Basic science232. Certolizumab pegol prevents pro-inflammatory alterations in endothelial cell function

Background: Cardiovascular disease is a major comorbidity of rheumatoid arthritis (RA) and a leading cause of death. Chronic systemic inflammation involving tumour necrosis factor alpha (TNF) could contribute to endothelial activation and atherogenesis. A number of anti-TNF therapies are in current use for the treatment of RA, including certolizumab pegol (CZP), (Cimzia ®; UCB, Belgium). Anti-TNF therapy has been associated with reduced clinical cardiovascular disease risk and ameliorated vascular function in RA patients. However, the specific effects of TNF inhibitors on endothelial cell function are largely unknown. Our aim was to investigate the mechanisms underpinning CZP effects on TNF-activated human endothelial cells. Methods: Human aortic endothelial cells (HAoECs) were cultured in vitro and exposed to a) TNF alone, b) TNF plus CZP, or c) neither agent. Microarray analysis was used to examine the transcriptional profile of cells treated for 6 hrs and quantitative polymerase chain reaction (qPCR) analysed gene expression at 1, 3, 6 and 24 hrs. NF-κB localization and IκB degradation were investigated using immunocytochemistry, high content analysis and western blotting. Flow cytometry was conducted to detect microparticle release from HAoECs. Results: Transcriptional profiling revealed that while TNF alone had strong effects on endothelial gene expression, TNF and CZP in combination produced a global gene expression pattern similar to untreated control. The two most highly up-regulated genes in response to TNF treatment were adhesion molecules E-selectin and VCAM-1 (q 0.2 compared to control; p > 0.05 compared to TNF alone). The NF-κB pathway was confirmed as a downstream target of TNF-induced HAoEC activation, via nuclear translocation of NF-κB and degradation of IκB, effects which were abolished by treatment with CZP. In addition, flow cytometry detected an increased production of endothelial microparticles in TNF-activated HAoECs, which was prevented by treatment with CZP. Conclusions: We have found at a cellular level that a clinically available TNF inhibitor, CZP reduces the expression of adhesion molecule expression, and prevents TNF-induced activation of the NF-κB pathway. Furthermore, CZP prevents the production of microparticles by activated endothelial cells. This could be central to the prevention of inflammatory environments underlying these conditions and measurement of microparticles has potential as a novel prognostic marker for future cardiovascular events in this patient group. Disclosure statement: Y.A. received a research grant from UCB. I.B. received a research grant from UCB. S.H. received a research grant from UCB. All other authors have declared no conflicts of interes

Comparative evaluation of hemodynamic, vasoconstrictive, and SpO2variability during different stages of periodontal surgery performed using 0.5% ropivacaine or 2% lignocaine HCl (1:80,000 adrenaline) local anesthesia: A randomized, double-blind, split-mouth pilot study

Aim: The aim of this study is to compare anesthetic, hemodynamic, vasoconstrictive, and SpO2variability of 0.5% ropivacaine to the “gold standard” lignocaine (2%) with epinephrine (1:80,000) during periodontal surgery. Materials and Methods: A total of 20 systemically healthy controls meeting the inclusion criteria were selected from the Outpatient Department of Sri Sai College of Dental Surgery. Preoperatively, all participants were infiltrated with 0.5 ml of 0.5% ropivacaine intradermally as test solution to record any allergic reaction. Open flap debridement was performed using local anesthesia containing 2% lignocaine hydrochloride with 1:80,000 epinephrine or 0.5% ropivacaine. Recordings were made of the time of onset, duration of action, the intensity, and depth of anesthesia and various hemodynamic changes throughout the surgical procedure. In addition, blood loss volume and postoperative pain were also assessed. Results: Ropivacaine showed statistically longer duration of action (mean±SD =5.3±0.71 hrs) than lignocaine with epinephrine (mean=2.14±0.98 hrs). Blood loss during flap surgery was comparatively less when performed under ropivacaine. No statistical differences were observed in systolic BP, diastolic BP, SpO2 and heart rate during different stages of periodontal surgery between either of the local anesthetic agents Conclusion: Ropivacaine demonstrates comparable efficacy as lignocaine with added advantage of longer duration of action and superior postoperative pain control. No adverse events from this newer anesthetic were noted, and hence, it can be used safely as a viable local anesthetic for periodontal surgical procedures

Periodontal therapy in orthodontics- A case series

Interdisciplinary approach helps the specialists of contributing disciplines immensely in diagnosis, treatment planning, execution of planned treatment and problem solving for any untoward complication. Orthodontic treatment may be adjunctive to periodontal therapy or vice versa. Periodontal problems might lead to occlusal abnormalities which may require orthodontic correction. Also orthodontic treatment of adult patients is most frequently just one component of a more complex treatment involving several dental disciplines. Various orthodontic treatments also may precipitate some periodontal problems which frequently require periodontal management. This report discusses, through representative cases, potential periodontal problems and their management encountered before, during and after orthodontic treatment

A loop-counting method for covariate-corrected low-rank biclustering of gene-expression and genome-wide association study data

<div><p>A common goal in data-analysis is to sift through a large data-matrix and detect any significant submatrices (i.e., biclusters) that have a low numerical rank. We present a simple algorithm for tackling this biclustering problem. Our algorithm accumulates information about 2-by-2 submatrices (i.e., ‘loops’) within the data-matrix, and focuses on rows and columns of the data-matrix that participate in an abundance of low-rank loops. We demonstrate, through analysis and numerical-experiments, that this loop-counting method performs well in a variety of scenarios, outperforming simple spectral methods in many situations of interest. Another important feature of our method is that it can easily be modified to account for aspects of experimental design which commonly arise in practice. For example, our algorithm can be modified to correct for controls, categorical- and continuous-covariates, as well as sparsity within the data. We demonstrate these practical features with two examples; the first drawn from gene-expression analysis and the second drawn from a much larger genome-wide-association-study (GWAS).</p></div

Illustration of the GSE48091 gene-expression data-set used in Example-A (see main text).

<p>Each row corresponds to a patient, and each column to a ‘gene’ (i.e., gene-expression measurement): the color of each pixel codes for the intensity of a particular measurement of a particular patient (see colorbar to the bottom).<i>M</i>_<i>D</i> = 340 of these patients are cases, the other <i>M</i>_<i>X</i> = 166 are controls; we group the former into the case-matrix ‘<i>D</i>’, and the latter into the control-matrix ‘<i>X</i>’.</p

Illustration of the loops within a 3-dimensional array.

<p>We sketch the structure of a 3-dimensional data-array <i>D</i>, with <i>J</i> rows, <i>K</i> columns and <i>P</i> ‘layers’. Each entry <i>D</i>_{<i>j</i>,<i>k</i>,<i>l</i>} will lie in the cube shown. The loops within <i>D</i> can be divided into 3-categories: (a) iso-layer loops that stretch across 2 rows and 2 columns, (b) iso-column loops that stretch across 2 rows and 2 layers, and (c) iso-row loops that stretch across 2 columns and 2 layers. The row-score [<i>Z</i>_ROW]_<i>j</i> aggregates all the iso-column and iso-layer loops associated with row-<i>j</i>. The column-score [<i>Z</i>_COL]_<i>k</i> aggregates all the iso-row and iso-layer loops associated with column-<i>k</i>. The layer-score [<i>Z</i>_LYR]_<i>l</i> aggregates all the iso-row and iso-column loops associated with layer-<i>l</i>.</p

Contrasting a bicluster with controls.

<p>This shows the bicluster of <a href="http://www.ploscompbiol.org/article/info:doi/10.1371/journal.pcbi.1006105#pcbi.1006105.g005" target="_blank">Fig 5B</a> on top, and the rest of the controls on the bottom. The control-patients have been rearranged in order of their correlation with the co-expression pattern of the bicluster. Even though a few of the controls (i.e,. ∼ 3/166) exhibit a coexpression pattern comparable to that expressed by the bicluster, the vast majority do not.</p