.) $Br(\eta \to \pi^0 \gamma \gamma)<8.4\times 10^{-4}$ as our final result. Our upper limit does not contradict the earlier measurement by GAMS spectrometer. To facilitate future studies a rather detailed review of the problem is also given.Comment: 24 pages, 6 figures, LaTex. To be published in Nucl. Phys.

The antinucleon-nucleon interaction at low energy : annihilation dynamics

The general properties of antiproton-proton annihilation at rest are presented, with special focus on the two-meson final states. The data exhibit remarkable dynamical selection rules : some allowed annihilation modes are suppressed by one order of magnitude with respect to modes of comparable phase-space. Various phenomenological analyses are reviewed, based on microscopic quark dynamics or symmetry considerations. The role of initial- and final-state interaction is also examined.Comment: 128 pages, 49 tables, 27 figure

Single Stage Transoral Laser Microsurgery for Early Glottic Cancer

Objectives: The purpose of the study was to present the outcome of our management protocol of a single stage transoral laser microsurgery (SSTLM), with the intention of complete removal of a lesion, considered to be an early glottic cancer.Methods: Between January 2015 to February 2017 patients with the clinical appearance of an early glottic cancer, who were candidates for (SSTLM) management protocol, were included in this study. Type of cordectomy was determined by pre- and intra-operative evaluation of the extent of lesion in cord layers.Results: Thirty patients (6 females, 24 males; mean age 65 years) underwent SSTLM. Twenty-two patients had malignant histopathological diagnosis of severe dysplasia or Cis in 4 patients, microinvasice carcinoma in 3 patients and invasive carcinoma in 15 patients (T1a tumor in 14 and T1b tumor in 1). Eight patients had a nonmalignant histological diagnosis of keratosis without atypia in 2 patients, mild dysplasia in 2 patients and moderate dysplasia in 3 patients. Based on pre- and intra-operative evaluation, 14 subepithelial (type I), 10 subligamental (type II), and 6 transmuscular (type III) cordectomies were performed. Comparison of cordectomies types with postoperative histopathologic diagnosis showed an adequate extent of resection in 26 out of 30 patients (87%). Considering only patients without recent background of direct laryngoscopy and biopsy, an adequate resection was performed in 90% of patients. None of the patients was further treated by external beam radiation. At average follow-up of 21 months, none of the patients developed local recurrence.Conclusion: In selected cases, a SSTLM for clinical appearance of an early glottic cancer, allows a reliable histopathologic diagnosis and a high local control rate with favorable cost effectiveness. A careful pre- and intraoperative evaluation for selecting the appropriate cases for this management is required in order to avoid under- or over-treatment

Allergic diseases in the elderly

Demographic distribution of the population is progressively changing with the proportion of elderly persons increasing in most societies. This entails that there is a need to evaluate the impact of common diseases, such as asthma and other allergic conditions, in this age segment. Frailty, comorbidities and polymedication are some of the factors that condition management in geriatric patients. The objective of this review is to highlight the characteristics of allergic diseases in older age groups, from the influence of immunosenescence, to particular clinical implications and management issues, such as drug interactions or age-related side effects